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Sample records for western assays comparison

  1. COMPARISONS OF ELISA AND WESTERN BLOT ASSAYS FOR DETECTION OF CRYPTOSPORIDIUM ANTIBODY

    Science.gov (United States)

    A seroprevalence survey was conducted using ELISA and Western blot (WB) assays for antibody to three Cryptosporidium antigens on 380 blood donors in Jackson County, Oregon. The purpose was to determine if either assay could detect serological evidence of an outbreak which occurre...

  2. Comparisons of ELISA and Western blot assays for detection of autophagy flux.

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    Oh, Sung-Hee; Choi, Yong-Bok; Kim, June-Hyun; Weihl, Conrad C; Ju, Jeong-Sun

    2017-08-01

    We analyzed autophagy/mitophagy flux in vitro (C2C12 myotubes) and in vivo (mouse skeletal muscle) following the treatments of autophagy inducers (starvation, rapamycin) and a mitophagy inducer (carbonyl cyanide m-chlorophenylhydrazone, CCCP) using two immunodetection methods, ELISA and Western blotting, and compared their working range, accuracy, and reliability. The ELISAs showed a broader working range than that of the LC3 Western blots (Table 1). Table 2 showed that data value distribution was tighter and the average standard error from the ELISA was much smaller than those of the Western blot, directly relating to the accuracy of the assay. Test-retest reliability analysis showed good reliability for three individual ELISAs (interclass correlation, ≥ 0.7), but poor reliability for three individual Western blots (interclass correlation, ≤ 0.4) (Table 3).

  3. Quantification of rapid Myosin regulatory light chain phosphorylation using high-throughput in-cell Western assays: comparison to Western immunoblots.

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    Hector N Aguilar

    Full Text Available BACKGROUND: Quantification of phospho-proteins (PPs is crucial when studying cellular signaling pathways. Western immunoblotting (WB is commonly used for the measurement of relative levels of signaling intermediates in experimental samples. However, WB is in general a labour-intensive and low-throughput technique. Because of variability in protein yield and phospho-signal preservation during protein harvesting, and potential loss of antigen during protein transfer, WB provides only semi-quantitative data. By comparison, the "in-cell western" (ICW technique has high-throughput capacity and requires less extensive sample preparation. Thus, we compared the ICW technique to WB for measuring phosphorylated myosin regulatory light chain (PMLC(20 in primary cultures of uterine myocytes to assess their relative specificity, sensitivity, precision, and quantification of biologically relevant responses. METHODOLOGY/PRINCIPAL FINDINGS: ICWs are cell-based microplate assays for quantification of protein targets in their cellular context. ICWs utilize a two-channel infrared (IR scanner (Odyssey(R to quantify signals arising from near-infrared (NIR fluorophores conjugated to secondary antibodies. One channel is dedicated to measuring the protein of interest and the second is used for data normalization of the signal in each well of the microplate. Using uterine myocytes, we assessed oxytocin (OT-stimulated MLC(20 phosphorylation measured by ICW and WB, both using NIR fluorescence. ICW and WB data were comparable regarding signal linearity, signal specificity, and time course of phosphorylation response to OT. CONCLUSION/SIGNIFICANCE: ICW and WB yield comparable biological data. The advantages of ICW over WB are its high-throughput capacity, improved precision, and reduced sample preparation requirements. ICW might provide better sensitivity and precision with low-quantity samples or for protocols requiring large numbers of samples. These features make the ICW

  4. Expanding the available assays: adapting and validating In-Cell Westerns in microfluidic devices for cell-based assays.

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    Paguirigan, Amy L; Puccinelli, John P; Su, Xiaojing; Beebe, David J

    2010-10-01

    Microfluidic methods for cellular studies can significantly reduce costs due to reduced reagent and biological specimen requirements compared with many traditional culture techniques. However, current types of readouts are limited and this lack of suitable readouts for microfluidic cultures has significantly hindered the application of microfluidics for cell-based assays. The In-Cell Western (ICW) technique uses quantitative immunocytochemistry and a laser scanner to provide an in situ measure of protein quantities in cells grown in microfluidic channels of arbitrary geometries. The use of ICWs in microfluidic channels was validated by a detailed comparison with current macroscale methods and shown to have excellent correlation. Transforming growth factor-β-induced epithelial-to-mesenchymal transition of an epithelial cell line was used as an example for further validation of the technique as a readout for soluble-factor-based assays performed in high-throughput microfluidic channels. The use of passive pumping for sample delivery and laser scanning for analysis opens the door to high-throughput quantitative microfluidic cell-based assays that integrate seamlessly with existing high-throughput infrastructure.

  5. Comparison of T24H-his, GST-T24H and GST-Ts8B2 recombinant antigens in western blot, ELISA and multiplex bead-based assay for diagnosis of neurocysticercosis.

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    Hernández-González, Ana; Noh, John; Perteguer, María Jesús; Gárate, Teresa; Handali, Sukwan

    2017-05-15

    Currently, the reference standard assay for the serodiagnosis of neurocysticercosis (NCC) is the lentil lectin-bound glycoproteins/enzyme-linked immunoelectrotransfer blot (LLGP-EITB). The main disadvantage of this technique is the complexity of obtaining and purifying the LLGP extract. This could be solved by replacement with highly specific recombinant antigens from Taenia solium. Based on previous studies, we selected and produced the recombinant Ts8B2 and T24H proteins and applied them to three diagnostic techniques: western blot (WB), enzyme-linked immunosorbent assay (ELISA) and the multiplex bead-based assay (MBA). The Ts8B2 and T24H cDNA sequences were expressed in a prokaryotic system and the corresponding expression products purified; three recombinant proteins were further characterized: T24H-his, GST-T24H and GST-Ts8B2. The proteins on WB, ELISA and MBA were tested against 149 sera from patients with NCC confirmed by brain imaging, 40 sera from patients with other parasitic diseases, and 131 sera from US. individuals without evidence of neurocysticercosis (clinical/serological/brain imaging). The sensitivity and specificity of each antigen by WB were calculated by counting the number of true positive, false positive, true negative and false negative results. Using the receiver operating characteristic (ROC) curves, the cut-off values for the ELISA and MBA were established as well as the sensitivity and specificity of each assay. All three antigens showed a high sensitivity on WB in active NCC cases with two or more viable cysts and low sensitivity for cases with single viable cyst or calcified lesions and inactive NCC. WB showed the highest specificity and sensitivity out of the three diagnostic techniques. The recombinant T24H-his was the best diagnostic reagent in WB (100% sensitivity, 99.4% specificity), exhibiting similar results to the LLGP-EITB, against the same panel of NCC sera. The GST-T24H antigen worked better than the others in ELISA and MBA

  6. Multicentre comparison of a diagnostic assay

    DEFF Research Database (Denmark)

    Waters, Patrick; Reindl, Markus; Saiz, Albert;

    2016-01-01

    ) assays in neuromyelitis optica spectrum disorders (NMOSD). METHODS: Coded samples from patients with neuromyelitis optica (NMO) or NMOSD (101) and controls (92) were tested at 15 European diagnostic centres using 21 assays including live (n=3) or fixed cell-based assays (n=10), flow cytometry (n=4...

  7. A Comparison between Chinese and Western Culture

    Institute of Scientific and Technical Information of China (English)

    HE Ting-ting

    2015-01-01

    Different countries have different cultures. Understanding the differences between Chinese and western culture will help us to communicate with each other better. From three aspects, this paper mainly compares the differences between Chinese and western culture.

  8. The line blot assay: problems with titrating first and second antibodies for Western blot and immunohistochemistry assays?

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    Rojas-Espinosa, O; Silva-Miranda, M; Wek-Rodriguez, K; Arce-Paredes, P

    2006-01-01

    We describe a technique designed to assess the optimal dilution of primary and secondary antibodies, to be used in Western blot, dot blot, the multi-antigen print immunoassay (MAPIA) and immunohistochemistry assays. The method that we call "line blot" is not an alternative but a practical, complementary tool for the above techniques that assures definitive results are obtained from single assays, so there is no need to repeat the assay. As with most immunoenzymatic assays, the line blot assay is very sensitive, allowing the detection of absolute amounts of antigen as low as 2.5 ng in the 0.5 cm-long segment line (see Results), depending on the strength of the secondary, enzyme-labelled antibody.

  9. Validation of western blot for Histoplasma capsulatum antibody detection assay.

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    Almeida, Marcos de Abreu; Pizzini, Cláudia Vera; Damasceno, Lisandra Serra; Muniz, Mauro de Medeiros; Almeida-Paes, Rodrigo; Peralta, Regina Helena Saramago; Peralta, José Mauro; Oliveira, Raquel de Vasconcelos Carvalhaes; Vizzoni, Alexandre Gomes; de Andrade, Carla Lourenço Tavares; Zancopé-Oliveira, Rosely Maria

    2016-02-24

    Histoplasmosis is worldwide systemic mycoses caused by the dimorphic fungus Histoplasma capsulatum. The isolation and identification of H. capsulatum in culture is the reference test for histoplasmosis diagnosis confirmation. However, in the absence of it, serology has been used as a presumptive diagnosis through antibody and antigen detection. The purpose of the present study was to validate an immunoassay method (western blot) for antibodies detection in the diagnosis of histoplasmosis. To validate the western blot (WB) a study was conducted using 118 serum samples from patients with histoplasmosis and 118 serum controls collected from January 2000 to December 2013 in residents of the Rio de Janeiro State, Brazil. Diagnostic validation parameters were calculated based on the categorization of results obtained in a 2 × 2 table and subjected to statistical analysis. In addition, the viability of deglycosylated histoplasmin antigen (ptHMIN) onto nitrocellulose membranes previously sensitized was evaluated during the same period. The WB test showed sensitivity of 94.9 %, specificity of 94.1 %, positive predictive value of 94.1 %, negative predictive value of 94.9 %, accuracy of 94.5 %, and almost perfect precision. Besides, the strips have proved to be viable for using at least 5 years after ptHMIN antigen sensitization. Western blot test using ptHMIN provides sensitive, specific, and faster results. Therefore, could be considered a useful tool in the diagnosis of histoplasmosis being used by public health system, even in situations where laboratory facilities are relatively limited.

  10. Assay-dependent variability of serum insulin concentrations: a comparison of eight assays.

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    Tohidi, Maryam; Arbab, Parvaneh; Ghasemi, Asghar

    2017-04-01

    Although insulin measurement is essential for both clinical and research purposes, there is currently no reference method for insulin assays. The aim of this study was to compare results of serum insulin determined by a number of commercially available assays. We compared eight insulin assays by analyzing 165 serum samples. Assays included two chemiluminescence (Roche and DiaSorin), four ELISA (Tosoh, Mercodia, Monobind, and Diametra), and two IRMA (Izotop and BioSource) methods. Each assay was compared with the mean of all assay methods and Bland-Altman difference plots were used to measure agreement between each assay and overall mean. Least squared perpendicular distance regression analysis (Deming's method) was used to calculate slope and intercept for bias and also for each assay vs. mean of eight assays. Findings showed that the lowest and highest median insulin concentrations varied by a factor of 1.8. Maximum and minimum correlations with mean of assays were observed for Roche (0.992) and BioSource (0.844), respectively. Significant bias was observed in six assays. In pairwise comparisons of different assays, the highest and least mean differences were 7.78 μU/mL and -0.14 μU/mL, respectively. In conclusion, serum insulin measurement with different assays showed a maximum of 1.8-fold difference, a point that should be taken into consideration in the interpretation of circulating insulin levels in both clinical and research fields.

  11. Comparison of immunoturbidimetric and immunonephelometric assays for specific proteins.

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    Mali, Bahera; Armbruster, David; Serediak, Ernie; Ottenbreit, Tammy

    2009-10-01

    Immunoturbidimetric assays for specific proteins are available on "open system" clinical chemistry analyzers. The analytical performance of nine immunoturbidimetric specific protein assays (C3, C4, CRP, Haptoglobin, IgA, IgG, IgM, RF, and Transferrin) was compared to immunonephelometry. Testing was performed on the Abbott ARCHITECT ci8200 and the Dade Behring BNII nephelometer and evaluated for precision, linearity, limit of detection, prozone phenomenon, method comparison, workflow, and proficiency testing survey comparison. Immunoturbidimetric assays performance was satisfactory for total precision, linearity, limit of detection and the prozone effect was not observed. Method comparison was acceptable for the immunoglobulins, CRP and transferrin but less favorable for the other assays, likely due to methodology and antibody specificity differences. Immunourbidimetric specific protein assays allow for efficient test consolidation on a general purpose clinical chemistry analyzer.

  12. Comparison of bacterial culture, polymerase chain reaction, and a mix-enzyme-linked immunosorbent assay for the detection of Salmonella status in grow-to-finish pigs in western Canada with a Bayesian approach

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    Wilkins, Wendy; Waldner, Cheryl; Rajić, Andrijana; McFall, Margaret; Chow, Eva; Muckle, Anne

    2011-01-01

    Among grow-to-finish pigs from 10 herds in Alberta and Saskatchewan, 23 (16%) of 144 fecal samples were culture-positive and 40 (28%) of 144 pigs were seropositive for Salmonella. With a Bayesian model specifying dependence between the 2 tests, the sensitivity (Se) of culture and real-time polymerase chain reaction (RT-PCR) was 79% to 86%, depending on the cut-off value for the enzyme-linked immunosorbent assay (ELISA). Culture specificity (Sp) was assumed to be 100%; RT-PCR Sp was found to be 94%. The ELISA Se was 76% and 51% at optical density cut-off values ≥ 20% and ≥ 40%, respectively; the Sp was 94% at each cut-off value. The model showed some sensitivity to ELISA prior information, the ELISA Se being approximately 8% lower when informative prior information was specified in the model. When there was no adjustment for dependence between culture and RT-PCR, the posterior estimates for both culture and RT-PCR Se were 11% higher than with the conditional-dependence model and had considerably narrower probability intervals, which suggests that correlation between culture and PCR is important and should be adjusted for in future studies. PMID:22468029

  13. Imported intraocular gnathostomiasis with subretinal tracks confirmed by western blot assay.

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    Yang, Ji Ho; Kim, Moosang; Kim, Eung Suk; Na, Byoung-Kuk; Yu, Seung-Young; Kwak, Hyung-Woo

    2012-03-01

    We report a case of intraocular gnathostomiasis diagnosed by western blot assay in a patient with subretinal tracks. A 15-year-old male patient complained of blurred vision in the right eye, lasting for 2 weeks. Eight months earlier, he had traveled to Vietnam for 1 week and ate raw wild boar meat and lobster. His best-corrected visual acuity was 20/20 in both eyes and anterior chamber examination revealed no abnormalities. Fundus examination showed subretinal tracks in the right eye. Fluorescein angiography and indocyanine green angiography showed linear hyperfluorescence of the subretinal lesion observed on fundus in the right eye. Ultrasound examination revealed no abnormalities. Blood tests indicated mild eosinophilia (7.5%), and there was no abnormality found by systemic examinations. Two years later, the patient visited our department again for ophthalmologic evaluation. Visual acuity remained 20/20 in both eyes and the subretinal tracks in the right eye had not changed since the previous examination. Serologic examination was performed to provide a more accurate diagnosis, and the patient's serum reacted strongly to the Gnathostoma nipponicum antigen by western blot assay, which led to a diagnosis of intraocular gnathostomiasis. This is the first reported case of intraocular gnathostomiasis with subretinal tracks confirmed serologically using western blot in Korea.

  14. Determining vitamin D status: a comparison between commercially available assays.

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    Greta Snellman

    Full Text Available BACKGROUND: Vitamin D is not only important for bone health but can also affect the development of several non-bone diseases. The definition of vitamin D insufficiency by serum levels of 25-hydroxyvitamin D depends on the clinical outcome but might also be a consequence of analytical methods used for the definition. Although numerous 25-hydroxyvitamin D assays are available, their comparability is uncertain. We therefore aim to investigate the precision, accuracy and clinical consequences of differences in performance between three common commercially available assays. METHODOLOGY/PRINCIPAL FINDINGS: Serum 25-hydroxyvitamin D levels from 204 twins from the Swedish Twin Registry were determined with high-pressure liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry (HPLC-APCI-MS, a radioimmunoassay (RIA and a chemiluminescent immunoassay (CLIA. High inter-assay disagreement was found. Mean 25-hydroxyvitamin D levels were highest for the HPLC-APCI-MS technique (85 nmol/L, 95% CI 81-89, intermediate for RIA (70 nmol/L, 95% CI 66-74 and lowest with CLIA (60 nmol/L, 95% CI 56-64. Using a 50-nmol/L cut-off, 8% of the subjects were insufficient using HPLC-APCI-MS, 22% with RIA and 43% by CLIA. Because of the heritable component of 25-hydroxyvitamin D status, the accuracy of each method could indirectly be assessed by comparison of within-twin pair correlations. The strongest correlation was found for HPLC-APCI-MS (r = 0.7, intermediate for RIA (r = 0.5 and lowest for CLIA (r = 0.4. Regression analyses between the methods revealed a non-uniform variance (p<0.0001 depending on level of 25-hydroxyvitamin D. CONCLUSIONS/SIGNIFICANCE: There are substantial inter-assay differences in performance. The most valid method was HPLC-APCI-MS. Calibration between 25-hydroxyvitamin D assays is intricate.

  15. Western blot assay for quantitative and qualitative antigen detection in vaccine development.

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    Kumar, Sanjai; Zheng, Hong; Mahajan, Babita; Kozakai, Yukiko; Morin, Merribeth; Locke, Emily

    2014-05-01

    Immunological methods for quantitative measurement, antigenic characterization, and monitoring the stability of active immunogenic component(s) are a critical need in the vaccine development process. This unit describes an enhanced chemiluminescence-based western blot for quantitative detection of Plasmodium falciparum circumsporozoite protein (PfCSP), a major malaria candidate vaccine antigen. The most salient features of this assay are its high sensitivity and reproducibility; it can reliably detect ∼5 to 10 pg PfCSP expressed on native parasites or recombinantly expressed in Escherichia coli. Although described for a specific vaccine antigen, this assay should be applicable for any antigen-antibody combination for which relevant detection reagents are available. Detailed stepwise experimental procedures and methods for data acquisition and analysis are described. Copyright © 2014 John Wiley & Sons, Inc.

  16. A new Western blot assay for the detection of porcine cytomegalovirus (PCMV).

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    Plotzki, Elena; Keller, Martina; Ivanusic, Daniel; Denner, Joachim

    2016-10-01

    Porcine cytomegalovirus (PCMV) may be harmful for human recipients if xenotransplantation using pig cell, tissue or organ will be performed transmitting the virus from donor pigs to human recipients. PCMV is widespread in pigs and closely related to human pathogenic herpesviruses, however there are no data concerning infection of humans. In contrast, recently it had been shown that transplantation of organs from pigs infected with PCMV into non-human primate recipients resulted in a significant reduction of the survival time compared with the transplantation of organs from uninfected pigs. To prevent transmission of PCMV in future pig to human xenotransplantations, sensitive and specific detection methods should be used. Here a new Western blot assay using recombinant proteins corresponding to two domains of the glycoprotein gB of PCMV is described. With this assay, the presence of PCMV-specific antibodies in different pig breeds was analysed. Antibodies were detected in a high percentage of animals, in one breed up to 85%. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Comparison and standardization of soil enzyme assay for meaningful data interpretation.

    Science.gov (United States)

    Deng, Shiping; Dick, Richard; Freeman, Christopher; Kandeler, Ellen; Weintraub, Michael N

    2017-02-01

    Data interpretation and comparison in enzyme assays can be challenging because of the complex nature of the environment and variations in methods employed. This letter provides an overview of common enzyme assays, the need for methods standardization, and solutions addressing some of the concerns in microplate fluorimetric assay approaches.

  18. Comparison of five assays for detection of Clostridium difficile toxin.

    Science.gov (United States)

    Chapin, Kimberle C; Dickenson, Roberta A; Wu, Fongman; Andrea, Sarah B

    2011-07-01

    Performance characteristics of five assays for detection of Clostridium difficile toxin were compared using fresh stool samples from patients with C. difficile infection (CDI). Assays were performed simultaneously and according to the manufacturers' instructions. Patients were included in the study if they exhibited clinical symptoms consistent with CDI. Nonmolecular assays included glutamate dehydrogenase antigen tests, with positive findings followed by the Premier Toxin A and B Enzyme Immunoassay (GDH/EIA), and the C. Diff Quik Chek Complete test. Molecular assays (PCR) included the BD GeneOhm Cdiff Assay, the Xpert C. difficile test, and the ProGastro Cd assay. Specimens were considered true positive if results were positive in two or more assays. For each method, the Youden index was calculated and cost-effectiveness was analyzed. Of 81 patients evaluated, 26 (32.1%) were positive for CDI. Sensitivity of the BD GeneOhm Cdiff assay, the Xpert C. difficile test, the ProGastro Cd assay, C. Diff Quik Chek Complete test, and two-step GDH/EIA was 96.2%, 96.2%, 88.5%, 61.5%, and 42.3%, respectively. Specificity of the Xpert C. difficile test was 96.4%, and for the other four assays was 100%. Compared with nonmolecular methods, molecular methods detected 34.7% more positive specimens. Assessment of performance characteristics and cost-effectiveness demonstrated that the BD GeneOhm Cdiff assay yielded the best results. While costly, the Xpert C. difficile test required limited processing and yielded rapid results. Because of discordant results, specimen processing, and extraction equipment requirements, the ProGastro Cd assay was the least favored molecular assay. The GDH/EIA method lacked sufficient sensitivity to be recommended. Copyright © 2011 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.

  19. Evaluation of urine CCA assays for detection of Schistosoma mansoni infection in Western Kenya.

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    Hillary L Shane

    Full Text Available Although accurate assessment of the prevalence of Schistosoma mansoni is important for the design and evaluation of control programs, the most widely used tools for diagnosis are limited by suboptimal sensitivity, slow turn-around-time, or inability to distinguish current from former infections. Recently, two tests that detect circulating cathodic antigen (CCA in urine of patients with schistosomiasis became commercially available. As part of a larger study on schistosomiasis prevalence in young children, we evaluated the performance and diagnostic accuracy of these tests--the carbon test strip designed for use in the laboratory and the cassette format test intended for field use. In comparison to 6 Kato-Katz exams, the carbon and cassette CCA tests had sensitivities of 88.4% and 94.2% and specificities of 70.9% and 59.4%, respectively. However, because of the known limitations of the Kato-Katz assay, we also utilized latent class analysis (LCA incorporating the CCA, Kato-Katz, and schistosome-specific antibody results to determine their sensitivities and specificities. The laboratory-based CCA test had a sensitivity of 91.7% and a specificity of 89.4% by LCA while the cassette test had a sensitivity of 96.3% and a specificity of 74.7%. The intensity of the reaction in both urine CCA tests reflected stool egg burden and their performance was not affected by the presence of soil transmitted helminth infections. Our results suggest that urine-based assays for CCA may be valuable in screening for S. mansoni infections.

  20. Western blot analysis of adhesive interactions under fluid shear conditions: the blot rolling assay.

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    Sackstein, Robert; Fuhlbrigge, Robert

    2015-01-01

    Western blotting has proven to be an important technique in the analysis of receptor-ligand interactions (i.e., by ligand blotting) and for identifying molecules mediating cell attachment (i.e., by cell blotting). Conventional ligand blotting and cell blotting methods employ non-dynamic (static) incubation conditions, whereby molecules or cells of interest are placed in suspension and overlaid on membranes. However, many cell-cell and cell-matrix adhesive interactions occur under fluid shear conditions, and shear stress itself mediates and/or facilitates the engagement of these physiologically appropriate receptors and ligands. Notably, shear forces critically influence the adhesion of circulating cells and platelets to vessel walls in physiologic cell migration and hemostasis, as well as in inflammatory and thrombotic disorders, cancer metastasis, and atherosclerosis. Use of non-dynamic blotting conditions to analyze such interactions can introduce bias, overtly missing relevant effectors and/or exaggerating the relative role(s) of non-physiologic adhesion molecules. To address this shortfall, we have developed a new technique for identifying binding interactions under fluid shear conditions, the "blot rolling assay." Using this method, molecules in a complex mixture are resolved by gel electrophoresis, transferred to a membrane that is rendered semitransparent, and the membrane is then incorporated into a parallel-plate flow chamber apparatus. Under controlled flow conditions, cells or particles bearing adhesion proteins of interest are then introduced into the chamber and interactions with individual immobilized molecules (bands) can be visualized in real time. The substrate molecule(s) supporting adhesion under fluid shear can then be identified by staining with specific antibodies or by excising the relevant band(s) and performing mass spectrometry or microsequencing of the isolated material. This method thus allows for the identification, within a complex

  1. A novel assay detecting recall response to Mycobacterium tuberculosis: Comparison with existing assays.

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    Hsu, Denise C; Zaunders, John J; Plit, Marshall; Leeman, Craig; Ip, Susanna; Iampornsin, Thatri; Pett, Sarah L; Bailey, Michelle; Amin, Janaki; Ubolyam, Sasiwimol; Avihingsanon, Anchalee; Ananworanich, Jintanat; Ruxrungtham, Kiat; Cooper, David A; Kelleher, Anthony D

    2012-07-01

    A strategy to reduce the burden of active TB is isoniazid preventive therapy for latent TB infection (LTBI). However, current assays used to diagnose LTBI all have limitations. In these proof of concept studies, we compared the agreement of a novel flow cytometry assay detecting CD25/CD134 co-expression with QuantiFERON-TB Gold In-Tube (QFN-GIT) and Tuberculin skin test (TST) in the detection of recall immune response to TB. The CD25/CD134 assay, QFN-GIT and TST were performed on 74 participants referred for TB screening in Sydney and on 50 participants with advanced HIV infection (CD4 ≤ 350 × 10(6) cells/L) in Bangkok. The agreement between CD25/CD134 assay and QFN-GIT was 93.2% (Kappa 0.631 95% CI 0.336-0.926) in Sydney and 90% (Kappa 0.747 95% CI 0.541-0.954) in Bangkok. Discordant results occurred around the cut off of both tests. The agreement between CD25/CD134 assay and TST was 73.6% (Kappa 0.206 95% CI 0.004-0.409) in Sydney and 84% (Kappa 0.551 95% CI 0.296-0.806) in Bangkok. The CD25/CD134 assay showed good agreement with QFN-GIT in detecting recall response to TB both in well and less resourced setting as well as in persons with advanced HIV infection. Further study into the performance of this assay is thus warranted.

  2. Mycoplasma agassizii strain variation and distinct host antibody responses explain differences between enzyme-linked immunosorbent assays and Western blot assays.

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    Wendland, Lori D; Klein, Paul A; Jacobson, Elliott R; Brown, Mary B

    2010-11-01

    The precarious status of desert (Gopherus agassizii) and gopher (G. polyphemus) tortoises has resulted in conservation efforts that now include health assessment as an important component of management decision-making. Mycoplasmal upper respiratory tract disease (URTD) is one of very few diseases in chelonians for which comprehensive and rigorously validated diagnostic tests exist. In this study, serum samples obtained from eight Gopherus tortoises documented at necropsy to (i) be enzyme-linked immunosorbent assay (ELISA) seropositive using the PS6 antigen, (ii) be infected with Mycoplasma agassizii as indicated by direct isolation of the pathogen from the respiratory surfaces, and (iii) have histological lesions of mycoplasmal URTD were used to evaluate four distinct clinical isolates of M. agassizii as antigens for ELISA and Western blot analyses. Each animal sample reacted in the Western blot with its homologous M. agassizii strain, but recognition of heterologous M. agassizii strains was variable. Further, individual animals varied significantly with respect to the specific proteins recognized by the humoral immune response. An additional 114 Gopherus serum samples were evaluated using ELISA antigens prepared from the four distinct M. agassizii strains; A₄₀₅ values were significantly correlated (r² goodness of fit range, 0.708 to 0.771; P Western blot binding patterns. Thus, reliance on a single M. agassizii strain as an antigen in Western blot assays may provide false-negative results. This could have adverse consequences for the well-being of these environmentally sensitive hosts if false-negative animals were relocated to sites consisting of true-negative populations.

  3. Comparison of Several Geoid Models over the Western Mediterranean Sea

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    Termens, A.; Martinez-Benjamin, J. J.

    2011-07-01

    The Mediterranean Sea is a semi-enclosed true ocean. Recent Mediterranean circulation and sea level studies using various observations and ocean general circulation models show good coherence and agreement. The satellite altimetry and tide gauge observed and model predicted sea level show good coherent with correlation coefficient of 0.6. The barotropic pressure response accounts for about 66% of the Mediterranean sea level rise (1948-2001). The estimated sea level trend (1.54 ± 0.75 mm/yr) using decadal altimetry (1985-2001) after correcting the interannual/decadal signals reconstructed using tide gauge data, agrees well with the long term trend (1948-2001) estimated using tide gauges (1.43 ± 0.09 mm/yr) in the Mediterranean Sea, and is in better agreement than before with the global long-term sea level trend (1.7 - 1.8 mm/yr). Simulation studies indicate that the time-varying mass variations of Mediterranean Sea likely are sensitive to GOCE at the few mEötvös level. One of GOCE's primary high-level data products is the global gravity model with anticipated geoid accuracy of 1 cm RMS and a spatial resolution of 130 km or longer. Actually, the International Centre for Global Earth Models (ICGEM) distributes some GOCE's Global Gravity field Models (GGMs) like GO_CONS_GCF_2_DIR (Bruinsma et al, 2010), GO_CONS_GCF_2_TIM (Pail et al, 2010a), GO_CONS_GCF_2_SPW (Migliaccio et al, 2010), GOCO01S (Pail et al, 2010b). The work focuses on the comparison between these GOCE's GGMs, EGM2008 and EIGEN-51C, with sea gravity anomalies and geoid undulations provided by existing local and regional geoids - like IBERGEO (Sevilla, 2008), IGG (Corchete et al, 2005), etc. - in the Western Mediterranean Sea in order to find the GGM that best fits this area. We also try to estimate how the GOCE geoid data, provided by ESA, works on the Western Mediterranean Sea in order to prepare future geomatic issues.

  4. Comparison of Different Promoter Methylation Assays in Breast Cancer

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    Karijn P. M. Suijkerbuijk

    2010-01-01

    Full Text Available Background: Promoter hypermethylation has emerged as a promising cancer biomarker. Currently, a large variety of quantitative and non-quantitative techniques is used to measure methylation in clinical specimens. Here we directly compared three commonly used methylation assays and assessed the influence of tissue fixation, target sequence location and the amount of DNA on their performance.

  5. Comparison of love in Chinese culture and in western culture

    Institute of Scientific and Technical Information of China (English)

    喻远洋

    2009-01-01

    In this passage, love in Chinese culture and in western culture is compared. It briefly introduces the difference love view in western culture and in Chinese culture on a specific matter. Then it discusses the origin of this phenomenon. At last it tries to conclude the characters of these two different love views.

  6. A Brief Comparison of Chinese and Western Thinking Patterns:Synthesis, Analysis, and Antithesis

    Institute of Scientific and Technical Information of China (English)

    谭笑

    2013-01-01

    A brief comparison of Chinese and Western thinking patterns reflects in synthesis and analysis, which including tortuous thinking pattern and straightforward thinking pattern. These two different thinking patterns influence our social life everywhere.

  7. Comparison of destructive and nondestructive assay of heterogeneous salt residues

    Energy Technology Data Exchange (ETDEWEB)

    Fleissner, J.G.; Hume, M.W.

    1986-03-29

    To study problems associated with nondestructive assay (NDA) measurements of molten salt residues, a joint study was conducted by the Rocky Flats Plant, Golden, CO and Mound Laboratories, Miamisburg, OH. Extensive NDA measurements were made on nine containers of molten salt residues by both Rocky Flats and Mound followed by dissolution and solution quantification at Rocky Flats. Results of this study verify that plutonium and americium can be measured in such salt residues by a new gamma-ray spectral analysis technique coupled with calorimetry. Biases with respect to the segmented gamma-scan technique were noted.

  8. COMPARISON OF SAMPLE PREPARATION METHODS FOR CHIP-CHIP ASSAYS

    OpenAIRE

    O'Geen, Henriette; Nicolet, Charles M.; Blahnik, Kim; Green, Roland; Farnham, Peggy J.

    2006-01-01

    A single ChIP sample does not provide enough DNA for hybridization to a genomic tiling array. A commonly used technique for amplifying the DNA obtained from ChIP assays is linker-mediated PCR (LMPCR). However, using this amplification method, we could not identify Oct4 binding sites on genomic tiling arrays representing 1% of the human genome (ENCODE arrays). In contrast, hybridization of a pool of 10 ChIP samples to the arrays produced reproducible binding patterns and low background signals...

  9. Comparison of automated von Willebrand factor activity assays

    DEFF Research Database (Denmark)

    Timm, Annette; Hillarp, Andreas; Philips, Malou

    2015-01-01

    activity/antigen ratios in samples classified as having VWD (activity classification power might interfere with the interpretation......INTRODUCTION: Von Willebrand Disease (VWD) is the most common inherited bleeding disorder. Measurement of von Willebrand factor (VWF) activity in plasma is often based on platelet agglutination stimulated by the ristocetin cofactor activity. Novel assays, based on latex beads with recombinant...... glycoprotein Ib instead of platelets, have recently been developed but it is unclear whether these can improve the diagnostic capability for VWD. AIM: To compare four automated VWF activity methods in a mixed population of patients referred for evaluation of bleeding tendency. METHODS: The analytical...

  10. Comparison of Marriage and Family style between China and Western Countries

    Institute of Scientific and Technical Information of China (English)

    马全海; 卫华

    2008-01-01

    The author gives a brief introduction to the marriage and family styles of China and western countries.and makes some brief comparisons of them.In the comparison of the family styles.we can see the diversity of the culture in different coun tries.To study the cultural difference is of great meaning for language learners,and the more we learn about their culture,the better we learn their language.Learning their family and marriage style can help us a lot in learning the life of westerners and hence help us in the international communication of people from western countries.

  11. Comparison of clonogenic assay with premature chromosome condensation assay in prediction of human cell radiosensitivity

    Institute of Scientific and Technical Information of China (English)

    Zhuan-Zi Wang; Wen-Jian Li; Hong Zhang; Jian-She Yang; Rong Qiu; Xiao Wang

    2006-01-01

    AIM: To determine whether the number of non-rejoining G2-chromatid breaks can predict the radiosensitivity of human cell lines.METHODS: Cell lines of human ovary carcinoma cells (HO8910), human hepatoma cells (HepG2) and liver cells (L02) were irradiated with a range of doses and assessed both of cell survival and non-rejoining G2-chromatid breaks at 24 h after irradiation. Cell survival was documented by a colony assay. Non-rejoining G2-chromatid breaks were measured by counting the number of non-rejoining G2 chromatid breaks at 24 h after irradiation, detected by the prematurely chromosome condensed (PCC) technique.RESULTS: A linear-quadratic survival curve was observed in three cell lines, and HepG2 was the most sensitive to y-radiation. A dose-dependent linear increase was observed in radiation-induced non-rejoining G2-PCC breaks measured at 24 h after irradiation in all cell lines, and HepG2 was the most susceptible to induction of non-rejoining G2-PCC breaks. A close correlation was found between the clonogenic radiosensitivity and the radiation-induced non-rejoining G2-PCC breaks (r= 0.923). Furthermore, survival-aberration correlations for two or more than two doses lever were also significant.CONCLUSION: The number of non-rejoining G2 PCC breaks holds considerable promise for predicting the radiosensitivity of normal and tumor cells when two or more than two doses lever is tested.

  12. Interlaboratory comparison of dicentric chromosome assay using electronically transmitted images.

    Science.gov (United States)

    García, O; Di Giorgio, M; Vallerga, M B; Radl, A; Taja, M R; Seoane, A; De Luca, J; Stuck Oliveira, M; Valdivia, P; Lamadrid, A I; González, J E; Romero, I; Mandina, T; Pantelias, G; Terzoudi, G; Guerrero-Carbajal, C; Arceo Maldonado, C; Espinoza, M; Oliveros, N; Martínez-López, W; Di Tomaso, M V; Méndez-Acuña, L; Puig, R; Roy, L; Barquinero, J F

    2013-04-01

    The bottleneck in data acquisition during biological dosimetry based on a dicentric assay is the need to score dicentrics in a large number of lymphocytes. One way to increase the capacity of a given laboratory is to use the ability of skilled operators from other laboratories. This can be done using image analysis systems and distributing images all around the world. Two exercises were conducted to test the efficiency of such an approach involving 10 laboratories. During the first exercise (E1), the participant laboratories analysed the same images derived from cells exposed to 0.5 and 3 Gy; 100 images were sent to all participants for both doses. Whatever the dose, only about half of the cells were complete with well-spread metaphases suitable for analysis. A coefficient of variation (CV) on the standard deviation of ∼15 % was obtained for both doses. The trueness was better for 3 Gy (0.6 %) than for 0.5 Gy (37.8 %). The number of estimated doses classified as satisfactory according to the z-score was 3 at 0.5 Gy and 8 at 3 Gy for 10 dose estimations. In the second exercise, an emergency situation was tested, each laboratory was required to score a different set of 50 images in 2 d extracted from 500 downloaded images derived from cells exposed to 0.5 Gy. Then the remaining 450 images had to be scored within a week. Using 50 different images, the CV on the estimated doses (79.2 %) was not as good as in E1, probably associated to a lower number of cells analysed (50 vs. 100) or from the fact that laboratories analysed a different set of images. The trueness for the dose was better after scoring 500 cells (22.5 %) than after 50 cells (26.8 %). For the 10 dose estimations, the number of doses classified as satisfactory according to the z-score was 9, for both 50 and 500 cells. Overall, the results obtained support the feasibility of networking using electronically transmitted images. However, before its implementation some issues should be elucidated, such as the

  13. A Comparison of Food in the Chinese and Western Cultures

    Institute of Scientific and Technical Information of China (English)

    钮贵芳

    2005-01-01

    <正>Foods cultures are so deeply rooted in Chinese culture that almost everything of Chinese people’s lives is more or less related to them. In the west, foods also have a very important role in people s lives. However, foods in Chinese culture and western culture are different in a few aspects. This paper aims to look into the similarities and differences of the foods in two cultures and tries to give a brief discussion on their differences.

  14. A Comparison of Taboos between Chinese Culture and Western Culture

    Institute of Scientific and Technical Information of China (English)

    邹艳飞

    2013-01-01

    Taboo is a common phenomenon in Chinese and Western Culture. The different cultural backgrounds lead to the differences of taboos. This paper focuses on the differences of taboos from several aspects, such as name title, religion, numbers, privacy topics, etc. According to the analyses, we’l decrease and avoid the cultural shock in intercultural communication. If we know something about taboos in other cultures, we’l succeed in communicating with other people. Otherwise we’l fail to communicate.

  15. Exploring tool innovation: a comparison of Western and Bushman children.

    Science.gov (United States)

    Nielsen, Mark; Tomaselli, Keyan; Mushin, Ilana; Whiten, Andrew

    2014-10-01

    A capacity for constructing new tools, or using old tools in new ways, to solve novel problems is a core feature of what it means to be human. Yet current evidence suggests that young children are surprisingly poor at innovating tools. However, all studies of tool innovation to date have been conducted with children from comparatively privileged Western backgrounds. This raises questions as to whether or not previously documented tool innovation failure is culturally and economically specific. In the current study, thus, we explored the innovation capacities of children from Westernized urban backgrounds and from remote communities of South African Bushmen. Consistent with past research, we found tool innovation to occur at extremely low rates and that cultural background had no bearing on this. The current study is the first to empirically test tool innovation in children from non-Western backgrounds, with our data being consistent with the view that despite its key role in human evolution, a capacity for innovation in tool making remains remarkably undeveloped during early childhood.

  16. The diagnosis of proventricular dilatation disease: use of a Western blot assay to detect antibodies against avian Borna virus.

    Science.gov (United States)

    Villanueva, Itamar; Gray, Patricia; Mirhosseini, Negin; Payne, Susan; Hoppes, Sharman; Honkavuori, Kirsi S; Briese, Thomas; Turner, Debra; Tizard, Ian

    2010-07-14

    Avian Borna virus (ABV) has recently been shown to be the causal agent of proventricular dilatation disease (PDD) a lethal neurologic disease of captive psittacines and other birds. An immunoblot assay was used to detect the presence of antibodies against avian Borna virus in the serum of affected birds. A lysate from ABV-infected duck embryo fibroblasts served as a source of antigen. The assay was used to test for the presence of antibodies to ABV in 117 birds. Thirty of these birds had biopsy or necropsy-confirmed proventricular dilatation disease (PDD), while the remaining 87 birds were apparently healthy or were suffering from diseases other than PDD. Sera from 27 of the 30 PDD cases (90%) contained antibodies to ABV. Seventy-three (84%) of the apparently "healthy" birds were seronegative. Additionally, sera from seven macaws and one parrot trapped in the Peruvian Amazon were seronegative. Positive sera recognized the bornaviral nucleoprotein (N-protein). While the presence of antibodies to ABV largely corresponded with the development of clinical PDD, 14 apparently healthy normal birds possessed detectable antibodies to ABV. The existence of a carrier state was confirmed when 13 of 15 apparently healthy cockatiels were shown by PCR to have detectable ABV RNA in their feces. Western blot assays may be of significant assistance in diagnosing proventricular dilatation disease. Many apparently healthy birds may however be seronegative while, at the same time, shedding ABV in their feces. (c) 2009 Elsevier B.V. All rights reserved.

  17. Comparison of Wine Culture between China and Western Countries

    Institute of Scientific and Technical Information of China (English)

    杨婷婷

    2014-01-01

    With the accelerating pace of globalization, the cultural differences have more impacts and restrictions on the intercom-munication than before. They exist in the total international communication. Therefore, the critical factors whether we can suc-ceed in intercommunication are whether we can eliminate cultural differences or avoid their conflicts. There is no exception for wine culture. As we know, drinking wine plays an important role in public relations as a communicating tool. This essay is to compare Chinese wine culture with the Western one, and try to analyze the factors aroused by the cultural differences, and then provide the solution how to face it.

  18. International network for comparison of HIV neutralization assays: the NeutNet report II.

    Directory of Open Access Journals (Sweden)

    Leo Heyndrickx

    Full Text Available BACKGROUND: Neutralizing antibodies provide markers for vaccine-induced protective immunity in many viral infections. By analogy, HIV-1 neutralizing antibodies induced by immunization may well predict vaccine effectiveness. Assessment of neutralizing antibodies is therefore of primary importance, but is hampered by the fact that we do not know which assay(s can provide measures of protective immunity. An international collaboration (NeutNet involving 18 different laboratories previously compared different assays using monoclonal antibodies (mAbs and soluble CD4 (Phase I study. METHODS: In the present study (Phase II, polyclonal reagents were evaluated by 13 laboratories. Each laboratory evaluated nine plasmas against an 8 virus panel representing different genetic subtypes and phenotypes. TriMab, a mixture of three mAbs, was used as a positive control allowing comparison of the results with Phase I in a total of nine different assays. The assays used either uncloned virus produced in peripheral blood mononuclear cells (PBMCs (Virus Infectivity Assays, VIA, or Env (gp160-pseudotyped viruses (pseudoviruses, PSV produced in HEK293T cells from molecular clones or from uncloned virus. Target cells included PBMC and genetically engineered cell lines in either single- or multiple-cycle infection format. Infection was quantified by using a range of assay read-outs including extra- or intra-cellular p24 antigen detection, luciferase, beta-galactosidase or green fluorescent protein (GFP reporter gene expression. FINDINGS: Using TriMab, results of Phase I and Phase II were generally in agreement for six of the eight viruses tested and confirmed that the PSV assay is more sensitive than PBMC (p = 0.014. Comparisons with the polyclonal reagents showed that sensitivities were dependent on both virus and plasma. CONCLUSIONS: Here we further demonstrate clear differences in assay sensitivities that were dependent on both the neutralizing reagent and the virus

  19. International Network for Comparison of HIV Neutralization Assays: The NeutNet Report II

    Science.gov (United States)

    Heyndrickx, Leo; Heath, Alan; Sheik-Khalil, Enas; Alcami, Jose; Bongertz, Vera; Jansson, Marianne; Malnati, Mauro; Montefiori, David; Moog, Christiane; Morris, Lynn; Osmanov, Saladin; Polonis, Victoria; Ramaswamy, Meghna; Sattentau, Quentin; Tolazzi, Monica; Schuitemaker, Hanneke; Willems, Betty; Wrin, Terri; Fenyö, Eva Maria; Scarlatti, Gabriella

    2012-01-01

    Background Neutralizing antibodies provide markers for vaccine-induced protective immunity in many viral infections. By analogy, HIV-1 neutralizing antibodies induced by immunization may well predict vaccine effectiveness. Assessment of neutralizing antibodies is therefore of primary importance, but is hampered by the fact that we do not know which assay(s) can provide measures of protective immunity. An international collaboration (NeutNet) involving 18 different laboratories previously compared different assays using monoclonal antibodies (mAbs) and soluble CD4 (Phase I study). Methods In the present study (Phase II), polyclonal reagents were evaluated by 13 laboratories. Each laboratory evaluated nine plasmas against an 8 virus panel representing different genetic subtypes and phenotypes. TriMab, a mixture of three mAbs, was used as a positive control allowing comparison of the results with Phase I in a total of nine different assays. The assays used either uncloned virus produced in peripheral blood mononuclear cells (PBMCs) (Virus Infectivity Assays, VIA), or Env (gp160)-pseudotyped viruses (pseudoviruses, PSV) produced in HEK293T cells from molecular clones or from uncloned virus. Target cells included PBMC and genetically engineered cell lines in either single- or multiple-cycle infection format. Infection was quantified by using a range of assay read-outs including extra- or intra-cellular p24 antigen detection, luciferase, beta-galactosidase or green fluorescent protein (GFP) reporter gene expression. Findings Using TriMab, results of Phase I and Phase II were generally in agreement for six of the eight viruses tested and confirmed that the PSV assay is more sensitive than PBMC (p = 0.014). Comparisons with the polyclonal reagents showed that sensitivities were dependent on both virus and plasma. Conclusions Here we further demonstrate clear differences in assay sensitivities that were dependent on both the neutralizing reagent and the virus

  20. Detection of infliximab levels and anti-infliximab antibodies : a comparison of three different assays

    NARCIS (Netherlands)

    Casteele, N. Vande; Buurman, D. J.; Sturkenboom, M. G. G.; Kleibeuker, J. H.; Vermeire, S.; Rispens, T.; van der Kleij, D.; Gils, A.; Dijkstra, G.

    2012-01-01

    Background Formation of antibodies to infliximab (ATI) inversely correlates with functional drug levels and clinical outcome. Comparison of drug levels and anti-drug antibody monitoring is hampered by lack of standardisation. Aim To determine the correlation between three different assays for measur

  1. PCR-RFLP assays to distinguish the Western and Eastern phylogroups in wild and cultured tench Tinca tinca.

    Science.gov (United States)

    Lajbner, Z; Kotlík, P

    2011-03-01

    The tench Tinca tinca is a valued table fish native to Europe and Asia, but which is now widely distributed in many temperate freshwater regions of the world as the result of human-mediated translocations. Fish are currently being transplanted between watersheds without concern for genetic similarity to wild populations or local adaptation, and efficient phylogeographic markers are therefore urgently needed to rapidly distinguish genetically distinct geographical populations and to assess their contribution to the hatchery breeds and to the stocked wild populations. Here, we present a new method to distinguish recently discovered and morphologically undistinguishable Western and Eastern phylogroups of the tench. The method relies on PCR-RFLP assays of two independent nuclear-encoded exon-primed intron-crossing (EPIC) markers and of one mitochondrial DNA (mDNA) marker and allows the rapid identification of the Western and Eastern phylogroup and also of three geographical mtDNA clades within the Eastern phylogroup. Our method will enable researchers and fishery practitioners to rapidly distinguish genetically divergent geographical populations of the tench and will be useful for monitoring the introduction and human-mediated spread of the phylogroups in wild populations, for characterization of cultured strains and in breeding experiments.

  2. RSA/Legacy Wind Sensor Comparison. Part 1; Western Range

    Science.gov (United States)

    Short, David A.; Wheeler, Mark M.

    2006-01-01

    This report describes a comparison of data from ultrasonic and cup-and-vane anemometers on 5 wind towers at Vandenberg AFB. The ultrasonic sensors are scheduled to replace the Legacy cup-and-vane sensors under the Range Standardization and Automation (RSA) program. Because previous studies have noted differences between peak wind speeds reported by mechanical and ultrasonic wind sensors, the latter having no moving parts, the 30th and 45th Weather Squadrons wanted to understand possible differences between the two sensor types. The period-of-record was 13-30 May 2005. A total of 153,961 readings of I-minute average and peak wind speed/direction from each sensor type were used. Statistics of differences in speed and direction were used to identify 18 out of 34 RSA sensors having the most consistent performance, with respect to the Legacy sensors. Data from these 18 were used to form a composite comparison. A small positive bias in the composite RSA average wind speed increased from +0.5 kts at 15 kts, to +1 kt at 25 kts. A slightly larger positive bias in the RSA peak wind speed increased from +1 kt at 15 kts, to +2 kts at 30 kts.

  3. Comparison of In Vitro Assays in Selecting Radiotracers for In Vivo P-Glycoprotein PET Imaging

    Directory of Open Access Journals (Sweden)

    Renske M. Raaphorst

    2017-09-01

    Full Text Available Positron emission tomography (PET imaging of P-glycoprotein (P-gp in the blood-brain barrier can be important in neurological diseases where P-gp is affected, such as Alzheimer´s disease. Radiotracers used in the imaging studies are present at very small, nanomolar, concentration, whereas in vitro assays where these tracers are characterized, are usually performed at micromolar concentration, causing often discrepant in vivo and in vitro data. We had in vivo rodent PET data of [11C]verapamil, (R-N-[18F]fluoroethylverapamil, (R-O-[18F]fluoroethyl-norverapamil, [18F]MC225 and [18F]MC224 and we included also two new molecules [18F]MC198 and [18F]KE64 in this study. To improve the predictive value of in vitro assays, we labeled all the tracers with tritium and performed bidirectional substrate transport assay in MDCKII-MDR1 cells at three different concentrations (0.01, 1 and 50 µM and also inhibition assay with P-gp inhibitors. As a comparison, we used non-radioactive molecules in transport assay in Caco-2 cells at a concentration of 10 µM and in calcein-AM inhibition assay in MDCKII-MDR1 cells. All the P-gp substrates were transported dose-dependently. At the highest concentration (50 µM, P-gp was saturated in a similar way as after treatment with P-gp inhibitors. Best in vivo correlation was obtained with the bidirectional transport assay at a concentration of 0.01 µM. One micromolar concentration in a transport assay or calcein-AM assay alone is not sufficient for correct in vivo prediction of substrate P-gp PET ligands.

  4. A comparison of sugar indicators enables a universal high-throughput sugar-1-phosphate nucleotidyltransferase assay.

    Science.gov (United States)

    Moretti, Rocco; Thorson, Jon S

    2008-06-15

    A systematic comparison of six sugar indicators for their sensitivity, specificity, cross-reactivity, and suitability in the context of crude lysates revealed para-hydroxybenzoic acid hydrazide (pHBH) to be best suited for application in a plate-based phosphatase-assisted universal sugar-1-phosphate nucleotidyltransferase assay. The addition of a general phosphatase to nucleotidyltransferase reaction aliquots enabled the conversion of remaining sugar-1-phosphate to free sugar, the concentration of which could be rapidly assessed via the pHBH assay. The assay was validated using the model glucose-1-phosphate thymidylyltransferase from Salmonella enterica (RmlA) and compared favorably with a previously reported HPLC assay. This coupled discontinuous assay is quantitative, high throughput, and robust; relies only on commercially available enzymes and reagents; does not require chromatography, specialized detectors (e.g., mass or evaporative light scattering detectors), or radioisotopes; and is capable of detecting less than 5 nmol of sugar-1-phosphate. It is anticipated that this high-throughput assay system will greatly facilitate nucleotidyltransferase mechanistic and directed evolution/engineering studies.

  5. The Multispot rapid HIV-1/HIV-2 differentiation assay is comparable with the Western blot and an immunofluorescence assay at confirming HIV infection in a prospective study in three regions of the United States.

    Science.gov (United States)

    Pandori, Mark W; Westheimer, Emily; Gay, Cindy; Moss, Nicholas; Fu, Jie; Hightow-Weidman, Lisa B; Craw, Jason; Hall, Laura; Giancotti, Francesca R; Mak, Mae Ling; Madayag, Carmela; Tsoi, Benjamin; Louie, Brian; Patel, Pragna; Owen, S Michele; Peters, Philip J

    2013-12-01

    A new HIV diagnostic algorithm has been proposed which replaces the use of the HIV-1 Western blot and HIV-1 immunofluorescence assays (IFA) as the supplemental test with an HIV-1/HIV-2 antibody differentiation assay. To compare an FDA-approved HIV-1/HIV-2 antibody differentiation test (Multispot) as a confirmatory test with the HIV-1 Western blot and IFA. Participants were screened with an HIV-1/HIV-2 combination Antigen/Antibody (Ag/Ab) screening assay. Specimens with repeatedly reactive results were tested with Multispot and either Western blot or IFA. Specimens with discordant screening and confirmatory results were resolved with HIV-1 RNA testing. Individuals (37,876) were screened for HIV infection and 654 (1.7%) had a repeatedly reactive Ag/Ab assay result. On Multispot, 554 (84.7%) were HIV-1 reactive, 0 (0%) were HIV-2 reactive, 1 (0.2%) was reactive for both HIV-1 and HIV-2 (undifferentiated), 9 (1.4%) were HIV-1 indeterminate, and 90 (13.8%) were non-reactive. HIV-1 RNA was detected in 47/90 Multispot non-reactive (52.2%) specimens. Among specimens confirmed to have HIV infection (true positives), Multispot and Western blot detected HIV-1 antibody in a similar proportion of cases (93.7% vs. 94.4% respectively) while Multispot and IFA also detected HIV-1 antibody in a similar proportion of cases (84.5% vs. 83.4% respectively). In this study, Multispot confirmed HIV infections at a similar proportion to Western blot and IFA. Multispot, Western blot, and IFA, however, did not confirm all of the reactive Ag/Ab assay results and underscores the importance of HIV NAT testing to resolve discordant screening and confirmatory results. Copyright © 2013 The Authors. Published by Elsevier B.V. All rights reserved.

  6. Comparison of antibody responses to human papillomavirus vaccination as measured by three assays

    Directory of Open Access Journals (Sweden)

    Hilary Ann Robbins

    2014-01-01

    Full Text Available Background: Different assays, including the competitive Luminex immunoassay (cLIA, secreted alkaline phosphatase neutralization assay (SEAP-NA, and virus-like particle-based ELISA, are commonly used to measure antibody responses after human papillomavirus (HPV vaccination. Direct assay comparisons aid interpretation of immunogenicity data evaluated by different assays. Methods: We compared cLIA to SEAP-NA and ELISA among 51 HPV16/18-vaccinated women enrolled in the Costa Rica Vaccine Trial. We tested replicate serum samples collected at months 0, 1, and 12 by HPV16/18 cLIA, SEAP-NA, and ELISA. For a subset (N=10, we further tested month 24 and 36 samples. We calculated seroprevalence estimates and Spearman rank correlation coefficients comparing cLIA to SEAP-NA and ELISA.Results: After one vaccine dose, seroprevalence by SEAP-NA and ELISA was 100% (both HPV16 and HPV18, and by cLIA was 96% (95% CI 87%-100% for HPV16 and 71% (95% CI 56%-83% for HPV18. Seroprevalence was 100% by all assays after 3 doses. Correlation between assays was high after one vaccine dose (cLIA/SEAP-NA ρ=0.91 (HPV16 and ρ=0.86 (HPV18; cLIA/ELISA ρ=0.84 (HPV16 and ρ=0.74 (HPV18; all p<0.001 and remained high through month 36. Ratios of mean antibody levels to seropositivity cutoffs at month 36 were lower for cLIA than for SEAP-NA or ELISA, particularly for HPV18 (HPV18 ratio for cLIA 1.9, SEAP-NA 3.5, ELISA 3.4.Conclusion: Though correlation between cLIA and SEAP-NA/ELISA is high and stable after vaccination, the assays differ in scale and sensitivity, with notable differences after 1 vaccine dose and for HPV18. Our results demonstrate that comparisons of antibody responses to HPV vaccination measured by different assays are approximate, and must consider biological and technical differences between assays.

  7. A Comparison of In-House Real-Time LAMP Assays with a Commercial Assay for the Detection of Pathogenic Bacteria

    OpenAIRE

    Deguo Wang; Yongzhen Wang; Fugang Xiao; Weiyun Guo; Yongqing Zhang; Aiping Wang; Yanhong Liu

    2015-01-01

    Molecular detection of bacterial pathogens based on LAMP methods is a faster and simpler approach than conventional culture methods. Although different LAMP-based methods for pathogenic bacterial detection are available, a systematic comparison of these different LAMP assays has not been performed. In this paper, we compared 12 in-house real-time LAMP assays with a commercialized kit (Isothermal Master Mix) for the detection of Listeria monocytogenes, Salmonella spp, Staphylococcus aureus, E...

  8. Technical note: comparison of the PrestoBlue and LDH release assays with the MTT assay for skin viability assessment.

    Science.gov (United States)

    Gaucher, Sonia; Jarraya, Mohamed

    2015-09-01

    MTT assay is the gold standard for assessing skin sample viability but it is time-consuming. Here we compared the MTT test with two other assays for the assessment of skin viability. The MTT, PrestoBlue (colorimetric method) and LDH release assays were applied to fresh and cryopreserved skin. Skin viability was considered proportional to the optical density values of the relevant analytes. PrestoBlue did not reliably distinguish between fresh and cryopreserved skin. The LDH release assay did not allow us to establish a viability index. We recommend the MTT assay for assessing skin viability.

  9. Viability-qPCR for detecting Legionella: Comparison of two assays based on different amplicon lengths.

    Science.gov (United States)

    Ditommaso, Savina; Giacomuzzi, Monica; Ricciardi, Elisa; Zotti, Carla M

    2015-08-01

    Two different real-time quantitative PCR (PMA-qPCR) assays were applied for quantification of Legionella spp. by targeting a long amplicon (approx 400 bp) of 16S rRNA gene and a short amplicon (approx. 100 bp) of 5S rRNA gene. Purified DNA extracts from pure cultures of Legionella spp. and from environmental water samples were quantified. Application of the two assays to quantify Legionella in artificially contaminated water achieved that both assays were able to detect Legionella over a linear range of 10 to 10(5) cells ml(-1). A statistical analysis of the standard curves showed that both assays were linear with a good correlation coefficient (R(2) = 0.99) between the Ct and the copy number. Amplification with the reference assay was the most effective for detecting low copy numbers (1 bacterium per PCR mixture). Using selective quantification of viable Legionella by the PMA-qPCR method we obtained a greater inhibition of the amplification of the 400-bp 16S gene fragment (Δlog(10) = 3.74 ± 0.39 log(10) GU ml(-1)). A complete inhibition of the PCR signal was obtained when heat-killed cells in a concentration below 1 × 10(5) cells ml(-1) were pretreated with PMA. Analysing short amplicon sizes led to only 2.08 log reductions in the Legionella dead-cell signal. When we tested environmental water samples, the two qPCR assays were in good agreement according to the kappa index (0.741). Applying qPCR combined with PMA treatment, we also obtained a good agreement (kappa index 0.615). The comparison of quantitative results shows that both assays yielded the same quantification sensitivity (mean log = 4.59 vs mean log = 4.31).

  10. Comparison of two rapid assays for Clostridium difficile Common antigen and a C difficile toxin A/B assay with the cell culture neutralization assay.

    Science.gov (United States)

    Reller, Megan E; Alcabasa, Romina C; Lema, Clara A; Carroll, Karen C

    2010-01-01

    We compared 3 rapid assays for Clostridium difficile with a cell culture cytotoxicity neutralization assay (CCNA). Of 600 stool samples, 46 were positive for toxigenic C difficile. Both rapid common antigen assays were highly sensitive (91.3%-100%) and, therefore, were appropriate screening tests. The rapid toxin assay had poor sensitivity (61%) but excellent specificity (99.3%). Testing stools for glutamate dehydrogenase (step 1) and those positive with a rapid toxin assay (step 2) would correctly classify 81% of submitted specimens within 2 hours, including during periods of limited staffing (evenings, nights, and weekends). CCNA could then be used as a third step to test rapid toxin-negative samples, thereby providing a final result for the remaining 19% of samples by 48 to 72 hours. The use of rapid assays as outlined could enhance timely diagnosis of C difficile.

  11. Comparison between fibroblast wound healing and cell random migration assays in vitro.

    Science.gov (United States)

    Ascione, Flora; Vasaturo, Angela; Caserta, Sergio; D'Esposito, Vittoria; Formisano, Pietro; Guido, Stefano

    2016-09-10

    Cell migration plays a key role in many biological processes, including cancer growth and invasion, embryogenesis, angiogenesis, inflammatory response, and tissue repair. In this work, we compare two well-established experimental approaches for the investigation of cell motility in vitro: the cell random migration (CRM) and the wound healing (WH) assay. In the former, extensive tracking of individual live cells trajectories by time-lapse microscopy and elaborate data processing are used to calculate two intrinsic motility parameters of the cell population under investigation, i.e. the diffusion coefficient and the persistence time. In the WH assay, a scratch is made in a confluent cell monolayer and the closure time of the exposed area is taken as an easy-to-measure, empirical estimate of cell migration. To compare WH and CRM we applied the two assays to investigate the motility of skin fibroblasts isolated from wild type and transgenic mice (TgPED) overexpressing the protein PED/PEA-15, which is highly expressed in patients with type 2 diabetes. Our main result is that the cell motility parameters derived from CRM can be also estimated from a time-resolved analysis of the WH assay, thus showing that the latter is also amenable to a quantitative analysis for the characterization of cell migration. To our knowledge this is the first quantitative comparison of these two widely used techniques. Copyright © 2016 Elsevier Inc. All rights reserved.

  12. Comparison of total protein concentration in skeletal muscle as measured by the Bradford and Lowry assays.

    Science.gov (United States)

    Seevaratnam, Rajini; Patel, Barkha P; Hamadeh, Mazen J

    2009-06-01

    The Lowry and Bradford assays are the most commonly used methods of total protein quantification, yet vary in several aspects. To date, no comparisons have been made in skeletal muscle. We compared total protein concentrations of mouse red and white gastrocnemius, reagent stability, protein stability and range of linearity using both assays. The Lowry averaged protein concentrations 15% higher than the Bradford with a moderate correlation (r = 0.36, P = 0.01). However, Bland-Altman analysis revealed considerable bias (15.8 +/- 29.7%). Both Lowry reagents and its protein-reagent interactions were less stable over time than the Bradford. The linear range of concentration was smaller for the Lowry (0.05-0.50 mg/ml) than the Bradford (0-2.0 mg/ml). We conclude that the Bradford and Lowry measures of total protein concentration in skeletal muscle are not interchangeable. The Bradford and Lowry assays have various strengths and weaknesses in terms of substance interference and protein size. However, the Bradford provides greater reagent stability, protein-reagent stability and range of linearity, and requires less time to analyse compared to the Lowry assay.

  13. Discrepancies between a new highly sensitive Toxoplasma gondii ELISA assay and other reagents: interest of Toxo IgG Western blot.

    Science.gov (United States)

    Leslé, F; Touafek, F; Fekkar, A; Mazier, D; Paris, L

    2011-10-01

    Immunodiagnostic assays are commonly used to screen for maternal toxoplasmic seroconversion during pregnancy. The introduction to the market of a new highly sensitive IgG assay, the Elecsys Toxo IgG test, has resulted in discrepancy issues with other immunoassays because of a lack of standardisation. Western blot appears to be a good alternative gold standard to the dye test, as the latter is not routinely available. For the present prospective study, we compared the analytical performances of two immunoassays, Elecsys Toxo IgG (Roche Diagnostics) and Platelia Toxo IgG (Bio-Rad, Marnes la Coquette, France), to Toxo II IgG Western blot (LDBio, Lyon, France) using 231 consecutive sera with low or equivocal IgG titres. Of these 231 sera, 213 presented discrepancies, which showed the importance of a confirmation test. Of the Elecsys Toxo IgG-positive results, 100% were confirmed by the Western blot with a positive threshold of 30 IU/ml for Elecsys; in the equivocal area (1-30 IU/ml), Western blot is negative in 54% of cases. Our results suggest that the lower diagnostic cut-off of Platelia Toxo IgG should be further reduced. Our study indirectly confirms that monitoring, especially for pregnant women, must be done in the same laboratory using the same technique. The ability to diagnose very early seroconversion using Western blot merits further study.

  14. Dose estimation using dicentric chromosome assay and cytokinesis block micronucleus assay: comparison between manual and automated scoring in triage mode.

    Science.gov (United States)

    De Amicis, Andrea; De Sanctis, Stefania; Di Cristofaro, Sara; Franchini, Valeria; Regalbuto, Elisa; Mammana, Giacomo; Lista, Florigio

    2014-06-01

    In cases of an accidental overexposure to ionizing radiation, it is essential to estimate the individual absorbed dose of a potentially radiation-exposed person. For this purpose, biological dosimetry can be performed to confirm, complement or even replace physical dosimetry when this proves to be unavailable. The most validated biodosimetry techniques for dose estimation are the dicentric chromosome assay, the "gold standard" for individual dose assessment, and cytokinesis-block micronucleus assay. However, both assays are time consuming and require skilled scorers. In case of large-scale accidents, different strategies have been developed to increase the throughput of cytogenetic service laboratories. These are the decrease of cell numbers to be scored for triage dosimetry; the automation of procedures including the scoring of, for example, aberrant chromosomes and micronuclei; and the establishment of laboratory networks in order to enable mutual assistance if necessary. In this study, the authors compared the accuracy of triage mode biodosimetry by dicentric chromosome analysis and the cytokinesis block micronucleus assay performing both the manual and the automated scoring mode. For dose estimation using dicentric chromosome assay of 10 blind samples irradiated up to 6.4 Gy of x-rays, a number of metaphase spreads were analyzed ranging from 20 up to 50 cells for the manual and from 20 up to 500 cells for the automatic scoring mode. For dose estimation based on the cytokinesis block micronucleus assay, the micronucleus frequency in both 100 and 200 binucleated cells was determined by manual and automatic scoring. The results of both assays and scoring modes were compared and analyzed considering the sensitivity, specificity, and accuracy of dose estimation with regard to the discrimination power of clinically relevant binary categories of exposure doses.

  15. A comparative study of Toxoplasma gondii seroprevalence in mink using a modified agglutination test, a Western blot, and enzyme-linked immunosorbent assays.

    Science.gov (United States)

    Gu, Yi; Wang, Zedong; Cai, Yufeng; Li, Xiaoxing; Wei, Feng; Shang, Limin; Li, Jiping; Liu, Quan

    2015-09-01

    Toxoplasma gondii can infect almost all warm-blooded animals, and many serological methods have been developed to detect T. gondii infection in a variety of animal species. In the present study, the seroprevalence of T. gondii infection in farmed mink in northeast China was determined using the modified agglutination test (MAT), a Western blot (WB), and 3 enzyme-linked immunosorbent assays (ELISAs) with protein A/G conjugate, using either of 2 recombinant dense granule antigens, GRA1 and GRA7, or Toxoplasma soluble antigens (TSA). There was no significant difference between the detection results of the GRA1-, GRA7-, and TSA-ELISAs and WB (McNemar chi-square, P > 0.05), but a significant difference was observed between MAT and WB (P < 0.05). A near perfect agreement (97.0%) was found between the GRA7-ELISA and WB (κ = 0.83), and a substantial agreement (92.4-93.1%) was observed in the TSA- and GRA1-ELISAs (κ = 0.68-0.73). The GRA7-ELISA showed the highest sensitivity and specificity, and the lowest false-positive and negative rates, while the MAT gave both a low sensitivity and frequent false positives in comparison to the WB. Receiver operating characteristic analysis revealed the largest area under curve of 0.85 (95% confidence interval: 0.74-0.96), and the highest relative sensitivity (72.7%) and specificity (99.0%) for a cutoff value of 0.19 in the GRA7-ELISA. These results indicate that the GRA7-ELISA is suitable for detection of T. gondii infection in mink and that MAT should be used with caution.

  16. A comparison on urbanization of two mega-polises in western China in recent 20 years

    Institute of Scientific and Technical Information of China (English)

    MA Ze-zhong

    2007-01-01

    Based on a comparison of the spatial extension modulus and speed of the two mega-polis cities in western China, Chongqing and Chengdu, the differences of urbanization characteristics of the two cities were analyzed. The results show that the two cities have different spatial extension modulus and extension speed. The extension speed of Chongqing, the largest mountainous city of China, was slow and its extension direction was along the Yangtze River valley and the Jialing River valley. Chengdu, the largest plain city of western China, had a faster extension speed than Chongqing and a concentric circles extent modulus. The spatial extent modulus and speed were controlled by the natural condition, especially the topography, the policies, the economic development level and the investment of the state central government.

  17. The epidemiology of tick-borne haemoparasites as determined by the reverse line blot hybridization assay in an intensively studied cohort of calves in western Kenya.

    Science.gov (United States)

    Njiiri, Nyawira E; Bronsvoort, B Mark deC; Collins, Nicola E; Steyn, Helena C; Troskie, Milana; Vorster, Ilse; Thumbi, S M; Sibeko, Kgomotso P; Jennings, Amy; van Wyk, Ilana Conradie; Mbole-Kariuki, Mary; Kiara, Henry; Poole, E Jane; Hanotte, Olivier; Coetzer, Koos; Oosthuizen, Marinda C; Woolhouse, Mark; Toye, Philip

    2015-05-30

    The development of sensitive surveillance technologies using PCR-based detection of microbial DNA, such as the reverse line blot assay, can facilitate the gathering of epidemiological information on tick-borne diseases, which continue to hamper the productivity of livestock in many parts of Africa and elsewhere. We have employed a reverse line blot assay to detect the prevalence of tick-borne parasites in an intensively studied cohort of indigenous calves in western Kenya. The calves were recruited close to birth and monitored for the presence of infectious disease for up to 51 weeks. The final visit samples from 453 calves which survived for the study period were analyzed by RLB. The results indicated high prevalences of Theileria mutans (71.6%), T. velifera (62.8%), Anaplasma sp. Omatjenne (42.7%), A. bovis (39.9%), Theileria sp. (sable) (32.7%), T. parva (12.9%) and T. taurotragi (8.5%), with minor occurrences of eight other haemoparasites. The unexpectedly low prevalence of the pathogenic species Ehrlichia ruminantium was confirmed by a species-specific PCR targeting the pCS20 gene region. Coinfection analyses of the seven most prevalent haemoparasites indicated that they were present as coinfections in over 90% of the cases. The analyses revealed significant associations between several of the Theileria parasites, in particular T. velifera with Theileria sp. sable and T. mutans, and T. parva with T. taurotragi. There was very little coinfection of the two most common Anaplasma species, although they were commonly detected as coinfections with the Theileria parasites. The comparison of reverse line blot and serological results for four haemoparasites (T. parva, T. mutans, A. marginale and B. bigemina) indicated that, except for the mostly benign T. mutans, indigenous cattle seem capable of clearing infections of the three other, pathogenic parasites to below detectable levels. Although the study site was located across four agroecological zones, there was

  18. Association of clinical and genetical features in FMF with focus on MEFV strip assay sensitivity in 452 children from western Anatolia, Turkey.

    Science.gov (United States)

    Ozturk, Can; Halicioglu, Oya; Coker, Işil; Gulez, Nesrin; Sutçuoglu, Sumer; Karaca, Neslihan; Aksu, Guzide; Kutukculer, Necil

    2012-03-01

    The aim of this study was to determine the relationship between clinical findings and the most common mutated alleles of MEFV gene in a childhood population and to determine the sensitivity of the 12-mutation-strip assay test in familial Mediterranean fever (FMF). Records of 452 FMF children living in western Anatolia, Turkey, (12.3 ± 4.7 years mean) were retrospectively reviewed. Of the 408 patients who met the Tel-Hashomer criteria, 364 were classified into two main groups (two-mutant/one-mutant allele) either of which had three subgroups. The two-mutant allele frequency was 51% and one-mutant allele 38%; 1% had complex-mutant alleles and 10% no mutant-alleles. The mean severity score was 8.3 ± 2.5. Most common clinical features were fever (81.9%), abdominal pain (86.3%) and myalgia (58.8%), and the least common ones: diarrhea (1.7%), protracted febrile myalgia (1.2%) and acute orchitis (1.5%). We detected 33 different genotypes of the MEFV gene: the most common mutant allele was M694V followed by symptomatic allele mutation of E148Q. Although not significantly associated with clinical findings, P369S mutation was not rare (7.5%). Phenotype-genotype correlation revealed that patients with two-allele mutations had more severe clinical presentation and high constipation rate (22.5%); 32.6% of patients with M694V/M694V had splenomegaly. Acute orchitis and protracted febrile myalgia as rare clinical findings were more common in M694V homozygotes. Comparisons of clinical findings among patients with one-mutation allele were made for the first time, but no significant association was found. Positive predictive value of strip assay screening for 12 mutations was recorded as 89%. We suggest that whole sequence analysis for supportive diagnosis of FMF should be performed for selected patients only.

  19. Development of a TaqMan assay for the six major genotypes of hepatitis C virus: Comparison with commercial assays

    DEFF Research Database (Denmark)

    Engle, Ronald E; Russell, Rodney S; Purcell, Robert H

    2008-01-01

    A quantitative real-time PCR assay was developed that detects genomic RNA from reference strains representing the six major genotypes of hepatitis C virus (HCV) with equal sensitivity and accurately measured HCV RNA in JFH1 HCV-infected Huh7.5 cells. The method is indirectly calibrated to the first...

  20. Comparison of three human papillomavirus DNA assays and one mRNA assay in women with abnormal cytology

    DEFF Research Database (Denmark)

    Rebolj, Matejka; Lynge, Elsebeth; Ejegod, Ditte;

    2014-01-01

    OBJECTIVE: To compare the clinical characteristics of four human papillomavirus (HPV) assays: hybrid capture 2 (HC2), cobas, CLART, and APTIMA in Danish women with abnormal cytology. METHODS: SurePath samples from 367 consecutive women from Copenhagen, with atypical squamous cells of undetermined...

  1. A systematic comparison of three commercial estrogen receptor assays in a single clinical outcome breast cancer cohort.

    Science.gov (United States)

    Kornaga, Elizabeth N; Klimowicz, Alexander C; Guggisberg, Natalia; Ogilvie, Travis; Morris, Don G; Webster, Marc; Magliocco, Anthony M

    2016-08-01

    Breast cancers are routinely assessed for estrogen receptor status using immunohistochemical assays to assist in patient prognosis and clinical management. Specific assays vary between laboratories, and several antibodies have been validated and recommended for clinical use. As numerous factors can influence assay performance, many laboratories have opted for ready-to-use assays using automated stainers to improve reproducibility and consistency. Three commonly used autostainer vendors-Dako, Leica, and Ventana-all offer such estrogen receptor assays; however, they have never been directly compared. Here, we present a systematic comparison of three platform-specific estrogen receptor ready-to-use assays using a retrospective, tamoxifen-treated, breast cancer cohort from patients who were treated in Calgary, Alberta, Canada from 1985 to 2000. We found all assays showed good intra-observer agreement. Inter-observer pathological scoring showed some variability: Ventana had the strongest agreement followed closely by Dako, whereas Leica only showed substantial agreement. We also analyzed each estrogen receptor assay with respect to 5-year disease-free survival, and found that all performed similarly in univariate and multivariate models. Determination of measures of test performance found that the Leica assay had a lower negative predictive value than Dako or Ventana, compared with the original ligand-binding assay, while other measures-sensitivity, specificity, positive predictive value, and accuracy-were comparable between the three ready-to-use assays. When comparing against disease-free survival, the difference in negative predictive value between the vendor assays were not as extreme, but Dako and Ventana still performed slightly better than Leica. Despite some discordance, we found that all ready-to-use assays were comparable with or superior to the ligand-binding assay, endorsing their continued use. Our analysis also allowed for exploration of estrogen receptor

  2. Comparison of antibacterial activities of root-end filling materials by an agar diffusion assay and Alamar blue assay

    Directory of Open Access Journals (Sweden)

    Tsui-Hsien Huang

    2012-12-01

    Conclusion: We concluded that both the agar diffusion test and Alamar blue assay gave comparable findings of assessing the antimicrobial activity present in root-end filling materials. No antimicrobial activity was detected for mineral trioxide aggregate, calcium silicate cement, or amalgam after coming into contact with S. mutans, S. sanguinis and E. coli. IRM showed high antimicrobial activity against both S. sanguinis and E. coli.

  3. Comparison of Acute Toxicity of Algal Metabolites Using Bioluminescence Inhibition Assay

    Directory of Open Access Journals (Sweden)

    Hansa Jeswani

    2015-01-01

    Full Text Available Microalgae are reported to degrade hazardous compounds. However, algae, especially cyanobacteria are known to produce secondary metabolites which may be toxic to flora, fauna and human beings. The aim of this study was selection of an appropriate algal culture for biological treatment of biomass gasification wastewater based on acute toxicity considerations. The three algae that were selected were Spirulina sp., Scenedesmus abundans and a fresh water algal consortium. Acute toxicity of the metabolites produced by these algal cultures was tested at the end of log phase using the standard bioluminescence inhibition assay based on Vibrio fischeri NRRLB 11174. Scenedesmus abundans and a fresh water algal consortium dominated by cyanobacteria such as Phormidium, Chroococcus and Oscillatoria did not release much toxic metabolites at the end of log phase and caused only about 20% inhibition in bioluminescence. In comparison, Spirulina sp. released toxic metabolites and caused 50% bioluminescence inhibition at 3/5 times dilution of the culture supernatant (EC50.

  4. Comparison of histopathology and PCR based assay for detection of experimentally induced toxoplasmosis in murine model

    Institute of Scientific and Technical Information of China (English)

    Vikrant Sudan; A K Tewari; R Singh; Harkirat Singh

    2015-01-01

    Objective:To compare histopathology and PCR based detection in diagnosis of experimentally induced toxoplasmosis of RH human strain of the parasite in murine models. Methods:A comparison of histopathology and PCR based detection was done to diagnose experimentally induced toxoplasmosis in ten inbred swiss albino mice after intraperitoneal inoculation of 100 tachyzoites of laboratory mantained human RH strain of the parasite. Tissue samples from lung, liver, spleen, brain, heart and kidney were taken and processed for histopathological examination while all the samples also were subjected to PCR, using primers directed to the multicopy of SAG 3 gene, in dublicates. Results: Histopathology revealed presence of tachyzoites only in liver while along with lung, liver, spleen and brain tissue yielded desired positive PCR amplicons. Conclusions:The SAG 3 based PCR is able to diagnose toxoplasmosis in those tissues which are declared negative by histopathological assay.

  5. Interlaboratory comparison of the dicentric chromosome assay for radiation biodosimetry in mass casualty events.

    Science.gov (United States)

    Wilkins, Ruth C; Romm, Horst; Kao, Tzu-Cheg; Awa, Akio A; Yoshida, Mitsuaki A; Livingston, Gordon K; Jenkins, Mark S; Oestreicher, Ursula; Pellmar, Terry C; Prasanna, Pataje G S

    2008-05-01

    This interlaboratory comparison validates the dicentric chromosome assay for assessing radiation dose in mass casualty accidents and identifies the advantages and limitations of an international biodosimetry network. The assay's validity and accuracy were determined among five laboratories following the International Organization for Standardization guidelines. Blood samples irradiated at the Armed Forces Radiobiology Research Institute were shipped to all laboratories, which constructed individual radiation calibration curves and assessed the dose to dose-blinded samples. Each laboratory constructed a dose-effect calibration curve for the yield of dicentrics for (60)Co gamma rays in the 0 to 5-Gy range, using the maximum likelihood linear-quadratic model, Y = c + alphaD + betaD(2). For all laboratories, the estimated coefficients of the fitted curves were within the 99.7% confidence intervals (CIs), but the observed dicentric yields differed. When each laboratory assessed radiation doses to four dose-blinded blood samples by comparing the observed dicentric yield with the laboratory's own calibration curve, the estimates were accurate in all laboratories at all doses. For all laboratories, actual doses were within the 99.75% CI for the assessed dose. Across the dose range, the error in the estimated doses, compared to the physical doses, ranged from 15% underestimation to 15% overestimation.

  6. Comparison of five commercial enzyme-linked immunosorbent assays for detection of antibodies to Bordetella pertussis.

    Science.gov (United States)

    Kösters, K; Riffelmann, M; Dohrn, B; von König, C H

    2000-05-01

    Measuring antibodies to Bordetella pertussis antigens is mostly done by enzyme-linked immunosorbent assays (ELISAs). We compared the performance of five commercially available ELISA kits with the help of 65 serum specimens which were repetitively tested for evaluation of the kits. The specimens contained 20 paired serum samples from patients with clinical pertussis, 15 samples were from children vaccinated with a diphtheria-tetanus-acellular pertussis vaccine, seven specimens were taken from an interlaboratory comparison of ELISAs, and there were three reference preparations from the Food and Drug Administration's (FDA's) Laboratory of Pertussis and from our laboratory. Reference values were obtained from the FDA or from results obtained with an in-house ELISA. Commercial ELISAs were compared with respect to their reproducibility and variability, their ability to detect significant titer rises in paired serum samples, their ability to detect an immune response after vaccination, and the comparability of semiquantitative and quantitative results. Reproducibility was generally good (>89%), intra-assay variation ranged from 2.4 to 28.7%, and indeterminate results were recorded in up to 18.5% of all specimens. Most kits correctly identified the antibody response to an acellular pertussis vaccine. None of the commercial kits identified all cases of pertussis correctly, and the sensitivity ranged between 60 and 95%. All five commercial ELISAs showed great discrepancies when comparing semiquantitative results and contained obviously different antigen preparations. Our data suggest that the five commercial ELISAs tested here need further improvement and standardization.

  7. A comparison between medicine from an African (Ubuntu and Western philosophy

    Directory of Open Access Journals (Sweden)

    ED Prinsloo

    2001-09-01

    Full Text Available I consider the Ubuntu way of caring for the sick in terms of the Ubuntu world-view by systematizing the scattered views. I argue that this world-view is underpinned by the regulative concept of sharing and that caring in Ubuntuthinking can only be understood correctly in terms of sharing. I substantiate my exposition in terms of what Africans themselves claim Ubuntu is and relate its meaning to African thinking in general. I consider the uniqueness of this world-view by showing how an African thinker compares it to Western world-views on causality and critically consider these comparisons. I apply this world-view to African medicine and evaluate the Ubuntu idea of causes in medicine in comparison with causality in Western thinking by considering the two frameworks of medical care in terms of their viability respectively. I conclude that causal patterns in medicine are controversial in both thinkings but argue that it sets the framework for intercultural communication that can lead both to a better understanding of each other and to some positive developments in medicine. These ways of dealing with the topic represents the significance of this article as an addition to existing knowledge.

  8. Comparison of enzyme-linked immunosorbent assay test with immunoblot assay in the diagnosis of pemphigus in Indian patients

    Directory of Open Access Journals (Sweden)

    Khandpur Sujay

    2010-01-01

    Full Text Available Background: The diagnosis of pemphigus vulgaris (PV and pemphigus foliaceous (PF rests upon clinical, histological and immunofluorescence features. Enzyme-linked immunosorbent assay (ELISA test and immunoblot (IB assay have shown variable sensitivity and specificity. Aims: We compared the utility of ELISA and IB in pemphigus patients. Methods: Sixty-six pemphigus cases (PV-54, PF-12 and 72 controls (other vesicobullous disorders and healthy controls were inducted. ELISA for anti-Dsg 3 and 1 antibodies and IB assay were performed. Results: On ELISA, both mean anti-Dsg 1 and 3 titers were raised in PV and PF. Mean anti-Dsg 1 in mucocutaneous PV was significantly higher than in mucosal PV and mean anti-Dsg 3 was significantly raised in PV than in PF. Anti-Dsg 1 and 3 in the control group were negative. Sensitivity and specificity of ELISA in PV was 98.14% and 90.5% while in PF it was 91.6% and 61.1%, respectively.On IB in PV, 36 cases (66.67% showed the 130 kDa and 160 kDa antigen bands, 12 (22.2% only the 130 kDa and six (11.1% only the 160 kDa band. Eight of the nine pure mucosal cases (88.8% showed only the 130 kDa. In PF, only the 160 kDa antigen was detected. These antigens were not identified in the control group. Sensitivity and specificity of IB in PV was 88.9% and 100% and in PF it was 100% and 95.2%, respectively. Conclusion: Both tests could differentiate pemphigus from other dermatoses, including other blistering disorders. ELISA could not make a distinction between PV and PF or between the various clinical phenotypes of PV. IB differentiated between PV and PF and the different clinical variants of PV.

  9. Comparison of genotoxicity of textile dyestuffs in Salmonella mutagenicity assay, in vitro micronucleus assay, and single cell gel/comet assay.

    Science.gov (United States)

    Wollin, Klaus-M; Gorlitz, Bernd-D

    2004-01-01

    The mutagenicity of textile dyes is an important consideration for the assurance of consumer protection and work safety. The mutagenicity testing of textile dyestuffs is crucial for accurately predicting health risks for consumers and workers exposed to dyes. Unfortunately, these data are often lacking. We studied the genotoxic activity of ten selected commercial textile dyestuffs, which are made up of mixtures of azo dyes and azo metal complex dyes as well as two anthraquinone dyestuffs. We used the Salmonella mutagenicity assay and cultured human keratinocytes (HaCaT cell line). In the S. typhimurium strain TA98, with and without S9, eight often dyestuffs investigated, and in strain TA 100, with and without S9, six often dyes caused frameshift mutations and base-pair substitutions in the dose range of 1-5000 microg/plate in a dose-related manner. All dyes, including those negative in the Salmonella mutagenicity assay, induced clastogenic effects in the in vitro micronucleus (MN) test in HaCaT cells as direct-acting mutagens in the concentration range of 5-150 microg/mL and with maximum MN frequencies between 1.1 and 7.2%, compared to negative controls that showed 0.2-0.4% MN cells. In the single cell gel/comet assay, all ten dyestuffs investigated caused DNA damage in HaCaT keratinocytes. The alkaline (pH >13) version used is capable of detecting DNA single strand breaks, alkali-labile sites, and DNA-DNA/DNA-protein cross-linking. Under the conditions of these screening tests, the textile dyes investigated are direct-acting genotoxic substances. The HaCaT cells testing protocol proposed has been shown to be an appropriate test system for evaluating mutagenicity of textile dyes on a base level.

  10. A Comparison of In-House Real-Time LAMP Assays with a Commercial Assay for the Detection of Pathogenic Bacteria.

    Science.gov (United States)

    Wang, Deguo; Wang, Yongzhen; Xiao, Fugang; Guo, Weiyun; Zhang, Yongqing; Wang, Aiping; Liu, Yanhong

    2015-05-25

    Molecular detection of bacterial pathogens based on LAMP methods is a faster and simpler approach than conventional culture methods. Although different LAMP-based methods for pathogenic bacterial detection are available, a systematic comparison of these different LAMP assays has not been performed. In this paper, we compared 12 in-house real-time LAMP assays with a commercialized kit (Isothermal Master Mix) for the detection of Listeria monocytogenes, Salmonella spp, Staphylococcus aureus, Escherichia coli O157, E. coli O26, E. coli O45, E. coli O103, E. coli O111, E. coli O121, E. coli O145 and Streptococcus agalactiae. False-positive results were observed in all 12 in-house real-time LAMP assays, while all the negative controls of Isothermal Master Mix remained negative after amplification. The detection limit of Isothermal Master Mix for Listeria monocytogenes, Salmonella spp, Staphylococcus aureus, Escherichia coli O157, E. coli O26, E. coli O45, E. coli O103, E. coli O111, E. coli O121 and Streptococcus agalactiae was 1 pg, whereas the sensitivity of the commercialized kit for E. coli O145 was 100 pg. In conclusion, the 12 in-house real-time LAMP assays were impractical to use, while the commercialized kit Isothermal Master Mix was useful for the detection of most bacterial pathogens.

  11. Lake isotope records of the 8200-year cooling event in western Ireland: Comparison with model simulations

    Science.gov (United States)

    Holmes, Jonathan A.; Tindall, Julia; Roberts, Neil; Marshall, William; Marshall, Jim D.; Bingham, Ann; Feeser, Ingo; O'Connell, Michael; Atkinson, Tim; Jourdan, Anne-Lise; March, Anna; Fisher, Elizabeth H.

    2016-01-01

    The early Holocene cooling, which occurred around 8200 calendar years before present, was a prominent abrupt event around the north Atlantic region. Here, we investigate the timing, duration, magnitude and regional coherence of the event as expressed in carbonate oxygen-isotope records from three lakes on northwest Europe's Atlantic margin in western Ireland, namely Loch Avolla, Loch Gealáin and Lough Corrib. An abrupt negative oxygen-isotope excursion lasted about 200 years. Comparison of records from three sites suggests that the excursion was primarily the result of a reduction of the oxygen-isotope values of precipitation, which was likely caused by lowered air temperatures, possibly coupled with a change in atmospheric circulation. Comparison of records from two of the lakes (Loch Avolla and Loch Gealáin), which have differing bathymetries, further suggests a reduction in evaporative loss of lake water during the cooling episode. Comparison of climate model experiments with lake-sediment isotope data indicates that effective moisture may have increased along this part of the northeast Atlantic seaboard during the 8200-year climatic event, as lower evaporation compensated for reduced precipitation.

  12. Clinical Comparisons of Two Free Light Chain Assays to Immunofixation Electrophoresis for Detecting Monoclonal Gammopathy

    Directory of Open Access Journals (Sweden)

    Han-Sung Kim

    2014-01-01

    Full Text Available Free light chains (FLCs are useful biomarkers for the diagnosis and monitoring of various plasma cell dyscrasias. One hundred fifty-seven samples from 120 patients for screening or monitoring of monoclonal gammopathy (MG were included. The new N Latex FLC assays (Siemens Healthcare Diagnostics GmbH, Germany were compared with the Freelite FLC assays (The Binding Site Ltd., UK and the results were analyzed with those of immunofixation electrophoresis (IFE. The Freelite FLC assay showed significantly wider assay ranges than the N Latex FLC assay. The correlation coefficients of the two FLC kappa (κ assays, lambda (λ assays, and the κ/λ ratio were 0.9792, 0.8264, and 0.9064, respectively. The concordance rate was 84.7% for the FLC κ assays, 79.6% for FLC λ, and 89.2% for the κ/λ ratio. The clinical sensitivity and specificity of the κ/λ ratios were 72.2% and 93.6% for the Freelite assay and 64.6% and 100% for the N Latex FLC assay. Two FLC assays showed good correlations and concordance. However, the clinical sensitivity of the κ/λ ratio was higher in the Freelite FLC assays; clinical specificity was higher in the N Latex FLC assay. Both FLC assays seem to have limited clinical utility in detecting MG in certain clinical settings.

  13. Comparison of cell-based and PCR-based assays as methods for measuring infectivity of Tulane virus.

    Science.gov (United States)

    Shan, Lei; Yang, David; Wang, Dapeng; Tian, Peng

    2016-05-01

    In this study, we used Tulane virus (TV) as a surrogate for HuNoV to evaluate for correlation between two cell-based assays and three PCR-based assays. Specifically, the cell-based plaque and TCID50 assays measure for infectious virus particles, while the PCR-based RNase exposure, porcine gastric mucin in-situ-capture qRT-PCR (PGM-ISC-qRT-PCR), and antibody in-situ-capture qRT-PCR (Ab-ISC-qRT-PCR) assays measure for an amplicon within encapsidated viral genome. Ten batches of viral stocks ranging from 3.41 × 10(5) to 6.67 × 10(6) plaque forming units (PFUs) were used for side by side comparison with PFU as a reference. The results indicate that one PFU was equivalent to 6.69 ± 2.34 TCID50 units, 9.75 ± 10.87 RNase-untreated genomic copies (GCs), 2.87 ± 3.05 RNase-treated GCs, 0.07 ± 0.07 PGM-ISC-qRT-PCR GCs, and 0.52 ± 0.39 Ab-ISC-qRT-PCR GCs. We observed that while the cell-based assays were consistent with each other, the TCID50 assay was more sensitive than the plaque assay. In contrast, the PCR-based assays were not always consistent with the cell-based assays. The very high variations in GCs as measured by both ISC-RT-qPCR assays made them difficult to correlate against the relatively small variations (<20-fold) in the PFUs or TCID50 units as measured by the cell-based assays.

  14. Comparison of Hemoglobin A1c assay performance on two different commercial systems

    Directory of Open Access Journals (Sweden)

    Jozo Ćorić

    2015-04-01

    Full Text Available Introduction: Glycated hemoglobin (HbA1c is formed by non-enzymatic binding of glucose to the free amino group of the N-terminal end of the ß-chain of hemoglobin A. HbA1c is representative of the mean blood glucose level over three months. The aim of the study was to evaluate the Hemoglobin A1c immunoturbidimetric assay performance on two different commercial systems.Methods: We evaluated the precision and trueness for determination of HbA1c in whole blood. Concentrations of total hemoglobin and HbA1c were evaluated on Dimension Xpand (Siemens and Cobas 501 (Roche analyzers. HbA1c was measured in a latex agglutination inhibition test. Commercial controls Liquichek Diabetes Control Level 1 and Liquichek Diabetes Control Level 2 (Bio Rad at two levels were used for quality control. Analytical validation of HbA1c included: within-run imprecision, between-day imprecision, inaccuracy and comparison determination on the human samples on 2 systems: Dimension Xpand and Cobas 501 analyzers. Results: Within-run imprecision on the commercially controls for Level 1 is 4.5% and Level 2 is 3.2% between-day imprecision on commercially controls is 6.1% Level 1 and 5.1% Level 2 for respectively inac- curacy on commercially controls for Level 1 is 1.8% and Level 2 is 4.8%. Method comparison on human samples shows the correlation coefficient of 0.99.Conclusion: The presented results of the analytical evaluation methods for the determination of HbA1c showed an acceptable accuracy and precision.

  15. RAINBOW TROUT ANDROGEN RECEPTOR ALPHA AND THE HUMAN ANDROGEN RECEPTOR: COMPARISONS IN THE COS WHOLE CELL BINDING ASSAY

    Science.gov (United States)

    Rainbow Trout Androgen Receptor Alpha And Human Androgen Receptor: Comparisons in the COS Whole Cell Binding Assay Mary C. Cardon, L. Earl Gray, Jr. and Vickie S. WilsonU.S. Environmental Protection Agency, ORD, NHEERL, Reproductive Toxicology Division, Research Triangle...

  16. The rapid interphase chromosome assay (RICA implementation: comparison with other PCC methods

    Directory of Open Access Journals (Sweden)

    Sommer Sylwester

    2015-12-01

    Full Text Available A report is presented on the advantages of the rapid interphase chromosome assay (RICA and the difficulties that may be met while implementing this method for application in biological dosimetry. The RICA test can be applied on unstimulated human lymphocytes; this is an advantage in comparison with the dicentric chromosomes or micronucleus tests. In the former two tests, stimulated lymphocytes are examined and hence, 48 h more are needed to obtain cells traversing the cell cycle. Due to the use of unstimulated nondividing cells, higher numbers of cells are available for RICA analysis than for dicentric chromosomes or micronuclei tests. Moreover, the method can be applied after exposure to ionizing radiation doses in excess of 5 Gy. Such doses cause a significant cell cycle delay or result in the loss of G2 phase and mitotic cells because of apoptosis. Therefore, the traditional biodosimetry based on the evaluation of the incidence of damage to chromosomes is very difficult to carry out. This is due to the lack of an adequate number of mitotic cells for analysis. RICA is free of this disadvantage. An automatic microscope can be used to retrieve cell images; automatic image analysis can also be used.

  17. Antigenic characterisation of influenza B virus with a new microneutralisation assay: comparison to haemagglutination and sequence analysis.

    Science.gov (United States)

    Ansaldi, Filippo; Bacilieri, Sabrina; Amicizia, Daniela; Valle, Laura; Banfi, Federica; Durando, Paolo; Sticchi, Laura; Gasparini, Roberto; Icardi, Giancarlo; Crovari, Pietro

    2004-09-01

    Although the haemagglutination inhibition assay is considered the "gold standard" for antigenic characterisation of influenza viruses, some limitations of this technique are well known. A new microneutralisation assay, as a tool for antigenic characterisation of influenza B viruses, has been standardised and its performance evaluated in comparison with the haemagglutination inhibition test in the light of molecular characterisation of the haemagglutinin. Twelve B viruses belonging to the two lineages and the four sub-lineages discriminated by phylogenetic analysis of HA were tested. The microneutralisation assay clearly distinguishes viruses belonging to different lineages and, in addition, discriminates strains belonging to different sub-lineages that are poorly or not discriminated using the haemagglutination inhibition test. This new microneutralisation assay could provide a useful tool for antigenic characterisation of circulating influenza viruses and contribute, together with the haemagglutination inhibition test and sequence analysis of the haemagglutinin and neuraminidase, in the choice of the strain for use in vaccine composition.

  18. Comparison of the sensitivities of common in vitro and in vivo assays of estrogenic activity: application of chemical toxicity distributions.

    Science.gov (United States)

    Dobbins, Laura L; Brain, Richard A; Brooks, Bryan W

    2008-12-01

    A number of contaminants in municipal effluent discharges are estrogen agonists to fish. Whereas several in vitro and in vivo techniques have been developed to assess the estrogenic activity of these compounds or ambient environmental samples, previous comparisons of the relative sensitivities of these approaches remain inconclusive. We employed a probabilistic hazard assessment approach using chemical toxicity distributions (CTDs) to perform a novel evaluation of relative sensitivities of six common in vitro and in vivo assays. We predicted that there was an 8.3% (human breast ademocarcinoma cell line, MCF-7, assay), 6.3% (yeast estrogen screen assay), or 1.9% (fish hepatocyte vitellogenin, VTG, assay) probability of detecting a compound in aquatic systems that will elicit an estrogenic response at concentrations at or below 0.1 microg/L, suggesting that the MCF-7 assay was the most sensitive in vitro assay evaluated in this study. The probabilities of eliciting the estrogenic response of VTG induction at a concentration less than 0.1 microg/L in rainbow trout, fathead minnow, and Japanese medaka were determined at 29.9, 26.2, and 18.8%, respectively. Thus, rainbow trout VTG induction was the most sensitive in vivo assay assessed. Subsequently, CTDs may provide a useful technique for hazard assessment of chemical classes for which exposure data are limited and for chemicals with common toxicological mechanisms and modes of action.

  19. Comparison of Three Modelling Approaches to Simulate Regional Crop Yield: A Case Study of Winter Wheat Yield in Western Germany

    NARCIS (Netherlands)

    Soltani Largani, Afsaneh; Bakker, M.M.; Veldkamp, A.; Stoorvogel, J.J.

    2016-01-01

    The need for more comparisons among models is widely recognized. This study aimed to compare three different modelling approaches for their capability to simulate and predict trends and patterns of winter wheat yield in Western Germany. The three modelling approaches included an empirical model, a p

  20. Design, preparation and use of ligated phosphoproteins: a novel approach to study protein phosphatases by dot blot array, ELISA and Western blot assays.

    Science.gov (United States)

    Sun, Luo; Ghosh, Inca; Barshevsky, Tanya; Kochinyan, Samvel; Xu, Ming-Qun

    2007-07-01

    The study of substrate specificity of protein phosphatases (PPs) is very challenging since it is difficult to prepare a suitable phosphorylated substrate. Phosphoproteins, phosphorylated by a protein kinase, or chemically synthesized phosphopeptides are commonly used substrates for PPs. Both types of these substrates have their advantages and limitations. Phosphoproteins mimic more closely the physiologically relevant PP substrates, but their preparation is technically demanding. Synthetic phosphopeptides present advantages over proteins because they can be easily produced in large quantity and their amino acid sequence can be designed to contain potential determinants of substrate specificity. However, short peptides are less optimal compared to in vivo PP substrates and often display poor and variable binding to different matrices, resulting in low sensitivity in analysis of PP activity on solid support. In this work we utilize the intein-mediated protein ligation (IPL) technique to generate substrates for PPs, combining the advantages of proteins and synthetic peptides in one molecule. The ligation of a synthetic phosphopeptide to an intein-generated carrier protein (CP) with a one-to-one stoichiometry results in the formation of a ligated phosphoprotein (LPP). Three widely used assays, dot blot array, Western blot and ELISA were employed to study the PP activity on LPP substrates. Dephosphorylation was measured by detection of the remaining phosphorylation, or lack of it, with a phospho-specific antibody. The data show the advantage of LPPs over free peptides in assays on solid supports. LPPs exhibited enhanced binding to the matrices used in the study, which significantly improved sensitivity and consistency of the assays. In addition, saturation of the signal was circumvented by serial dilution of the assay samples. This report describes detailed experimental procedures for preparation of LPP substrates and their use in PP assays based on immobilization on

  1. International network for comparison of HIV neutralization assays: the NeutNet report.

    Directory of Open Access Journals (Sweden)

    Eva Maria Fenyö

    Full Text Available BACKGROUND: Neutralizing antibody assessments play a central role in human immunodeficiency virus type-1 (HIV-1 vaccine development but it is unclear which assay, or combination of assays, will provide reliable measures of correlates of protection. To address this, an international collaboration (NeutNet involving 18 independent participants was organized to compare different assays. METHODS: Each laboratory evaluated four neutralizing reagents (TriMab, 447-52D, 4E10, sCD4 at a given range of concentrations against a panel of 11 viruses representing a wide range of genetic subtypes and phenotypes. A total of 16 different assays were compared. The assays utilized either uncloned virus produced in peripheral blood mononuclear cells (PBMCs (virus infectivity assays, VI assays, or their Env-pseudotyped (gp160 derivatives produced in 293T cells (PSV assays from molecular clones or uncloned virus. Target cells included PBMC and genetically-engineered cell lines in either a single- or multiple-cycle infection format. Infection was quantified by using a range of assay read-outs that included extracellular or intracellular p24 antigen detection, RNA quantification and luciferase and beta-galactosidase reporter gene expression. FINDINGS: PSV assays were generally more sensitive than VI assays, but there were important differences according to the virus and inhibitor used. For example, for TriMab, the mean IC50 was always lower in PSV than in VI assays. However, with 4E10 or sCD4 some viruses were neutralized with a lower IC50 in VI assays than in the PSV assays. Inter-laboratory concordance was slightly better for PSV than for VI assays with some viruses, but for other viruses agreement between laboratories was limited and depended on both the virus and the neutralizing reagent. CONCLUSIONS: The NeutNet project demonstrated clear differences in assay sensitivity that were dependent on both the neutralizing reagent and the virus. No single assay was

  2. Preliminary report of the comparison of multiple non-destructive assay techniques on LANL Plutonium Facility waste drums

    Energy Technology Data Exchange (ETDEWEB)

    Bonner, C.; Schanfein, M.; Estep, R. [and others

    1999-03-01

    Prior to disposal, nuclear waste must be accurately characterized to identify and quantify the radioactive content. The DOE Complex faces the daunting task of measuring nuclear material with both a wide range of masses and matrices. Similarly daunting can be the selection of a non-destructive assay (NDA) technique(s) to efficiently perform the quantitative assay over the entire waste population. In fulfilling its role of a DOE Defense Programs nuclear User Facility/Technology Development Center, the Los Alamos National Laboratory Plutonium Facility recently tested three commercially built and owned, mobile nondestructive assay (NDA) systems with special nuclear materials (SNM). Two independent commercial companies financed the testing of their three mobile NDA systems at the site. Contained within a single trailer is Canberra Industries segmented gamma scanner/waste assay system (SGS/WAS) and neutron waste drum assay system (WDAS). The third system is a BNFL Instruments Inc. (formerly known as Pajarito Scientific Corporation) differential die-away imaging passive/active neutron (IPAN) counter. In an effort to increase the value of this comparison, additional NDA techniques at LANL were also used to measure these same drums. These are comprised of three tomographic gamma scanners (one mobile unit and two stationary) and one developmental differential die-away system. Although not certified standards, the authors hope that such a comparison will provide valuable data for those considering these different NDA techniques to measure their waste as well as the developers of the techniques.

  3. Evaluation of point-of-contact circulating cathodic antigen assays for the detection of Schistosoma mansoni infection in low-, moderate-, and high-prevalence schools in western Kenya.

    Science.gov (United States)

    Foo, Karen T; Blackstock, Anna J; Ochola, Elizabeth A; Matete, Daniel O; Mwinzi, Pauline N M; Montgomery, Susan P; Karanja, Diana M S; Secor, W Evan

    2015-06-01

    We evaluated the performance of a point-of-contact circulating cathodic antigen assay (POC-CCA) to detect schistosome infections in primary school children (N = 1,801) living in areas with low, moderate, and high Schistosoma mansoni prevalence in western Kenya. The commercially available assay (CCA-1) and a second, experimental formulation (CCA-2) were compared against Kato-Katz stool examinations and an anti-schistosome enzyme-linked immunosorbent assay (ELISA). A latent class model based on the four tests was used to establish "true infection status" in three different zones based on their distance from Lake Victoria. As a screening tool for community treatment according to World Health Organization (WHO) guidelines, the Kato-Katz examination was in closest agreement with the latent class model, followed by the experimental CCA-2, soluble adult worm antigen preparation (SWAP) ELISA, and CCA-1, which had high sensitivity compared with the other tests but was consistently the least specific. Our experience suggests that POC-CCA tests offer a field-friendly alternative to Kato-Katz, but need further interpretation for appropriate field use.

  4. Multicenter comparison study of both analytical and clinical performance across 4 Roche HCV RNA assays utilizing different platforms.

    Science.gov (United States)

    Vermehren, Johannes; Stelzl, Evelyn; Maasoumy, Benjamin; Michel-Treil, Veronique; Berkowski, Caterina; Marins, Ed G; Paxinos, Ellen E; Marino, Enrique; Wedemeyer, Heiner; Sarrazin, Christoph; Kessler, Harald H

    2017-01-25

    Antiviral treatment efficacy for chronic HCV infection is determined based on measurement of HCV RNA at specific time points throughout therapy by highly sensitive and accurate HCV RNA assays. This study evaluated the performance of two recently-developed real-time PCR HCV RNA assays, the cobas HCV for use on the cobas 6800/8800 systems (cobas 6800/8800 HCV) and the cobas HCV for use on the cobas 4800 system (cobas 4800 HCV) in comparison to two established assays, the COBAS AmpliPrep/COBAS TaqMan HCV Quantitative Test, version 2.0 (CAP/CTM v2) and the COBAS TaqMan HCV Test, version 2.0 for use with the High Pure System (HPS/CTM v2). Limit of detection (LOD) and linearity at lower concentrations (5-1000 IU/mL) were assessed for cobas 6800/8800 HCV and cobas 4800 HCV using WHO standard traceable panels representing HCV genotypes (GT) 1-4. Pairwise assay comparisons were also performed using 245 clinical samples representing HCV GT 1-4.cobas 6800/8800 HCV and cobas 4800 HCV were linear at low HCV RNA concentrations (<0.3 log10 IU/mL difference between expected and observed results) with LOD of 8.2 IU/mL and 11.7 IU/mL, respectively, for GT1. The new assays showed excellent agreement with CAP/CTM v2 and HPS/CTM v2 results in samples with quantifiable viral load. Concordance across the 6 million IU/mL cutoff was high among all four assays (90-94%). In conclusion, both cobas 6800/8800 HCV and cobas 4800 HCV tests are sensitive and linear, and correlate well with established Roche assays used in clinical practice.

  5. Forest response to increased disturbance in the Central Amazon and comparison to Western Amazonian forests

    Science.gov (United States)

    Holm, J. A.; Chambers, J. Q.; Collins, W.; Higuchi, N.

    2014-12-01

    Uncertainties surrounding vegetation response to increased disturbance rates associated with climate change remains a major global change issue for Amazon forests. Additionally, turnover rates in the Western Amazon are doubled compared to the Central Amazon, and notable gradients currently exist in specific wood density and aboveground biomass (AGB). This study investigates the extent to which the variation in disturbance regimes contributes to these regional gradients. To address these issues, we evaluated disturbance-recovery processes under scenarios of increased disturbance rates in a Central Amazon forest using first ZELIG-TROP, a dynamic vegetation gap model which we calibrated using long-term inventory data, and second using the Community Land Model (CLM), a global land surface model. Upon doubling the mortality rate in the Central Amazon to mirror the disturbance regime in the Western Amazon of ~2% mortality, the two regions continued to differ in multiple forest processes. With the inclusion of elevated natural disturbances, at steady-state, AGB significantly decreased by 41.9% with no significant difference between modeled AGB and empirical AGB from the western Amazon datasets (104 vs. 107 Mg C ha-1). However, different processes were responsible for the reductions in AGB between the models and empirical dataset. The empirical dataset suggests that a decrease in wood density drives the reduction in AGB. While decreased stand basal area was the driver of AGB loss in ZELIG-TROP, and decreased leaf area index (LAI) was the driver in CLM, two forest attributes that do not significantly vary across the Amazon Basin. Further comparisons found that stem density, specific wood density, and growth rates differed between the two Amazonian regions. This suggests that: 1) the variability between regions cannot be entirely explained by the variability in disturbance regime, but rather potentially sensitive to intrinsic environmental factors; or 2) the models are not

  6. Dealing with large sample sizes: comparison of a new one spot dot blot method to western blot.

    Science.gov (United States)

    Putra, Sulistyo Emantoko Dwi; Tsuprykov, Oleg; Von Websky, Karoline; Ritter, Teresa; Reichetzeder, Christoph; Hocher, Berthold

    2014-01-01

    Western blot is the gold standard method to determine individual protein expression levels. However, western blot is technically difficult to perform in large sample sizes because it is a time consuming and labor intensive process. Dot blot is often used instead when dealing with large sample sizes, but the main disadvantage of the existing dot blot techniques, is the absence of signal normalization to a housekeeping protein. In this study we established a one dot two development signals (ODTDS) dot blot method employing two different signal development systems. The first signal from the protein of interest was detected by horseradish peroxidase (HRP). The second signal, detecting the housekeeping protein, was obtained by using alkaline phosphatase (AP). Inter-assay results variations within ODTDS dot blot and western blot and intra-assay variations between both methods were low (1.04-5.71%) as assessed by coefficient of variation. ODTDS dot blot technique can be used instead of western blot when dealing with large sample sizes without a reduction in results accuracy.

  7. Performance of the NG OligoGen kit for the diagnosis of Neisseria gonorrhoeae: comparison with cobas 4800 assay.

    Science.gov (United States)

    Parra-Sánchez, M; García-Rey, S; Marcuello, A; Zakariya-Yousef, I; Bernal, S; Pueyo, I; Martín-Mazuelos, E; Palomares, J C

    2016-01-01

    PCR assays are nowadays between the most sensitive and reliable methods for screening and diagnosing sexually transmitted infections (STIs). The aim of this study was to analyze the reliability, accuracy, and usefulness of the new NG OligoGen kit in comparison with the cobas 4800 assay for the detection of Neisseria gonorrhoeae in clinical samples. A prospective study was designed for detection of N. gonorrhoeae including urine samples (n=152), rectal (n=80), endocervical (n=67), pharyngeal (n=41), and urethral swabs (n=5) that were sent from a regional STI clinic in Seville, Spain. Samples were collected from 255 (73.9%) men and 90 women. Sensitivity, specificity, positive and negative predicative values, and kappa value for N. gonorrhoeae detection using the NG OligoGen kit were 99.6%, 100%, 100%, 99.1%, and 0.99, respectively. Statistical data obtained in this study confirm the usefulness and reliable results of this new assay.

  8. Comparison of immunofluorescence and enzyme-linked immunosorbent assays for the serology of hantavirus infections.

    NARCIS (Netherlands)

    J. Groen (Jan); G. van der Groen (Guido); G. Hoofd; A.D.M.E. Osterhaus (Albert)

    1989-01-01

    textabstractThree enzyme-linked immunosorbent assay (ELISA) systems based upon different principles were developed for the serology of Hantaan virus infections and compared with an indirect immunofluorescence assay (IFA). The indirect IFA was carried out with gamma-irradiated Hantaan virus-infected

  9. Comparison of quantitative PCR assays for Escherichia coli targeting ribosomal RNA and single copy genes

    Science.gov (United States)

    Aims: Compare specificity and sensitivity of quantitative PCR (qPCR) assays targeting single and multi-copy gene regions of Escherichia coli. Methods and Results: A previously reported assay targeting the uidA gene (uidA405) was used as the basis for comparing the taxono...

  10. Comparison Research on the Chinese Traditional Art Style and Western Country's Abstract Art Style: A Theoretical Review

    Institute of Scientific and Technical Information of China (English)

    Xiaohui He

    2015-01-01

    In this paper, we conduct literature review and research on the Chinese traditional art style and western country' s abstract art style. Modernism art forms from concrete to abstract is a process of constant evolution, it is to find new pictures to present, to get rid of the bondage of traditional art concept to go on process. Any forward art could not avoid broke away from the traditional thinking, in the process of the old ideas and new concepts of conflict is inevitable. Therefore, corresponding research is urgent needed. The comparison result proves the intuition that both traditional Chinese culture and western culture based art design have their own spark points.

  11. Identification of hantavirus infection by Western blot assay and TaqMan PCR in patients hospitalized with acute kidney injury.

    Science.gov (United States)

    Oldal, Miklós; Németh, Viktória; Madai, Mónika; Kemenesi, Gábor; Dallos, Bianka; Péterfi, Zoltán; Sebők, Judit; Wittmann, István; Bányai, Krisztián; Jakab, Ferenc

    2014-06-01

    Hantaviruses, one of the causative agents of viral hemorrhagic fevers, represent a considerable healthcare threat. In Hungary, Dobrava-Belgrade virus (DOBV) and Puumala virus (PUUV) are the main circulating hantavirus species, responsible for the clinical picture known as hemorrhagic fever with renal syndrome, a disease that may be accompanied by acute kidney injury (AKI), requiring hospitalization with occasionally prolonged recovery phase. A total of 20 patient sera were collected over a 2-year period from persons hospitalized with AKI, displaying clinical signs and laboratory findings directly suggestive for hantavirus infection. Samples were tested using an immunoblot assay, based on complete viral nucleocapsid proteins to detect patients' IgM and IgG antibodies against DOBV and PUUV. In parallel, all specimens were also tested by 1-step real-time TaqMan reverse-transcriptase polymerase chain reaction to confirm infection and to determine the causative hantavirus genotype. We present here the first Hungarian clinical study spanning across 2 years and dedicated specifically to assess acute kidney injuries, in the context of hantavirus prevalence.

  12. Quantitative comparison between microfluidic and microtiter plate formats for cell-based assays.

    Science.gov (United States)

    Yin, Huabing; Pattrick, Nicola; Zhang, Xunli; Klauke, Norbert; Cordingley, Hayley C; Haswell, Steven J; Cooper, Jonathan M

    2008-01-01

    In this paper, we compare a quantitative cell-based assay measuring the intracellular Ca2+ response to the agonist uridine 5'-triphosphate in Chinese hamster ovary cells, in both microfluidic and microtiter formats. The study demonstrates that, under appropriate hydrodynamic conditions, there is an excellent agreement between traditional well-plate assays and those obtained on-chip for both suspended immobilized cells and cultured adherent cells. We also demonstrate that the on-chip assay, using adherent cells, provides the possibility of faster screening protocols with the potential for resolving subcellular information about local Ca2+ flux.

  13. Comparison of Three Nucleic Acid Amplification Assays of Cerebrospinal Fluid for Diagnosis of Cytomegalovirus Encephalitis

    Science.gov (United States)

    Bestetti, Arabella; Pierotti, Chiara; Terreni, Mariarosa; Zappa, Alessandra; Vago, Luca; Lazzarin, Adriano; Cinque, Paola

    2001-01-01

    The diagnostic reliabilities of three cytomegalovirus (CMV) nucleic acid amplification assays of cerebrospinal fluid (CSF) were compared by using CSF samples from human immunodeficiency virus-infected patients with a postmortem histopathological diagnosis of CMV encephalitis (n = 15) or other central nervous system conditions (n = 16). By using a nested PCR assay, the quantitative COBAS AMPLICOR CMV MONITOR PCR, and the NucliSens CMV pp67 nucleic acid sequence-based amplification assay, sensitivities were 93.3, 86.6, and 93.3%, respectively, and specificities were 93.7, 93.7, and 87.5%, respectively. The COBAS AMPLICOR assay revealed significantly higher CMV DNA levels in patients with diffuse ventriculoencephalitis than in patients with focal periventricular lesions. PMID:11230445

  14. Preliminary validation of assays to measure parameters of calcium metabolism in captive Asian and African elephants in western Europe.

    Science.gov (United States)

    van Sonsbeek, Gerda R; van der Kolk, Johannes H; van Leeuwen, Johannes P T M; Schaftenaar, Willem

    2011-05-01

    Hypocalcemia is a well known cause of dystocia in animals, including elephants in captivity. In order to study calcium metabolism in elephants, it is of utmost importance to use properly validated assays, as these might be prone to specific matrix effects in elephant blood. The aim of the current study was to conduct preliminary work for validation of various parameters involved in calcium metabolism in both blood and urine of captive elephants. Basal values of these parameters were compared between Asian elephants (Elephas maximus) and African elephants (Loxodonta africana). Preliminary testing of total calcium, inorganic phosphorus, and creatinine appeared valid for use in plasma and creatinine in urine in both species. Furthermore, measurements of bone alkaline phosphatase and N-terminal telopeptide of type I collagen appeared valid for use in Asian elephants. Mean heparinized plasma ionized calcium concentration and pH were not significantly affected by 3 cycles of freezing and thawing. Storage at 4 °C, room temperature, and 37 °C for 6, 12, and 24 hr did not alter the heparinized plasma ionized calcium concentration in Asian elephants. The following linear regression equation using pH (range: 6.858-7.887) and ionized calcium concentration in heparinized plasma was utilized: iCa(7.4) (mmol/l) = -2.1075 + 0.3130·pH(actual) + 0.8296·iCa(actual) (mmol/l). Mean basal values for pH and plasma in Asian elephant whole blood were 7.40 ± 0.048 and 7.49 ± 0.077, respectively. The urinary specific gravity and creatinine concentrations in both Asian and African elephants were significantly correlated and both were significantly lower in Asian elephants.

  15. RNase protection assays and RNA gel blots: a direct comparison of sensitivity.

    Science.gov (United States)

    Higgs, D C; Colbert, J T

    1992-01-01

    RNase protection assays are commonly thought to be a more sensitive means of detecting and quantitating specific mRNAs than are RNA gel blots (Northern blots). We have directly compared the sensitivity of these two approaches by assaying for known amounts of in vitro synthesized beta-glucuronidase mRNA. With the probes and protocols employed here, the ability to detect a specific mRNA was similar whether RNase protection or RNA gel blot analyses were performed.

  16. Comparison of 2 Luminex-based Multiplexed Protein Assays for Quantifying Microglia Activation and Inflammatory Proteins

    Science.gov (United States)

    2016-04-01

    streptavidin-phycoerythrin (PE) similar to sandwich enzyme-linked immunosorbent assays (ELISAs). The 3 fluorescent markers (2 beads plus PE) allow for...least expensive platform. It uses a magnetic plate to create a monolayer of beads that can be imaged with a light-emitting-diode-based imager capable... Magnetic Luminex Screening Assay Rat Premixed Multi-Analyte Kit, a kit was purchased that included all of the 17 analytes included in company’s catalog

  17. Investigation of Anti-Toxocara Antibodies in Epileptic Patients and Comparison of Two Methods: ELISA and Western Blotting

    Directory of Open Access Journals (Sweden)

    Mohammad Zibaei

    2013-01-01

    Full Text Available The relationship between Toxocara infection and epilepsy was previously demonstrated by several case-control studies and case reports. These previous studies were often based on the enzyme-linked immunosorbent assay (ELISA using Toxocara excretory-secretory antigens, which are not specific due to cross-reactivity with other parasitic infections such as ascariasis, trichuriasis, and anisakiasis. An immunoblot analysis is highly specific and can detect low levels of Toxocara antibodies. Therefore, this assay may be useful in the identification of toxocariasis in epileptic patients. We examined patients who had epilepsy and healthy subjects for seropositivity for Toxocara infection by ELISA and Western blotting. Out of 85 epileptic patients, 10 (11.8% and 3 (3.5% persons exhibited Toxocara immunoglobulin G (IgG antibodies responses by ELISA and by both techniques, respectively. Moreover, in the healthy group (, 3 (3.5% persons were positive by ELISA, but none was detected by Western blotting. This study indicates that Toxocara infection is a risk factor for epilepsy in Iran. These findings strongly suggest the need to perform Western blotting immunodiagnosis, as well as the ELISA using Toxocara excretory-secretory antigens, to improve diagnosis of human toxocariasis in patients with epilepsy.

  18. Comparison of a combined nontreponemal (VDRL) and treponemal immunoblot to traditional nontreponemal and treponemal assays.

    Science.gov (United States)

    Franken, Alicia A; Oliver, Joyce H; Litwin, Christine M

    2015-01-01

    Serology is the mainstay for the diagnosis and management of patients with syphilis. Newer technologies such as immunoblotting are now available for the diagnosis of syphilis. A commercial IgM/IgG immunoblot assay that detects both nontreponemal (VDRL-Venereal Disease Research Laboratory) and treponemal antibodies was compared with standard nontreponemal and treponemal assays. The immunoblot and T. pallidum particle agglutination assay (TP-PA) were performed on 198 samples. Ninety-seven samples were Rapid plasma reagin (RPR)-positive and one hundred one were RPR-negative. Positive RPR samples were titered by VDRL. The agreement, sensitivity, and specificity of the IgM/IgG VDRL results of the immunoblot compared to RPR were 74.2% (95% CI: 67.2-80.2), 77.3% (95% CI: 70.2-83.4), and 71.3% (95% CI: 64.4-77.1), respectively. The agreement, sensitivity, and specificity of the IgM/IgG treponemal immunoblot compared to TP-PA were 100% for all parameters, if the ten equivocal results were not used in the calculation. The treponemal portion of the ViraBlot IgM/IgG immunoblot compared well with the treponemal confirmation assay and could be a useful supplemental method to fluorescent treponemal antibody or TP-PA for the confirmation of syphilis. The addition of the detection of nontreponemal antibodies to the immunoblot assay, however, may not be of added benefit to the overall assay, due to decreased sensitivity and specificity compared to standard assays. © 2014 Wiley Periodicals, Inc.

  19. Comparison of immunochemical and radioligand binding assays for estrogen receptors in human breast tumors.

    Science.gov (United States)

    Di Fronzo, G; Miodini, P; Brivio, M; Cappelletti, V; Coradini, D; Granata, G; Ronchi, E

    1986-08-01

    We have compared a new enzyme immunoassay (EIA) for estrogen receptors (ER) with our conventional radioligand binding assays (multipoint dextran-coated charcoal assay for cytoplasmic ER and hydroxylapatite exchange assay for nuclear ER). Cytoplasmic ERs were measured in 76 human breast cancer specimens by EIA and by five-point Scatchard analysis. The correlation between the two assays yielded a straight line with a slope of 0.92 (r = 0.95; P less than 0.001); conversely, in 31 nuclear salt extracts, linear regression analysis of hydroxylapatite exchange assay data with EIA showed a clear correlation (r = 0.93; P less than 0.001) but a slope of 1.7, demonstrating that EIA detects more ER sites. The binding of the antibody to the cytoplasmic ER molecules was investigated by sucrose density gradient analysis, which showed that EIA recognizes both cytoplasmic forms (9 and 3S), but does not distinguish between them. Advantages and drawbacks of this method are discussed with respect to its application for routine receptor determination for clinical management of breast cancer patients.

  20. Inter-laboratory comparison of the in vivo comet assay including three image analysis systems.

    Science.gov (United States)

    Plappert-Helbig, Ulla; Guérard, Melanie

    2015-12-01

    To compare the extent of potential inter-laboratory variability and the influence of different comet image analysis systems, in vivo comet experiments were conducted using the genotoxicants ethyl methanesulfonate and methyl methanesulfonate. Tissue samples from the same animals were processed and analyzed-including independent slide evaluation by image analysis-in two laboratories with extensive experience in performing the comet assay. The analysis revealed low inter-laboratory experimental variability. Neither the use of different image analysis systems, nor the staining procedure of DNA (propidium iodide vs. SYBR® Gold), considerably impacted the results or sensitivity of the assay. In addition, relatively high stability of the staining intensity of propidium iodide-stained slides was found in slides that were refrigerated for over 3 months. In conclusion, following a thoroughly defined protocol and standardized routine procedures ensures that the comet assay is robust and generates comparable results between different laboratories.

  1. Comparison of a PCR assay in whole blood and serum specimens for canine brucellosis diagnosis.

    Science.gov (United States)

    Keid, L B; Soares, R M; Vasconcellos, S A; Salgado, V R; Megid, J; Richtzenhain, L J

    2010-07-17

    The performance of a serum PCR assay was compared with that of a blood PCR assay for the diagnosis of canine brucellosis caused by Brucella canis in 72 dogs. The dogs were classified into three groups (infected, non-infected and suspected brucellosis) according to the results of blood culture and serological tests. The sensitivities of blood PCR and serum PCR were, respectively, 97.14 per cent and 25.71 per cent. The specificities of both were 100 per cent. In the group of dogs with suspected brucellosis, three were positive by blood PCR and none was positive by serum PCR. Serum PCR showed little value for the direct diagnosis of canine brucellosis as the assay had low diagnostic sensitivity and fewer positive dogs were detected by this test than by blood culture, blood PCR, rapid slide agglutination test (RSAT) and RSAT with 2-mercaptoethanol.

  2. Deployable laboratory response to influenza pandemic; PCR assay field trials and comparison with reference methods.

    Directory of Open Access Journals (Sweden)

    Timothy J J Inglis

    Full Text Available BACKGROUND: The influenza A/H1N1/09 pandemic spread quickly during the Southern Hemisphere winter in 2009 and reached epidemic proportions within weeks of the official WHO alert. Vulnerable population groups included indigenous Australians and remote northern population centres visited by international travellers. At the height of the Australian epidemic a large number of troops converged on a training area in northern Australia for an international exercise, raising concerns about their potential exposure to the emerging influenza threat before, during and immediately after their arrival in the area. Influenza A/H1N1/09 became the dominant seasonal variant and returned to Australia during the Southern winter the following year. METHODS: A duplex nucleic acid amplification assay was developed within weeks of the first WHO influenza pandemic alert, demonstrated in northwestern Australia shortly afterwards and deployed as part of the pathology support for a field hospital during a military exercise during the initial epidemic surge in June 2009. RESULTS: The nucleic acid amplification assay was twice as sensitive as a point of care influenza immunoassay, as specific but a little less sensitive than the reference laboratory nucleic acid amplification assay. Repetition of the field assay with blinded clinical samples obtained during the 2010 winter influenza season demonstrated a 91.7% congruence with the reference laboratory method. CONCLUSIONS: Rapid in-house development of a deployable epidemic influenza assay allowed a flexible laboratory response, effective targeting of limited disease control resources in an austere military environment, and provided the public health laboratory service with a set of verification tools for resource-limited settings. The assay method was suitable for rapid deployment in time for the 2010 Northern winter.

  3. Comparison of three multiplex PCR assays for the detection of respiratory viral infections: evaluation of xTAG respiratory virus panel fast assay, RespiFinder 19 assay and RespiFinder SMART 22 assay

    Directory of Open Access Journals (Sweden)

    Dabisch-Ruthe Mareike

    2012-07-01

    Full Text Available Abstract Background A broad spectrum of pathogens is causative for respiratory tract infections, but symptoms are mostly similar. Therefore, the identification of the causative viruses and bacteria is only feasible using multiplex PCR or several monoplex PCR tests in parallel. Methods The analytical sensitivity of three multiplex PCR assays, RespiFinder-19, RespiFinder-SMART-22 and xTAG-Respiratory-Virus-Panel-Fast-Assay (RVP, were compared to monoplex real-time PCR with quantified standardized control material. All assays include the most common respiratory pathogens. Results To compare the analytical sensitivity of the multiplex assays, samples were inoculated with 13 different quantified viruses in the range of 101 to 105 copies/ml. Concordant results were received for rhinovirus, whereas the RVP detected influenzavirus, RSV and hMPV more frequently in low concentrations. The RespiFinder-19 and the RespiFinder-SMART-22 showed a higher analytical sensitivity for adenoviruses and coronaviruses, whereas the RVP was incapable to detect adenovirus and coronavirus in concentrations of 104 copies/ml. The RespiFinder-19 and RespiFinder-SMART-22A did not detect influenzaviruses (104 copies/ml and RSV (103 copies/ml. The detection of all 13 viruses in one sample was only achieved using monoplex PCR. To analyze possible competitive amplification reactions between the different viruses, samples were further inoculated with only 4 different viruses in one sample. Compared to the detection of 13 viruses in parallel, only a few differences were found. The incidence of respiratory viruses was compared in tracheal secretion (TS samples (n = 100 of mechanically ventilated patients in winter (n = 50 and summer (n = 50. In winter, respiratory viruses were detected in 32 TS samples (64% by RespiFinder-19, whereas the detection rate with RVP was only 22%. The most frequent viruses were adenovirus (32% and PIV-2 (20%. Multiple infections were detected

  4. GENOTOXICITY OF TEN CIGARETTE SMOKE CONDENSATES IN FOUR TEST SYSTEMS: COMPARISONS BETWEEN ASSAYS AND CONDENSATES

    Science.gov (United States)

    What is the study? This the first assessment of a set of cigarette smoke condensates from a range of cigarette types in a variety (4) of short-term genotoxicity assays. Why was it done? No such comparative study of cigarette smoke condensates has been reported. H...

  5. Comparison of Tuberculin Activity in the Interferon-gamma Assay for the Diagnosis of Bovine Tuberculosis

    Science.gov (United States)

    Cattle infected with bovine tuberculosis still represent a serious regulatory and health concern in a variety of countries. Early diagnosis using the in vitro interferon gamma (IFN-gamma) assay has been applied for more than a decade. Briefly, IFN-gamma responses in whole blood cultures stimulated w...

  6. Detection of enterovirus RNA in cerebrospinal fluid : Comparison of two molecular assays

    NARCIS (Netherlands)

    de Crom, S. C. M.; Obihara, C. C.; van Loon, A. M.; Argilagos-Alvarez, A. A.; Peeters, M. F.; van Furth, A. M.; Rossen, J. W. A.

    2012-01-01

    Enterovirus (EV) and human parechovirus (HPeV) are a major cause of infection in childhood. A rapid diagnostic test may improve the management of patients with EV and HPeV infection. The aim of this study is to evaluate the performance of the GeneXpert enterovirus assay (GXEA) for detection of EV RN

  7. COMPARISON OF AN IN VIVO FISH VTG ASSAY WITH YES AND E-SCREEN

    Science.gov (United States)

    This study compares the efficacy of two in vitro, estrogen-sensitive bioassays to rank the "relative estrogenicity" of five natural, pharmaceutical and xenoestrogens with a newly developed in vivo bioassay. The E-SCREEN (MCF-7 tumor cells) and YES (Yeast Estrogen Screen) assays w...

  8. Comparison of five commercial anti-tetanus toxoid immunoglobulin G enzyme-linked immunosorbent assays.

    Science.gov (United States)

    Perry, A L; Hayes, A J; Cox, H A; Alcock, F; Parker, A R

    2009-12-01

    Five commercially available enzyme-linked immunosorbent assays for the measurement of anti-tetanus toxoid immunoglobulin G (IgG) antibodies were evaluated for performance. The data suggest that there are manufacturer-dependent differences in sensitivity and accuracy for the determination of tetanus toxoid IgG antibodies that could result in different diagnostic interpretations.

  9. Comparison of the unlabeled and labeled pre-mRNA splicing assays in vitro

    Institute of Scientific and Technical Information of China (English)

    TIAN XU BU; JING XIN HONG; ZHI YAO; JIE YANG

    2006-01-01

    Pre-mRNA splicing is a fundamental process required for the expression of most metazoan genes. It is carried out by the spliceosome that catalyzes the removal of non-coding intron sequences to ligate exons into mature mRNA prior to transport and translation. The purpose of our study is to explore whether the in vitro unlabeled pre-mRNA splicing assay could be performed as an alternative method of splicing reaction other than the radiolabeled one. Two different splicing methods in vitro, 32P labeled and unlabeled pre-mRNA as the substrates in the reaction, were investigated. The radiolabeled products were visualized by autoradiography while the unlabeled products were observed by Ethidium Bromide (EB)staining. As a result, although there are more unspecific bands in the EB staining assay than 32P labeled one, the RNA products of in vitro splicing could be observed clearly. This suggests that the unlabeled pre-mRNA splicing assay can be an optional substitution for the isotope-labeled assay.

  10. Multicentre comparison of a diagnostic assay: Aquaporin-4 antibodies in neuromyelitis optica

    NARCIS (Netherlands)

    P. Waters (Patrick); M. Reindl (Markus); A. Saiz (Albert Abe); K. Schanda (Kathrin); F. Tuller (Friederike); V. Kral (Vlastimil); P. Nytrova (Petra); O. Sobek (Ondrej); H.H. Nielsen (Helle Hvilsted); T. Barington (Torben); S.T. Lillevang (Søren T.); Z. Illes (Zsolt); K. Rentzsch (Kristin); A. Berthele (Achim); T. Berki (Tímea); L. Granieri; A. Bertolotto (Antonio); B. Giometto; L. Zuliani (Luigi); D. Hamann (Dörte); J.L. Van Pelt (Joost L.); R.Q. Hintzen (Rogier); R. Höftberger (Romana); C. Costa (Carme); M. Comabella (Manuel); X. Montalban (Xavier); M. Tintoré; A. Siva (Aksel); A. Altintas (Ayse); G. Deniz (Gunnur); M. Woodhall (Mark); J. Palace (Jacqueline); F. Paul (Friedemann); H.P. Hartung; O. Aktas (Orhan); S. Jarius (Sven); B. Wildemann (Brigitte); C. Vedeler (Christian); A. Ruiz (Anne); M.I. Leite (M. Isabel); P. Trillenberg (Peter); M. Probst (Monika); S. Saschenbrecker (Sandra); A.J.P.E. Vincent (Arnoud); R. Marignier (Romain)

    2016-01-01

    textabstractObjective Antibodies to cell surface central nervous system proteins help to diagnose conditions which often respond to immunotherapies. The assessment of antibody assays needs to reflect their clinical utility. We report the results of a multicentre study of aquaporin (AQP) 4 antibody (

  11. Use of rapid HIV assays as supplemental tests in specimens with repeatedly reactive screening immunoassay results not confirmed by HIV-1 Western blot.

    Science.gov (United States)

    Wesolowski, Laura G; Delaney, Kevin P; Meyer, William A; Blatt, Amy J; Bennett, Berry; Chavez, Pollyanna; Granade, Timothy C; Owen, Michele

    2013-09-01

    An alternate HIV testing algorithm has been proposed which includes a fourth-generation immunoassay followed by an HIV-1/HIV-2 antibody differentiation supplemental test for reactive specimens and a nucleic acid test (NAT) for specimens with discordant results. To evaluate the performance of five rapid tests (Alere Clearview, Bio-Rad Multispot, OraSure OraQuick, MedMira Reveal, and Trinity Biotech Unigold) as the supplemental antibody assay in the algorithm. A total of 3273 serum and plasma specimens that were third-generation immunoassay repeatedly reactive and Western blot (WB) negative or indeterminate were tested with rapid tests and NAT. Specimens were classified by NAT: (1) HIV-1 infected (NAT-reactive; n=184, 5.6%), (2) HIV-status unknown (NAT nonreactive; n=3078, 94.2%) or by Multispot, (3) HIV-2 positive (n=5), and (4) HIV-1 and HIV-2 positive (n=6). Excluding HIV-2 positive specimens, we calculated the proportion of reactive rapid tests among specimens with reactive and nonreactive NAT. The proportion of infected specimens with reactive rapid test results and negative or indeterminate WB ranged from 30.4% (56) to 47.8% (88) depending on the rapid test. From 1% to 2% of NAT-negative specimens had reactive rapid test results. In these diagnostically challenging specimens, all rapid tests identified infections that were missed by the Western blot, but only Multispot could differentiate HIV-1 from HIV-2. Regardless of which rapid test is used as a supplemental test in the alternative algorithm, false-positive algorithm results (i.e., reactive screening and rapid test in uninfected person) may occur, which will need to be resolved during the baseline medical evaluation. Published by Elsevier B.V.

  12. Comparison of microscopy and Alamar blue reduction in a larval based assay for schistosome drug screening.

    Directory of Open Access Journals (Sweden)

    Nuha R Mansour

    Full Text Available BACKGROUND: In view of the current widespread use of and reliance on a single schistosomicide, praziquantel, there is a pressing need to discover and develop alternative drugs for schistosomiasis. One approach to this is to develop High Throughput in vitro whole organism screens (HTS to identify hits amongst large compound libraries. METHODOLOGY/PRINCIPAL FINDINGS: We have been carrying out low throughput (24-well plate in vitro testing based on microscopic evaluation of killing of ex-vivo adult S. mansoni worms using selected compound collections mainly provided through the WHO-TDR Helminth Drug Initiative. To increase throughput, we introduced a similar but higher throughput 96-well primary in vitro assay using the schistosomula stage which can be readily produced in vitro in large quantities. In addition to morphological readout of viability we have investigated using fluorometric determination of the reduction of Alamar blue (AB, a redox indicator of enzyme activity widely used in whole organism screening. A panel of 7 known schistosome active compounds including praziquantel, produced diverse effects on larval morphology within 3 days of culture although only two induced marked larval death within 7 days. The AB assay was very effective in detecting these lethal compounds but proved more inconsistent in detecting compounds which damaged but did not kill. The utility of the AB assay in detecting compounds which cause severe morbidity and/or death of schistosomula was confirmed in testing a panel of compounds previously selected in library screening as having activity against the adult worms. Furthermore, in prospective library screening, the AB assay was able to detect all compounds which induced killing and also the majority of compounds designated as hits based on morphological changes. CONCLUSION: We conclude that an HTS combining AB readout and image-based analysis would provide an efficient and stringent primary assay for schistosome

  13. Comparison of static and microfluidic protease assays using modified bioluminescence resonance energy transfer chemistry.

    Directory of Open Access Journals (Sweden)

    Nan Wu

    Full Text Available BACKGROUND: Fluorescence and bioluminescence resonance energy transfer (F/BRET are two forms of Förster resonance energy transfer, which can be used for optical transduction of biosensors. BRET has several advantages over fluorescence-based technologies because it does not require an external light source. There would be benefits in combining BRET transduction with microfluidics but the low luminance of BRET has made this challenging until now. METHODOLOGY: We used a thrombin bioprobe based on a form of BRET (BRET(H, which uses the BRET(1 substrate, native coelenterazine, with the typical BRET(2 donor and acceptor proteins linked by a thrombin target peptide. The microfluidic assay was carried out in a Y-shaped microfluidic network. The dependence of the BRET(H ratio on the measurement location, flow rate and bioprobe concentration was quantified. Results were compared with the same bioprobe in a static microwell plate assay. PRINCIPAL FINDINGS: The BRET(H thrombin bioprobe has a lower limit of detection (LOD than previously reported for the equivalent BRET(1-based version but it is substantially brighter than the BRET(2 version. The normalised BRET(H ratio of the bioprobe changed 32% following complete cleavage by thrombin and 31% in the microfluidic format. The LOD for thrombin in the microfluidic format was 27 pM, compared with an LOD of 310 pM, using the same bioprobe in a static microwell assay, and two orders of magnitude lower than reported for other microfluidic chip-based protease assays. CONCLUSIONS: These data demonstrate that BRET based microfluidic assays are feasible and that BRET(H provides a useful test bed for optimising BRET-based microfluidics. This approach may be convenient for a wide range of applications requiring sensitive detection and/or quantification of chemical or biological analytes.

  14. Optimizing the HRP-2 In Vitro Malaria Drug Susceptibility Assay Using a Reference Clone to Improve Comparisons of Plasmodium falciparum Field Isolates

    Science.gov (United States)

    2012-09-13

    available soon. Optimizing the HRP-2 in vitro malaria drug susceptibility assay using a reference clone to improve comparisons of Plasmodium falciparum...Optimizing the HRP-2 in vitro malaria drug susceptibility assay using a reference clone to improve comparisons of Plasmodium falciparum field isolates 5a...Date: 13 September 2012 14. ABSTRACT Apparent emerging artemisinin-resistant Plasmodium falciparum malaria in Southeast Asia requires development

  15. Biogeographic comparison of Lophelia-associated bacterial communities in the Western Atlantic reveals conserved core microbiome

    Science.gov (United States)

    Kellogg, Christina A.; Goldsmith, Dawn; Gray, Michael A.

    2017-01-01

    Over the last decade, publications on deep-sea corals have tripled. Most attention has been paid to Lophelia pertusa, a globally distributed scleractinian coral that creates critical three-dimensional habitat in the deep ocean. The bacterial community associated with L. pertusa has been previously described by a number of studies at sites in the Mediterranean Sea, Norwegian fjords, off Great Britain, and in the Gulf of Mexico (GOM). However, use of different methodologies prevents direct comparisons in most cases. Our objectives were to address intra-regional variation and to identify any conserved bacterial core community. We collected samples from three distinct colonies of L. pertusa at each of four locations within the western Atlantic: three sites within the GOM and one off the east coast of the United States. Amplicon libraries of 16S rRNA genes were generated using primers targeting the V4–V5 hypervariable region and 454 pyrosequencing. The dominant phylum was Proteobacteria (75–96%). At the family level, 80–95% of each sample was comprised of five groups: Pirellulaceae, Pseudonocardiaceae, Rhodobacteraceae, Sphingomonadaceae, and unclassified Oceanospirillales. Principal coordinate analysis based on weighted UniFrac distances showed a clear distinction between the GOM and Atlantic samples. Interestingly, the replicate samples from each location did not always cluster together, indicating there is not a strong site-specific influence. The core bacterial community, conserved in 100% of the samples, was dominated by the operational taxonomic units of genera Novosphingobium and Pseudonocardia, both known degraders of aromatic hydrocarbons. The sequence of another core member, Propionibacterium, was also found in prior studies of L. pertusa from Norway and Great Britain, suggesting a role as a conserved symbiont. By examining more than 40,000 sequences per sample, we found that GOM samples were dominated by the identified conserved core sequences, whereas

  16. Comparison of the Freelite serum free light chain (SFLC) assay with serum and urine electrophoresis/immunofixation and the N Latex FLC assay.

    Science.gov (United States)

    Sasson, S C; McGill, K; Wienholt, L; Carr, A; Brown, D A; Kelleher, A D; Breit, S N; Sewell, W A

    2015-10-01

    Few reports have compared available serum free light chain (SFLC) assays. Here, a retrospective audit of the Freelite SFLC assay compared results to electrophoresis (EP)/immunofixation (IFX) and the N Latex FLC assay.A total of 244 samples collected over 3.5 months were studied using the Freelite and N Latex FLC nephelometry assays. Results were compared with serum and/or urine EP/IFX. The precision and linearity of the N Latex FLC assay was examined.Detectable paraprotein by serum or urine EP/IFX was present in 94% of samples with kappa and 100% with lambda FLC restriction. The correlation between the assays was higher for kappa (rho = 0.97) than lambda (rho = 0.89) especially when lambda results were above the upper limit of normal (rho = 0.62). Agreement in the categorical diagnosis as measured by the Cohen's kappa statistic was good (0.70). The N Latex FLC assay displayed good precision and linearity. In discordant samples the Freelite and N Latex FLC assays had equivalent agreement with IFX.Traditional methods of EP/IFX detected paraproteins in the majority of cases. Correlation between the Freelite and N Latex FLC assay is better for kappa than lambda FLC. The two assays are not entirely equivalent. Care should be taken by interpreting physicians and laboratories considering switching assays.

  17. A comparison between radioligand and immunohistochemical assay of hormone receptors in primary breast cancer.

    Science.gov (United States)

    Charalambous, D; Kitchen, P R; Stillwell, R G; Smart, P J; Rode, J

    1993-08-01

    The detection of oestrogen and progesterone receptor (ER and PgR) levels in human breast carcinoma has traditionally been performed using a biochemical radioligand binding method. This method has several disadvantages including the requirement for generous tissue samples, the production of radioactive waste products and the inability to exclude non-malignant cellular material from the assay process. An alternative method for detecting hormone receptors is available with the use of a monoclonal antibody specific for the ER or PgR receptor using immunocytochemical assay (ER-ICA or PgR-ICA). Although designed for use on frozen section material, with modifications this method can be used on paraffin sections of routinely fixed and processed tissue, on archival material and on very small specimens. Further, an objective assessment or scoring of staining intensity is possible using computerized video-image analysis. Forty-three cases of primary breast carcinoma, treated from 1989 to 1991 at Goulburn Valley Base Hospital, Shepparton were assessed for ER and PgR content using both the radioligand method and immunohistochemistry with video-image analysis, and the results were compared. Of the 43 cases, ER-ICA and ER had a concordance of 81% (P < 0.001, r = 0.58) and in 39 cases, PgR and PgR-ICA had a concordance of 87% (P < 0.001, r = 0.54). Because the sample for radioligand assay is of uncertain composition and the immunohistochemical stain can be scored specifically for malignant epithelium, a degree of discordance is thought to be mostly attributable to the limitations of the radioligand assay.

  18. Immunoradiometric assay for prolactin in serum and tissue; Comparison with radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Ohnami, Shumpei; Nakata, Hajime; Eto, Sumiya (University of Occupational and Environmental Health Hospital, Kitakyushu, Fukuoka (Japan))

    1990-09-01

    Prolactin (PRL) concentrations in sera and tumors of patients with various pituitary tumors were measured by both immunoradiometric assay (IRMA) and radioimmunoassay (RIA). PRL concentrations in sera and tumor tissues measured by IRMA were well correlated with those measured by RIA. PRL concentrations in sera reflected those of tumors removed. This IRMA is a simple and useful method for PRL determination in serum and tissue. (author).

  19. Comparison of three cyanogen assays for total cyanogens in cassava (Manihot esculenta Crantz)

    DEFF Research Database (Denmark)

    Saka, J.D.K.; Mhone, A.R.K.; Brimer, Leon

    1997-01-01

    The sensitivity and reproducibility of three methods for determining the total cyanogenic potential (CNp) of 7 fresh and processed cassava varieties were determined and compared. The total cyanogen content of fresh cassava roots and three cassava products (kondowole, makaka, and starch) were...... analysed by the acid hydrolysis, microdiffusion with solid state detection and Cooke's enzymatic assays. The total cyanogen contents of the cassava, obtained by the three methods were not significantly different (p

  20. Determinants of the sustainability of farming on leased agricultural land in Poland in comparison with selected Western European countries

    Directory of Open Access Journals (Sweden)

    Adam Majchrzak

    2014-08-01

    Full Text Available In Western Europe the lease is the dominant form of land management in agriculture. This is not only due to the tradition of this institution, but also to the scope of the regulations that guarantee the stability of the legal relationship and ensure the independence of the lessee in production process based on the subject of the lease. This article attempts to assess the instruments for the protection of farming on leased agricultural land in Poland in comparison with solutions applicable in after countries of Western Europe. In this paper, a review of regulations relating to the length of the lease, the obligation to care for the preservation of the quality of the resource, as well as the criteria for determining the amount of the rent, is examines . On this basis, it has been shown that the regulations determining the stability of the lease in Poland deviate far from the norms provided in most Western European countries. This implies that the legal position of a lessee in Poland is weaker than it is in the rest of Western Europe, which is a negative assessment from point of view the of agrarian structure in Poland.

  1. [Comparison between transmission of modern western medicine in China and Japan].

    Science.gov (United States)

    Liu, Yuan-Ming

    2009-05-01

    In the middle period of the 16th century, western medicine had been introduced successively into China and Japan by the medium of Catholic missionaries. The transmission mode of western medicine in the two countries differed dramatically from each other due to the political, economic and social cultural differences of that time. In China, the transmission of western medicine focused on the theory first and transferred to the practical use later; while in Japan, it began with clinical treatment, then rose from the technology to the theory. As a result, the cognition to the western medical knowledge and medical system as well as the transplanting and localization in China and Japan were significantly different. The local western medical groups emerged earlier in Japan than in China, and the attitude toward western medicine was also more positive and a national health system was quickly established. The corresponding situation in China obviously lagged behind the one in Japan, and China learned from the successful experiences of Japan for a time.

  2. Comparison of various assays to quantitate macrophage activation by biological response modifiers

    Energy Technology Data Exchange (ETDEWEB)

    Schultz, R.M.; Nanda, S.; Altom, M.G.

    1984-01-01

    Macrophages treated with various compounds that enhance host antitumor resistance exhibit measurable changes in metabolism, function, and surface antigens. In this study, murine peptone-induced peritoneal macrophages were stimulated in vitro by bacterial lipopolysaccharide (LPS), muramyl dipeptide (MDP), and poly I.poly C. They were subsequently compared in their ability to release superoxide and act as tumoristatic and tumoricidal effector cells. Superoxide generation was assayed by the reduction of ferricytochrome C. All three compounds failed to induce significant O/sub 2/- release, unless the cells were also treated with phorbol myristate acetate (PMA). MDP was most active in potentiating the PMA response. In the tumor growth inhibition assay, cytostatic activity was comparable for all three compounds and did not exceed 32 percent. The combination of subthreshold levels of these compounds and hybridoma-derived MAF acted synergistically to induce potent cytostatic activity. In the chromium release assay, LPS and poly I.poly C rendered macrophages cytolytic for P815 target cells at concentrations greater than or equal to 1 microgram/ml. In contrast, significant cytolysis was observed with MDP only at 100 micrograms/ml. Defining precisely the effect of various biological response modifiers on several parameters of macrophage function may facilitate use of these agents in cancer therapy.

  3. Comparison of mRNA Splicing Assay Protocols across Multiple Laboratories

    DEFF Research Database (Denmark)

    Whiley, Phillip; de la Hoya, Miguel; Thomassen, Mads

    2014-01-01

    and differences in results derived from analysis of a panel of breast cancer 1, early onset (BRCA1) and breast cancer 2, early onset (BRCA2) gene variants known to alter splicing (BRCA1: c.135-1G>T, c.591C>T, c.594-2A>C, c.671-2A>G, and c.5467+5G>C and BRCA2: c.426-12_8delGTTTT, c.7988A>T, c.8632+1G>A, and c.9501...... in turn relies on appropriate assay design, interpretation, and reporting.METHODS: We conducted a multicenter investigation to compare mRNA splicing assay protocols used by members of the ENIGMA (Evidence-Based Network for the Interpretation of Germline Mutant Alleles) consortium. We compared similarities...... for accurate splicing assay results. For example, because of the position of primers and PCR extension times, several isoforms associated with BRCA1, c.594-2A>C and c.671-2A>G, were not detected by many sites. Variation was most evident for the detection of low-abundance transcripts (e.g., BRCA2 c.8632+1G>A Δ...

  4. Comparison of microbial community assays for the assessment of stream biofilm ecology.

    Science.gov (United States)

    Vinten, A J A; Artz, R R E; Thomas, N; Potts, J M; Avery, L; Langan, S J; Watson, H; Cook, Y; Taylor, C; Abel, C; Reid, E; Singh, B K

    2011-06-01

    We investigated a range of microbiological community assays performed on scrapes of biofilms formed on artificial diffusing substrates deployed in 8 streams in eastern Scotland, with a view to using them to characterize ecological response to stream water quality. The assays considered were: Multiplex Terminal Restriction Fragment Length Polymorphism or M-TRFLP (a molecular method), Phospholipid Fatty Acid or PLFA analysis (a biochemical method) and MICRORESP™ (a physiological method) alongside TDI, diatom species, and chlorophyll a content. Four of the streams were classified as of excellent status (3-6μg/L Soluble Reactive Phosphorus (SRP)) with respect to soluble P content under the EU Water Framework Directive and four were of borderline good/moderate or moderate status (43-577μg/L SRP). At each site, 3 replicates of 3 solute diffusion treatments were deployed in a Latin square design. Solute diffusion treatments were: KCl (as a control solute), N and P (to investigate the effect of nutrient enrichment), or the herbicide isoproturon (as a "high impact" control, which aimed to affect biofilm growth in a way detectable by all assays). Biofilms were sampled after 4weeks deployment in a low flow period of early summer 2006. The chlorophyll a content of biofilms after 4weeks was 2.0±0.29mg/m(2) (mean±se). Dry matter content was 16.0±13.1g/m(2). The M-TRFLP was successfully used for generating community profiles of cyanobacteria, algae and bacteria and was much faster than diatom identification. The PFLA and TDI were successful after an increase in the sample size, due to low counts. The MICRORESP(™) assays were often below or near detection limit. We estimated the per-sample times for the successful assays as follows: M-TRFLP: 20min, PLFA 40min, TDI 90min. Using MANOVA on the first 5 principal co-ordinates, all the assays except MICRORESP(™) showed significant differences between sites, but none of the assays showed a significant effect of either initial

  5. [Comparison of the clinical performance of the ECLusys HIV combi assay with the Lumipulse f and HISCL 2000-i HIV-1/2 ab screening assays].

    Science.gov (United States)

    Sugiura, Aya; Iwahara, Kunihiro; Suga, Yasuyuki; Uchiyama, Sachinori; Maekawa, Masato

    2012-04-01

    We compared the ECLusys HIV combi assay (ECL HIV Ag/Ab) to the Lumipulse Forte (LPf HIV 1/2 Ab) and HISCL (HIS HIV 1/2 Ab) assays. In a dilution sensitivity test using dilution panels of WHO HIV antibody international reference panel (HIV-1 Subtype A, B, C, E, HIV-1 Group O, HIV-2) and HIV-1/2 Ab CE marked material(HIV-1, HIV-2) parent specimens, the ECL assay enabled detection at a higher level of sensitivity than either the LPf assay or the HIS assay for all dilution panels. In an early detection test in the early phase of infection in which a BBI HIV seroconversion panel was used, the ECL assay enabled detection 7 days after initial blood sample collection, whereas the LPf and HIS assays enabled detection after 27 days. In a specificity test using high RF positive specimens (n=33), pregnancy specimens (n=35), cytomegalovirus antibody positive specimens (n=36), and high M protein positive specimens (n=21) that were confirmed negative for HIV-1/2 antibodies by the LPf assay, negative results were obtained for all specimens on both the ECL assay and the HIS assay. In a correlation test using routinely collected clinical specimens (n=121), including positive stock specimens, the ECL and HIS assays demonstrated the highest agreement rate 98.3%. The above results confirmed that the fourth-generation reagent ECL assay, which simultaneously detects both HIV-1/2 antibodies and p24 antigens, is both highly sensitive and specific, and is a suitable assay for use in routine testing.

  6. Biofilm removal by medical device cleaners: comparison of two bioreactor detection assays.

    Science.gov (United States)

    Hadi, R; Vickery, K; Deva, A; Charlton, T

    2010-02-01

    Currently there are no standards for testing efficacy of medical device cleaners. With fears of prion transmission, residual protein on medical devices needs to be minimised. A bioreactor model was used to grow Pseudomonas aeruginosa biofilm on polytetrafluoroethylene coupons. The biofilm was subjected to various cleaners and residual biofilm was detected either by Crystal Violet assay (CrV) or a commercially available protein assay (PA) following hydrolysis of the biofilm. Percentage reduction of biofilm was compared with untreated controls in three independent tests. There was no significant difference in percentage biofilm reduction irrespective of whether the CrV or PA was used to detect residual biofilm. Processing of coupons attached to the bioreactor rod and position of coupon within the rod had no significant effect on cleaning efficiency or detection of residual biofilm. Both within-run and between-run variation was very low for good cleaners such as 10g/L NaOH, Zen, and 3M Rapid Multi-Enzyme Cleaner (RMEC) 70500 but was higher for poor cleaners such as Tween 20 which removed less than 20% of the biofilm. Confocal microscopy and electron microscopy provided visual confirmation of the assay results. We propose that this method is suitable as a test method for evaluating the efficacy of surgical instrument cleaners in removing biofilm, as both within-run and between-run variation was low, detection of residual biofilm can be done using either CrV or PA, and the apparatus is easy to use, cheap and readily available.

  7. Comparison of three cell fixation methods for high content analysis assays utilizing quantum dots.

    Science.gov (United States)

    Williams, Y; Byrne, S; Bashir, M; Davies, A; Whelan, A; Gun'ko, Y; Kelleher, D; Volkov, Y

    2008-10-01

    Semiconductor nanoparticles or quantum dots are being increasingly utilized as fluorescent probes in cell biology both in live and fixed cell assays. Quantum dots possess an immense potential for use in multiplexing assays that can be run on high content screening analysers. Depending on the nature of the biological target under investigation, experiments are frequently required on cells retaining an intact cell membrane or also on those that have been fixed and permeabilized to expose intracellular antigens. Fixation of cell lines before or after the addition of quantum dots may affect their localization, emission properties and stability. Using a high content analysis platform we perform a quantitative comparative analysis of three common fixation techniques in two different cell lines exposed to carboxylic acid stabilized CdTe quantum dots. Our study demonstrates that in prefixed and permeabilized cells, quantum dots are readily internalized regardless of cell type, and their intracellular location is primarily determined by the properties of the quantum dots themselves. However, if the fixation procedures are preformed on live cells previously incubated with quantum dots, other important factors have to be considered. The choice of the fixative significantly influences the fluorescent characteristics of the quantum dots. Fixatives, regardless of their chemical nature, negatively affected quantum dots fluorescence intensity. Comparative analysis of gluteraldehyde, methanol and paraformaldehyde demonstrated that 2% paraformaldehyde was the fixative of choice. The presence of protein in the media did not significantly alter the quantum dot fluorescence. This study indicates that multiplexing assays utilizing quantum dots, despite being a cutting edge tool for high content cell imaging, still require careful consideration of the basic steps in biological sample processing.

  8. Comparison of functional assays used in the clinical development of a placental malaria vaccine

    DEFF Research Database (Denmark)

    Pehrson, Caroline; Heno, Kristine K; Adams, Yvonne;

    2017-01-01

    are in clinical development. The purpose of this study was to evaluate the robustness and comparability of binding inhibition assays used in the clinical development of placental malaria vaccines. METHODS: The ability of sera from animals immunised with different VAR2CSA constructs to inhibit IE binding to CSA......BACKGROUND: Malaria in pregnancy is associated with significant morbidity in pregnant women and their offspring. Plasmodium falciparum infected erythrocytes (IE) express VAR2CSA that mediates binding to chondroitin sulphate A (CSA) in the placenta. Two VAR2CSA-based vaccines for placental malaria...

  9. Outliers affecting cardiac troponin I measurement: comparison of a new high sensitivity assay with a contemporary assay on the Abbott ARCHITECT analyser.

    Science.gov (United States)

    Sawyer, Nicola; Blennerhassett, John; Lambert, Ramon; Sheehan, Paul; Vasikaran, Samuel D

    2014-07-01

    False-positive cardiac troponin (Tn) results caused by outliers have been reported on various analytical platforms. We have compared the precision profile and outlier rate of the Abbott Diagnostics contemporary troponin I (TnI) assay with their high sensitivity (hs) TnI assay. Three studies were conducted over a 10-month period using routine patients' samples. TnI was measured in duplicate using the contemporary TnI assay in Study 1 and Study 2 (n = 7011 and 7089) and the hs-TnI assay in Study 3 (n = 1522). Critical outliers were defined as duplicate results whose absolute difference exceeded a critical difference (CD = z x √2 x SDAnalytical) at a probability level of 0.0005, with one of the results on the opposite side of the decision limit to its partner. The TnI concentration at 10% imprecision (coefficient of variation) for the contemporary TnI assay was 0.034 µg/L (Study 1) and 0.042 µg/L (Study 2), and 0.006 µg/L (6 ng/L) for the hs-TnI assay. The critical outlier rates for the contemporary TnI assay were 0.51% (Study 1) and 0.37% (Study 2) using a cut-off of 0.04 µg/L, and 0% for the hs-TnI assay using gender-specific cut-offs. The significant number of critical outliers detected using the contemporary TnI assay may pose a risk for misclassification of patients. By contrast, no critical outliers were detected using the hs-TnI assay. However, the total outlier rates for both assays were significantly higher than the expected variability of either assay. The cause of these outliers remains unclear. © The Author(s) 2013 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.

  10. Accuracy of the Fluorescence-Activated Cell Sorting Assay for the Aquaporin-4 Antibody (AQP4-Ab): Comparison with the Commercial AQP4-Ab Assay Kit

    Science.gov (United States)

    Kim, Yoo-Jin; Cheon, So Young; Kim, Boram; Jung, Kyeong Cheon; Park, Kyung Seok

    2016-01-01

    Background The aquaporin-4 antibody (AQP4-Ab) is a disease-specific autoantibody to neuromyelitis optica (NMO). We aimed to evaluate the accuracy of the FACS assay in detecting the AQP4-Ab compared with the commercial cell-based assay (C-CBA) kit. Methods Human embryonic kidney-293 cells were transfected with human aquaporin-4 (M23) cDNA. The optimal cut off values of FACS assay was tested using 1123 serum samples from patients with clinically definite NMO, those at high risk for NMO, patients with multiple sclerosis, patients with other idiopathic inflammatory demyelinating diseases, and negative controls. The accuracy of FACS assay and C-CBA were compared in consecutive 225 samples that were collected between January 2014 and June 2014. Results With a cut-off value of MFIi of 3.5 and MFIr of 2.0, the receiver operating characteristic curve for the FACS assay showed an area under the curve of 0.876. Among 225 consecutive sera, the FACS assay and C-CBA had a sensitivity of 77.3% and 69.7%, respectively, in differentiating the sera of definite NMO patients from sera of controls without IDD or of MS. Both assay had a good specificity of 100% in it. The overall positivity of the C-CBA among FACS-positive sera was 81.5%; moreover, its positivity was low as 50% among FACS-positive sera with relatively low MFIis. Conclusions Both the FACS assay and C-CBA are sensitive and highly specific assays in detecting AQP4-Ab. However, in some sera with relatively low antibody titer, FACS-assay can be a more sensitive assay option. In real practice, complementary use of FACS assay and C-CBA will benefit the diagnosis of NMO patients, because the former can be more sensitive among low titer sera and the latter are easier to use therefore can be widely used. PMID:27658059

  11. [Comparison of the clinical performance of the ECLusys HBsAg II assay with the Lumipulse f and HISCL 2000-i HBsAg screening assays].

    Science.gov (United States)

    Sugiura, Aya; Iwahara, Kunihiro; Suga, Yasuyuki; Uchiyama, Sachinori; Maekawa, Masato

    2012-02-01

    We compared the ECLusys HBsAgII (ECL HBsAg) assay to the Lumipulse Forte (LPf HBsAg) and HISCL (HIS HBsAg) assays. Measurement of dilution panels for which the WHO HBsAg international reference panel was the parent specimen revealed that the ECL and HIS assays enabled detection to a theoretical level of 0.04 IU/mL, whereas the LPf assay enabled detection to a level of 0.08 IU/mL. In a specificity test using high RF positive specimens (n = 33), pregnancy specimens (n = 35), cytomegalovirus antibody positive specimens (n = 36), and high M protein positive specimens (n = 21) that were confirmed negative for HBsAg by the LPf assay, negative results were obtained for all specimens on the HIS assay, but the ECL assay yielded a positive result for one of the high RF positive specimens. This individual was suggested on further testing to be an HBV carrier who was strongly positive for HBc antibody. In HBsAg mutants detection test, the detection rate was 92.3% with the ECL assay and 69.2% with the HIS assay. In a correlation test using routinely collected clinical specimens (n = 155), including positive stock specimens, aside from the one case where the LPf assay gave a negative result but both the ECL and HIS assays gave positive results, all of the results were consistent for all specimens. The above results confirmed that the ECL assay is both highly sensitive and specific, and also enables a high rate of HBsAg mutant detection.

  12. Comparison of a Multiplex Flow Cytometric Assay with Enzyme-Linked Immunosorbent Assay for Quantitation of Antibodies to Tetanus, Diphtheria, and Haemophilus influenzae Type b

    OpenAIRE

    Pickering, Jerry W.; Martins, Thomas B.; Schroder, M. Carl; Hill, Harry R.

    2002-01-01

    We developed a multiplexed indirect immunofluorescence assay for antibodies to Haemophilus influenza type b (Hib) polysaccharide and the toxoids of Clostridium tetani (Tet) and Corynebacterium diphtheriae (Dip) based on the Luminex multiple-analyte profiling system. A pooled serum standard was calibrated against World Health Organization standards for Dip and Tet and an international standard for Hib. The multiplexed Luminex assay was compared to individual enzyme-linked immunosorbent assays ...

  13. Inter-laboratory and inter-assay comparison on two real-time PCR techniques for quantification of PCV2 nucleic acid extracted from field samples

    DEFF Research Database (Denmark)

    Hjulsager, Charlotte Kristiane; Grau-Roma, L.; Sibila, M.

    2009-01-01

    ) diagnosis has been suggested. However, neither inter-laboratory nor inter-assay comparisons have been published so far. In the present study two different qPCR probe assays Used routinely in two laboratories were compared on DNA extracted From serum, nasal and rectal swabs. Results showed a significant......Several real-time PCR assays for quantification of PCV2 DNA (qPCR) have been described in the literature. and different in-house assays are being used by laboratories around the world. A general threshold of it copies of PCV2 per millilitre serum for postweaning multisystemic wasting syndrome (PMWS...

  14. Comparison of acrylamide intake from Western and guideline based diets using probabilistic techniques and linear programming.

    Science.gov (United States)

    Katz, Josh M; Winter, Carl K; Buttrey, Samuel E; Fadel, James G

    2012-03-01

    Western and guideline based diets were compared to determine if dietary improvements resulting from following dietary guidelines reduce acrylamide intake. Acrylamide forms in heat treated foods and is a human neurotoxin and animal carcinogen. Acrylamide intake from the Western diet was estimated with probabilistic techniques using teenage (13-19 years) National Health and Nutrition Examination Survey (NHANES) food consumption estimates combined with FDA data on the levels of acrylamide in a large number of foods. Guideline based diets were derived from NHANES data using linear programming techniques to comport to recommendations from the Dietary Guidelines for Americans, 2005. Whereas the guideline based diets were more properly balanced and rich in consumption of fruits, vegetables, and other dietary components than the Western diets, acrylamide intake (mean±SE) was significantly greater (Plinear programming and results demonstrate that linear programming techniques can be used to model specific diets for the assessment of toxicological and nutritional dietary components.

  15. [Comparison of five commercial assays for the detection of Legionella pneumophila antigens in urine].

    Science.gov (United States)

    de Ory, Fernando; Minguito, Teodora

    2009-02-01

    Antigenuria detection is the main approach for diagnosing Legionella infections. The aim of this study was to compare 5 commercially available methods for detecting Legionella pneumophila soluble antigens in urine. Seventy-one urine samples were tested, 62 from patients with bacterial infection and 9 from patients with respiratory syncytial virus infection. All samples were assayed for the presence of L. pneumophila by immunoenzymatic (ELISA) (Binax and Bartels), and immunochromatographic (IC) (Binax, SAS and Uni-Gold) methods. Identical results (35 positive and 17 negative) were obtained by the 5 assays in 52 samples (73.2%). Samples showing discrepant results were classified by the majority criterion, and/or other laboratory results (serology), and/or epidemiological findings. On this basis, 51 samples were ultimately classified as positive, and 20 as negative. Sensitivity values of ELISA-Binax, ELISA-Bartels, IC-Binax, IC-SAS and IC-Uni-Gold were 80.4, 100, 82.4, 86.3, and 70.6%, respectively. Corresponding values for specificity were 90, 95, 100, 95 and 100%. The results indicate that the methods compared are all adequate for diagnosing Legionella infection, although some have certain limitations regarding sensitivity.

  16. Comparison of Candidate Pairs of Hydrolytic Enzymes for Spectrophotometric-dual-enzyme-simultaneous-assay.

    Science.gov (United States)

    Liu, Hongbo; Yuan, Mei; Yang, Xiaolan; Hu, Xiaolei; Liao, Juan; Dang, Jizheng; Xie, Yanling; Pu, Jun; Li, Yuanli; Zhan, Chang-Guo; Liao, Fei

    2015-01-01

    Spectrophotometric-dual-enzyme-simultaneous-assay (SDESA) for enzyme-linked-immunosorbent-assay (ELISA) of two components in one well is a patented platform when a special pair of labels is accessible. With microplate readers, alkaline phosphatase on 4-nitro-1-naphthylphosphate (4NNPP) served as label A; Pseudomonas aeruginosa arylsulfatase (PAAS) and acetylcholinesterase (AChE) on their substrates derived from 4-nitrophenol/analogue served as candidate label B, and were compared for SDESA with an engineered alkaline phosphatase of Eschrichia coli (ECAP). For SDESA, the interference from overlapped absorbance was corrected based on linear additivity of absorbance to derive initial rates reflected by absorbance change at 450 nm for ECAP and at 405 nm for PAAS or AChE, after the correction of spontaneous hydrolysis. For SDESA with ECAP, AChE already had sufficient activity in an optimized buffer; PAAS was more favorable for substrate stability and product absorbance except for lower activity. Therefore, PAAS engineered for sufficient activity plus alkaline phosphatase is absorbing for ELISA via SDESA.

  17. Comparison of SNP-based detection assays for food analysis: Coffee authentication.

    Science.gov (United States)

    Spaniolas, Stelios; Bazakos, Christos; Tucker, Gregory A; Bennett, Malcolm J

    2014-01-01

    Recently, DNA-based authentication methods were developed to serve as complementary approaches to analytical chemistry techniques. The single nucleotide polymorphism (SNP)-based reaction chemistries, when combined with the existing detection methods, could result in numerous analytical approaches, all with particular advantages and disadvantages. The dual aim of this study was (a) to develop SNP-based analytical assays such as the single-base primer extension (SNaPShot) and pyrosequencing in order to differentiate Arabica and Robusta varieties for the authentication of coffee beans and (b) to compare the performances of SNaPshot, pyrosequencing and the previously developed polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using an Agilent 2100 Bioanalyzer on the basis of linearity (R2) and LOD, expressed as percentage of the adulterant species, using green coffee beans (Arabica and Robusta) as a food model. The results showed that SNaPshot analysis exhibited the best LOD, whereas pyrosequencing revealed the best linearity (R2 = 0.997). The PCR-RFLP assay using the Agilent 2100 Bioanalyzer could prove to be a very useful method for a laboratory that lacks sequencing facilities but it can be used only if a SNP creates/deletes a restriction site.

  18. Comparison of Innovative Molecular Approaches and Standard Spore Assays for Assessment of Surface Cleanliness ▿

    Science.gov (United States)

    Cooper, Moogega; La Duc, Myron T.; Probst, Alexander; Vaishampayan, Parag; Stam, Christina; Benardini, James N.; Piceno, Yvette M.; Andersen, Gary L.; Venkateswaran, Kasthuri

    2011-01-01

    A bacterial spore assay and a molecular DNA microarray method were compared for their ability to assess relative cleanliness in the context of bacterial abundance and diversity on spacecraft surfaces. Colony counts derived from the NASA standard spore assay were extremely low for spacecraft surfaces. However, the PhyloChip generation 3 (G3) DNA microarray resolved the genetic signatures of a highly diverse suite of microorganisms in the very same sample set. Samples completely devoid of cultivable spores were shown to harbor the DNA of more than 100 distinct microbial phylotypes. Furthermore, samples with higher numbers of cultivable spores did not necessarily give rise to a greater microbial diversity upon analysis with the DNA microarray. The findings of this study clearly demonstrated that there is not a statistically significant correlation between the cultivable spore counts obtained from a sample and the degree of bacterial diversity present. Based on these results, it can be stated that validated state-of-the-art molecular techniques, such as DNA microarrays, can be utilized in parallel with classical culture-based methods to further describe the cleanliness of spacecraft surfaces. PMID:21652744

  19. Comparison of innovative molecular approaches and standard spore assays for assessment of surface cleanliness.

    Science.gov (United States)

    Cooper, Moogega; La Duc, Myron T; Probst, Alexander; Vaishampayan, Parag; Stam, Christina; Benardini, James N; Piceno, Yvette M; Andersen, Gary L; Venkateswaran, Kasthuri

    2011-08-01

    A bacterial spore assay and a molecular DNA microarray method were compared for their ability to assess relative cleanliness in the context of bacterial abundance and diversity on spacecraft surfaces. Colony counts derived from the NASA standard spore assay were extremely low for spacecraft surfaces. However, the PhyloChip generation 3 (G3) DNA microarray resolved the genetic signatures of a highly diverse suite of microorganisms in the very same sample set. Samples completely devoid of cultivable spores were shown to harbor the DNA of more than 100 distinct microbial phylotypes. Furthermore, samples with higher numbers of cultivable spores did not necessarily give rise to a greater microbial diversity upon analysis with the DNA microarray. The findings of this study clearly demonstrated that there is not a statistically significant correlation between the cultivable spore counts obtained from a sample and the degree of bacterial diversity present. Based on these results, it can be stated that validated state-of-the-art molecular techniques, such as DNA microarrays, can be utilized in parallel with classical culture-based methods to further describe the cleanliness of spacecraft surfaces.

  20. Westernization of dietary patterns among young Japanese and Polish females -- a comparison study.

    Science.gov (United States)

    Morinaka, Tomoko; Wozniewicz, Malgorzata; Jeszka, Jan; Bajerska, Joanna; Nowaczyk, Paulina; Sone, Yoshiaki

    2013-01-01

    Nowadays, the process of the westernization of eating habits is perceived to be one of the main causes of epidemics of civilization diseases, such as metabolic syndrome. The aim of the study was to assess the westernization of eating habits among 100 Japanese (aged 18-23 years) and 111 Polish female students (aged 19-25 years) of nutrition science related faculties. Food-frequency questionnaires were used to assess a dietary pattern during the four seasons of a one-year investigation. Data obtained in each season was pooled. The frequency of consumption of different foods (servings/week) between the two countries was compared and characterization of the dietary patterns of both studied populations was analyzed by factor analysis. When food consumption between the two countries was compared, apart from total meat and meat products and high-energy drink intake, significant differences were observed in all foods and food groups. Three dietary patterns were identified in both groups. Among Japanese participants, the first pattern was 'traditional Japanese', the second 'sweets and beverages', and the third 'Western', explaining 9.0%, 8.5% and 6.4% of the total variance, respectively. Among Polish participants, the first pattern was 'prudent', the second 'Western', and the third 'sweets and alcoholic beverages', explaining 8.2%, 7.7%, 6.4% of the total variance, respectively. Although the 'Western' dietary pattern was found in both groups, there were some differences in the remaining dietary patterns between the two countries. In the Japanese participants, significant cultural influences on habitual food intake could still be observed, and the extent of diet westernization seems to be smaller compared to the Polish participants.

  1. Comparison of Chinese medicine higher education programs in China and five Western countries

    Directory of Open Access Journals (Sweden)

    Pei Xue

    2015-10-01

    Conclusions: There are large differences among their curriculum and enrollment policies. This comparison should provide information about the further development of international standards in TCM education.

  2. Comparison of two enzyme-linked immunosorbent assays and one rapid immunoblot assay for detection of herpes simplex virus type 2-specific antibodies in serum

    NARCIS (Netherlands)

    Groen, J; Van Dijk, G; Niesters, H G; Van Der Meijden, W I; Osterhaus, A D

    The sensitivities and specificities of three immunoassays for the detection of herpes simplex virus type 2 (HSV-2)-specific immunoglobulin G antibodies in serum, including the one-strip rapid immunoblot assay (RIBA; Chiron Corporation) and two indirect enzyme immunosorbent assays (EIA; Gull

  3. A Comparison of Anti-Nuclear Antibody Quantification Using Automated Enzyme Immunoassays and Immunofluorescence Assays

    DEFF Research Database (Denmark)

    Baronaite, Renata; Engelhart, Merete; Mørk Hansen, Troels;

    2014-01-01

    Anti-nuclear antibodies (ANA) have traditionally been evaluated using indirect fluorescence assays (IFA) with HEp-2 cells. Quantitative immunoassays (EIA) have replaced the use of HEp-2 cells in some laboratories. Here, we evaluated ANA in 400 consecutive and unselected routinely referred patients...... patients were compared. The majority of the results were the same between the two methods (n = 325, 84%); however, 8% (n = 30) yielded equivocal results (equivocal-negative and equivocal-positive) and 8% (n = 31) yielded divergent results (positive-negative). The results showed fairly good agreement......, with Cohen's kappa value of 0.30 (95% confidence interval (CI) = 0.14-0.46), which decreased to 0.23 (95% CI = 0.06-0.40) when the results for dsDNA were omitted. The EIA method was less reliable for assessing nuclear and speckled reactivity patterns, whereas the IFA method presented difficulties detecting...

  4. Comparison of DNA Fragmentation Assay in Frozen-Thawed Cat Epididymal Sperm.

    Science.gov (United States)

    Kunkitti, P; Sjödahl, A; Bergqvist, A-S; Johannisson, A; Axnér, E

    2016-08-01

    DNA fragmentation of frozen-thawed feline epididymal sperm from corpus and cauda regions was evaluated by three different techniques. The DNA fragmentation index (DFI) was compared between techniques: the sperm chromatin structural assay (SCSA(®) ), acridine orange staining techniques (AOT) and the sperm chromatin dispersion (SCD). There were significant differences in DFI among the techniques (p < 0.05) with no correlations. Only DFI values obtained from SCD revealed a significantly higher DFI in corpus compared with cauda spermatozoa (p < 0.05). The discrepancy between techniques might be due to the sensitivity of each technique, differences in severity of DNA damaged that can be detected. The difference in DFI between epididymal regions from SCD technique might indicate different maturational stages of spermatozoa, with less chromatin condensation of spermatozoa in corpus compared with cauda epididymis. © 2016 Blackwell Verlag GmbH.

  5. Comparison of in vitro hormone activities of selected phthalates using reporter gene assays.

    Science.gov (United States)

    Shen, Ouxi; Du, Guizhen; Sun, Hong; Wu, Wei; Jiang, Yi; Song, Ling; Wang, Xinru

    2009-12-01

    Phthalates are widely used in the plastic industry and food packaging, imparting softness and flexibility to normally rigid plastic medical devices and children's toys. Even though phthalates display low general toxicity, there is increasing concern on the effects of endocrine system induced by some of phthalate compounds. The hormone activity of dibutyl phthalate (DBP), mono-n-butyl phthalate (MBP) and di-2-ethylhexyl phthalate (DEHP) were assessed using the luciferase reporter gene assays. The results showed that DBP, MBP and DEHP, not only exhibited potent antiandrogenic activity, with IC(50) value of 1.05x10(-6), 1.22x10(-7)M and exceeding 1x10(-4)M respectively, but also showed the androgenic activity with EC(50) value of 6.17x10(-6), 1.13x10(-5)M and exceeding 1x10(-4)M. We also found that all the three related chemicals possessed thyroid receptor (TR) antagonist activity with IC(50) of 1.31x10(-5), 2.77x10(-6)M and exceeding 1x10(-4)M respectively, and none showed TR agonist activity. These results indicate that TR might be the targets of industrial chemicals. In the ER mediate reporter gene assay, three chemicals showed no agonistic activity except for DBP, which appeared weakly estrogenic at the concentration of 1.0x10(-4)M. Together, the findings demonstrate that the three phthalates could simultaneously disrupt the function of two or more hormonal receptors. Therefore, these phthalates should be considered in risk assessments for human health.

  6. Framing the Iraq war: a cross-national comparison of newspaper framing in four Western countries

    NARCIS (Netherlands)

    Vliegenthart, R.; Schröder, H.

    2010-01-01

    In this paper we compare the newspaper attention for and framing of the Iraq issue in four Western countries (US, UK, Germany and the Netherlands) during the period September 2002 until August 2003. Using computer assisted coding based on wordlists constructed by human coders, we analyzed more than

  7. The Persian Reception of Western Law and its Subsequent Organic Development in Inner-Asian Comparison

    DEFF Research Database (Denmark)

    Afsah, Ebrahim

    This presentation thus seeks to investigate the active engagement of a non-Western society during 19th and early 20th century with a legal tradition they identified as simultaneously more powerful and sharply at odds with its own. It seeks to develop commonalities and differences in the colonial...

  8. Comparison of GMT presto assay and Roche cobas® 4800 CT/NG assay for detection of Chlamydia trachomatis and Neisseria gonorrhoeae in dry swabs.

    Science.gov (United States)

    de Waaij, Dewi J; Dubbink, Jan Henk; Peters, Remco P H; Ouburg, Sander; Morré, Servaas A

    2015-11-01

    Urogenital Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) are the most prevalent bacterial STIs worldwide. Molecular tests are the standard for the detection of CT and NG, as these are difficult to culture. The recently introduced CE-IVD marked GMT Presto assay promises to be a valuable addition in CT and NG diagnostics. The advantage of the Presto assay is that it works on many PCR systems and the DNA can be isolated by any system.We compared the Presto assay to the widely used Roche cobas® 4800 CT/NG test for the detection of CT and NG in 612 vaginal and rectal dry collected swabs. Discrepant samples were tested by the TIB MOLBIOL Lightmix Kit 480 HT CT/NG assay. The alloyed gold standard was defined as two concurring Presto and cobas® 4800 results, or, with discrepant Presto and cobas® results, two concurring results of either test together with the Lightmix Kit 480 HT CT/NG assay. For the Presto assay,we observed 77 CT positive (13%) and 22 NG positive (3,6%) vaginal samples, and 41 CT positive (6,7%) and 11 NG positive (1,8%) rectal samples. For the cobas® 4800 assay,we observed 77 CT positive (13%) and 21NG positive (3,4%) vaginal samples, and 39 CT positive (6,4%) and 11 NG positive (1,8%) rectal samples. Ten CT samples were discrepant between Presto and cobas® 4800 CT/NG assays, while two NG samples were discrepant. CT sensitivity in both assays was 100% compared to the alloyed gold standard. The sensitivity was 100% for both vaginal and rectal dry swabs, underlining the suitability of these sample types for detection of CT and NG. The Presto assay is therefore valuable for molecular detection of CT and NG in dry vaginal and rectal swabs.

  9. Comparison between indigenous and Western postnatal care practices in Mopani District, Limpopo Province, South Africa

    Directory of Open Access Journals (Sweden)

    Roinah N. Ngunyulu

    2015-02-01

    Full Text Available Background: Postnatal care begins immediately after the expulsion of the placenta and continues for six to eight weeks post-delivery. High standard of care is required during the postnatal period because mothers and babies are at risk and vulnerable to complications related to postpartum haemorrhage and infections. Midwives and traditional birth attendants are responsible for the provision of postnatal care in different settings, such as clinics and hospitals, and homes.Methods: A qualitative, exploratory, descriptive and contextual research approach was followed in this study. Unstructured interviews were conducted with the traditional birth attendants. An integrated literature review was conducted to identify the Western postnatalcare practices. Tesch’s process was followed during data analysis.Findings: The following main categories were identified: similarities between indigenous and Western postnatal care practices, and differences between indigenous and Western postnatal care practices. Based on these findings, training of midwives and traditional birth attendants was recommended in order to empower them with knowledge and skills regarding the indigenous and Western postnatal care practices.Conclusions: It is evident that some indigenous postnatal care practices have adverse effects on the health of postnatal women and their newborn infants, but these are unknown to the traditional birth attendants. The employment of indigenous postnatal care practices by the traditional birth attendants is also influenced by their cultural beliefs, norms, values and attitudes. Therefore, there is an urgent need to train midwives and traditional birth attendants regarding the indigenous and Western postnatal care to improve the health of postnatal women and their babies.

  10. 中西方酒文化比较%A Comparison between Chinese and Western Wine Cul-ture

    Institute of Scientific and Technical Information of China (English)

    何凤玲

    2014-01-01

    As a beverage, the wine is infiltrated into almost all human activities and plays an important role. It is not only an objective material, but also a symbol of a kind of culture. In to-day's frequent Chinese and Western cultural communication, the differences of Chinese and Western wine culture have been attached more and more importance. Through a comparison of the origin of wine, the morality and etiquette of wine, and wine-making raw materials between China and West, the dif-ferences between Chinese and Western wine culture are ana-lyzed, in order to avoid embarrassments in cross-cultural com-munication.%作为一种饮品,酒几乎渗透在人类一切活动中,并在其中起着重要作用。它不仅是一种客观物质,且代表一种文化。在中西方文化频繁交流的今天,中西酒文化之间的差异越来越受到重视。文章通过对中西方酒的起源、酒德酒礼和酿酒原料的对比,来探讨中西方酒文化差异,以避免在跨文化交际中陷入尴尬。

  11. Comparison of integrated Chinese and Western medicine with and without somatostatin supplement in the treatment of severe acute pancreatitis

    Institute of Scientific and Technical Information of China (English)

    Qing Xia; Lin Yuan; Xiao-Nan Yang; Wen-Fu Tang; Jun-Ming Jiang

    2005-01-01

    AIM: To evaluate the therapeutic effect of the combined use of early short-term somatostatin and conventional integrated Chinese and Western medicine in treating severe acute pancreatitis.METHODS: Sixty patients with severe acute pancreatitis were divided at random into a somatostatin group and a basic treatment group. Both groups received integrated traditional Chinese and Western medicine without surgery.For patients in the somatostatin group, somatostatin was infused intravenously 250 μg/h for 72 h; other medications were the same as in the basic treatment group. In both groups, comparisons of therapeutic effectiveness were made in terms of morbidity of organic dysfunction and mortality rate, and severity of the disease according to serum levels of C-reaction protein, scores of acute physiology and chronic health evaluation (APACHE Ⅱ), and scores of Balthazar-CT.RESULTS: The indexes for C-reaction protein levels on the fourth and seventh days, and APACHE Ⅱ scores on the seventh day after treatment, were significantly improved in the somatostatin group than in the basic treatment group. The morbidity of organic dysfunction was lower in the somatostatin group than in the basic treatment group, although the difference was not statistically significant. There was no significant difference in mortality between the two groups.CONCLUSION: We conclude that combined traditional Chinese and Western medicines with an early short-term use of somatostatin can improve the condition of patients with severe acute pancreatitis.

  12. Comparison of Parasite Burden Using Real-Time Polymerase Chain Reaction Assay and Limiting Dilution Assay in Leishma-nia major Infected Mouse

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    Somayeh GHOTLOO

    2015-12-01

    Full Text Available Background:Limiting dilution assay is considered as the gold standard method for quantifying the number of parasites in the animal model of Leishmania infection. Nowadays, real-time PCR is being increasingly applied to quantify infectious agents. In the present study, a real-time PCR assay was developed to estimate para­site burdens in lymph nodes of Leishmania major infected BALB/C mice. Enumera­tion of parasites was also performed by limiting dilution assay and compared with the results of real-time PCR based quantification.Methods:The SYBR Green based real- time PCR assay was performed to amplify a 75 bp fragment of superoxide dismutase B1 gene in the lymph nodes of L. major infected BALB/C mice 8 weeks post infection. Mice were infected subcutaneously at the base of their tail with 2 × 105L. major promastigotes in the stationary phase of growth. To compare parasite burdens obtained by real-time PCR assay with those of limiting dilution assay, twelve 8-fold serial dilutions of the lymph node homoge­nates were prepared in the Schneider medium and incubated at 26°C.After 7 days, wells containing motile parasites were identified by direct observation under an inverted light microscope and the total number of parasites was estimated using the ELIDA software.Results:Spearman's correlation coefficient of the parasite burdens between real-time PCR and limiting dilution assay was 0.72 (Pvalue = 0.008.Conclusion:Real-time PCR assay is an appropriate replacement to existing limit­ing dilution assay in quantifying parasite burden in the experimental model of Leishma­nia infection.

  13. Comparison of in vitro cell culture and a mouse assay for measuring infectivity of Cryptosporidium parvum.

    Science.gov (United States)

    Rochelle, Paul A; Marshall, Marilyn M; Mead, Jan R; Johnson, Anne M; Korich, Dick G; Rosen, Jeffrey S; De Leon, Ricardo

    2002-08-01

    In vitro cell cultures were compared to neonatal mice for measuring the infectivity of five genotype 2 isolates of Cryptosporidium parvum. Oocyst doses were enumerated by flow cytometry and delivered to animals and cell monolayers by using standardized procedures. Each dose of oocysts was inoculated into up to nine replicates of 9 to 12 mice or 6 to 10 cell culture wells. Infections were detected by hematoxylin and eosin staining in CD-1 mice, by reverse transcriptase PCR in HCT-8 and Caco-2 cells, and by immunofluorescence microscopy in Madin-Darby canine kidney (MDCK) cells. Infectivity was expressed as a logistic transformation of the proportion of animals or cell culture wells that developed infection at each dose. In most instances, the slopes of the dose-response curves were not significantly different when we compared the infectivity models for each isolate. The 50% infective doses for the different isolates varied depending on the method of calculation but were in the range from 16 to 347 oocysts for CD-1 mice and in the ranges from 27 to 106, 31 to 629, and 13 to 18 oocysts for HCT-8, Caco-2, and MDCK cells, respectively. The average standard deviations for the percentages of infectivity for all replicates of all isolates were 13.9, 11.5, 13.2, and 10.7% for CD-1 mice, HCT-8 cells, Caco-2 cells, and MDCK cells, respectively, demonstrating that the levels of variability were similar in all assays. There was a good correlation between the average infectivity for HCT-8 cells and the results for CD-1 mice across all isolates for untreated oocysts (r = 0.85, n = 25) and for oocysts exposed to ozone and UV light (r = 0.89, n = 29). This study demonstrated that in vitro cell culture was equivalent to the "gold standard," mouse infectivity, for measuring the infectivity of C. parvum and should therefore be considered a practical and accurate alternative for assessing oocyst infectivity and inactivation. However, the high levels of variability displayed by all

  14. Comparison of three typing assays for nicotinamide adenine dinucleotide-independent Actinobacillus pleuropneumoniae.

    Science.gov (United States)

    Maldonado, Jaime; Blanco, Mónica; Martínez, Eva; Navas, Jesús

    2011-07-01

    Three tests for typing clinical isolates of Actinobacillus pleuropneumoniae biovar 2 were compared: 1) standard coagglutination with type-specific antisera against serovars 1-12 of biovar 1 of A. pleuropneumoniae; 2) a previously described polymerase chain reaction system for detecting the apx genes encoding the ApxI, ApxII, and ApxIII toxins in A. pleuropneumoniae; and 3) a restriction fragment length polymorphism analysis of the transferrin-binding protein B gene. The panel of strains tested included 112 field isolates of biovar 2 recovered from pigs between 1979 and 2007 in Italy and Spain, and reference strains for all described serovars of both biovars. The values of Simpson index of diversity obtained for the 3 methods were 0.68, 0.20, and 0.60, respectively. Coagglutination assays identified the field isolates as belonging to serovars 2 (9 strains), 4 (13 strains), 7 (61 strains), 9 (17 strains), and 11 (1 strain). Eleven strains were not typeable, and cross-reactivity was observed between serovars 2 and 4, 4 and 7, and 9 and 11. Isolates of A. pleuropneumoniae biovar 2 displayed 2 apx patterns: ApxII(+) (94 strains) and ApxI(+)/ApxII(+) (18 strains). The restriction fragment length polymorphism analysis assigned the strains tested to 3 different patterns. This method distinguished between biovar 2 reference strains and field strains that could not be identified by other methods, thus constituting a useful complementary test for the typing of A. pleuropneumoniae biovar 2.

  15. Comparison of an ELISA assay for the detection of adhesive/invasive Neospora caninum tachyzoites.

    Science.gov (United States)

    Pereira, Luiz Miguel; Yatsuda, Ana Patrícia

    2014-03-01

    Neospora caninum belongs to the phylum Apicomplexa, the causative agent of neosporosis, which leads to economic impacts on cattle production. A common feature among apicomplexan parasites is the invasive process driven mostly by the parasite. As a first evaluation of candidate molecules that play a possible role by interfering in this invasive process, the in vitro invasion assay is a fast and direct way to screen future agonists or antagonists. This work involved the development of a new cell culture ELISA and transient β-galactosidase activity applied to the semi-quantitative detection of N. caninum in Vero cell culture. Cell culture ELISA is based on histochemistry and immunology, resulting in a colorimetric reaction. The β-galactosidase activity was obtained by the transient transfection of the lacZ gene under control of RPS13 promoter of N. caninum. These methods were used to evaluate the effects of temperature (37°C and 85°C) on the invasion and adhesion of tachyzoites. The three tested methods (real time PCR, β-galactosidase activity and ELISA) showed a similar pattern, indicating that different methods may be complementary.

  16. Comparison of frozen versus desiccated reference human red blood cells for hemagglutination assays.

    Science.gov (United States)

    Ho, David; Schierts, Jennifer; Zimmerman, Zea; Gadsden, Isaac; Bruttig, Stephen

    2009-10-01

    Red blood cells (RBCs) are commonly used fresh or stored in frozen format for identification of patients' antibodies and serologic specificity of such antibodies at reference laboratories. However, maintaining a large pool of fresh RBCs is impossible in a blood-banking environment and blood in frozen format poses a logistic disadvantage in terms of accessibility, maintenance cost, safety, and sample recovery. This study explores an alternative, desiccation storage method for RBCs to provide a reagent that supports greater utilization and flexibility for reference laboratories. RBCs from five donors were used in the study. RBCs were processed and kept in either frozen or desiccated format. Study variables for either the frozen or the desiccated cells included cell recovery as quantified by cell counts, gross microscopic examination, and hemagglutination assays. The mean percentage of cell recovery for thawed and washed frozen RBCs was 20% versus 50% for rehydrated and washed desiccated RBCs. Microscopic examination of thawed cells from the frozen preparation showed cells with irregular shapes, a sharp contrast when compared with rehydrated cells from the desiccated preparation, where cells are mostly intact, smooth surface, and biconcave in structure. Cells in both preparations performed well in manual agglutination tests. Desiccation preservation of RBCs provides a somewhat better RBC recovery and cell structure stability, while maintaining the necessary antigen-antibody reactions for cell surface markers, which will allow desiccated RBCs to be archived in blood collecting and processing reference laboratories.

  17. Comparison of an ELISA assay for the detection of adhesive/invasive Neospora caninum tachyzoites

    Directory of Open Access Journals (Sweden)

    Luiz Miguel Pereira

    Full Text Available Neospora caninum belongs to the phylum Apicomplexa, the causative agent of neosporosis, which leads to economic impacts on cattle production. A common feature among apicomplexan parasites is the invasive process driven mostly by the parasite. As a first evaluation of candidate molecules that play a possible role by interfering in this invasive process, the in vitro invasion assay is a fast and direct way to screen future agonists or antagonists. This work involved the development of a new cell culture ELISA and transient β-galactosidase activity applied to the semi-quantitative detection of N. caninum in Vero cell culture. Cell culture ELISA is based on histochemistry and immunology, resulting in a colorimetric reaction. The β-galactosidase activity was obtained by the transient transfection of the lacZ gene under control of RPS13 promoter of N. caninum. These methods were used to evaluate the effects of temperature (37°C and 85°C on the invasion and adhesion of tachyzoites. The three tested methods (real time PCR, β-galactosidase activity and ELISA showed a similar pattern, indicating that different methods may be complementary.

  18. Diagnostic tests for amoebic liver abscess: comparison of enzyme - linked immunosorbent assay (Elisa and counterimmunoelectrophoresis (CIE

    Directory of Open Access Journals (Sweden)

    Marcos I. Restrepo

    1996-02-01

    Full Text Available The liver abscess is the most frequent extraintestinal complication of intestinal amoebiasis: its diagnosis is suggested by the clinical picture but it must be confirmed by paraclinic tests. Themost stringent diagnosis requires identification of E. histolytica. But this is possible only in a few cases. Serological tests greatly improve the diagnosis of this severe complication of amoebiasis. We compared the Enzyme Linfed Immunosorbent Assay and the Counterimmunoeletrophoresis techniques. Both techniques were used to detect amoebic antibodies in 50 control patients, 30 patients with liver abscess and 30 patients with intestinal amoebiasis. All the sera from control patients gave negative results iin both techniques. When analysing the sera from patients with intestinal amoebiasis, 10% of them were positive by ELISA but non by CIE. The sera of patients with liver abscess, we found that 90% were positive by the ELISA method and 66.6% by the CIE technique. In patients with amoebic liver abscess, the results showed that the ELISA was more sensitive than the CIE, as it presented a higher sensitivity (100% than that of the CIE technique (66%.

  19. Are extraction methods in quantitative assays of pharmacopoeia monographs exhaustive? A comparison with pressurized liquid extraction.

    Science.gov (United States)

    Basalo, Carlos; Mohn, Tobias; Hamburger, Matthias

    2006-10-01

    The extraction methods in selected monographs of the European and the Swiss Pharmacopoeia were compared to pressurized liquid extraction (PLE) with respect to the yield of constituents to be dosed in the quantitative assay for the respective herbal drugs. The study included five drugs, Belladonnae folium, Colae semen, Boldo folium, Tanaceti herba and Agni casti fructus. They were selected to cover different classes of compounds to be analyzed and different extraction methods to be used according to the monographs. Extraction protocols for PLE were optimized by varying the solvents and number of extraction cycles. In PLE, yields > 97 % of extractable analytes were typically achieved with two extraction cycles. For alkaloid-containing drugs, the addition of ammonia prior to extraction significantly increased the yield and reduced the number of extraction cycles required for exhaustive extraction. PLE was in all cases superior to the extraction protocol of the pharmacopoeia monographs (taken as 100 %), with differences ranging from 108 % in case of parthenolide in Tanaceti herba to 343 % in case of alkaloids in Boldo folium.

  20. A fully validated microbiological assay for daptomycin injection and comparison to HPLC method

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    Ana Paula Christ

    2015-12-01

    Full Text Available abstract Daptomycin (DPT was the first lipopeptide antibiotic available for commercialization. It is active against gram-positive bacteria, including resistant strains. This work aimed to develop and validate a turbidimetric microbiologic assay to determine daptomycin in an injectable form. A 3x3 design was employed, at concentrations of 1, 2 and 4.0 µg/mL. The microorganism test used was Staphylococcus aureus ATCC 6538p, and Antibiotic Medium 3 was used as the culture medium. Method validation demonstrated that the bioassay was linear (r=0.9995, precise (RSD=2.58%, accurate (recovery 100.48± 2.11%, and robust. Degradation kinetics was also performed in an alkaline medium, indicating that daptomycin degradation follows first order kinetics under these conditions. The analyses of degraded solutions showed that daptomycin degradation products do not possess bactericidal activity. The bioassay was compared to HPLC method that was previously developed and no significant difference was found between them (p>0.05. The method proved to be appropriate for daptomycin injection quality control.

  1. Comparison of molecular assays for detection and typing of human papillomavirus.

    Science.gov (United States)

    Koidl, Christoph; Bozic, Michael; Hadzisejdic, Ita; Grahovac, Maja; Grahovac, Blazenka; Kranewitter, Wolfgang; Marth, Egon; Kessler, Harald H

    2008-08-01

    The objective of the study was to compare the performance of 3 different extraction instruments in conjunction with 4 different amplification and detection kits for detection and typing of human papillomavirus (HPV) deoxyribonucleic acid (DNA). A total of 42 cervical swabs were investigated. HPV DNA was extracted on the 3 different instruments. Each of the extracts was then amplified, and HPV DNA amplification products were detected with 4 different kits. In 31 samples, HPV DNA was detected by both the Amplicor HPV test and the LINEAR ARRAY HPV genotyping test in conjunction with DNA extraction on the easyMAG instrument. In another 6 samples, only low-risk types were detected with the linear array HPV genotyping test. After extraction on the easyMAG instrument, 32 samples tested positive when the PapilloCheck with the HotStarTaq DNA polymerase was used. Together with extraction on the easyMAG instrument, the Amplicor HPV test, the linear array HPV genotyping test, and the new PapilloCheck with the HotStarTaq DNA polymerase provide comparable results allowing reliable and safe HPV diagnostics in the routine laboratory. Use of alternative assays may lead to an increase of invalid and divergent HPV typing results.

  2. Platinum-group elements in the Eastern Deccan volcanic province and a comparison with platinum metals of the western Deccan

    Indian Academy of Sciences (India)

    James Crocket; Dalim Paul; Trisha Lala

    2013-08-01

    This study is the first detailed investigation of the platinum-group elements (PGE) at the eastern margin of the Deccan volcanic province of India. One of the PGE, osmium, is not included largely because of analytical problems. The study is focused on mafic volcanics and dykes from four areas including Amarkantak, Umaria, Shahdol and Chirimiri. The first two localities represent two lava piles of about 170 and 400 m thickness respectively. In Umaria, 16 flows have been demarcated based on petrography and field studies. The Shahdol samples are basal lava formations overlying Gondwana sediments (Carboniferous) and the Chirimiri samples are dykes. In this study, the western Deccan province is defined as the Western Ghats plus Kutch. On average, the PGE are ∼20% higher in Amarkantak than Umaria and the flows are ∼13% higher in PGE than the dykes. A Zr vs. Pd scattergram found a strong positive correlation for these two elements except for one Umaria sample which indicated severe Pd loss. A comparison of west and east parts of the Deccan volcanic province using primitive mantle normalization showed that higher values prevailed in the western province suite in the Ni-Ir-Ru-Pt region. In contrast, eastern province values dominated in the Pd-Au-Cu region at the ‘Cu’ end of the profiles. A strong dominance of Pd in the eastern Deccan was also of interest. A number of factors, for example, percentage partial melting of the source rock and the temperature and pressure of partial melting strongly influence the character of these profiles. The observed PGE profile characteristics probably result in part from a long distance of subsurface transport of Deccan magma from the western to eastern regions.

  3. Exotic annual Bromus invasions: comparisons among species and ecoregions in the western United States

    Science.gov (United States)

    Brooks, Matthew L.; Brown, Cynthia S.; Chambers, Jeanne C.; D'Antonio, Carla M.; Keeley, Jon E.; Belnap, Jayne

    2016-01-01

    Exotic annual Bromus species are widely recognized for their potential to invade, dominate, and alter the structure and function of ecosystems. In this chapter, we summarize the invasion potential, ecosystem threats, and management strategies for different Bromus species within each of five ecoregions of the western United States. We characterize invasion potential and threats in terms of ecosystem resistance to Bromus invasion and ecosystem resilience to disturbance with an emphasis on the importance of fi re regimes. We also explain how soil temperature and moisture regimes can be linked to patterns of resistance and resilience and provide a conceptual framework that can be used to evaluate the relative potential for invasion and ecological impact of the dominant exotic annual Bromus species in the western United States.

  4. Comparison of kDNA PCR-hybridization assay with three PCR methods for canines visceral Leishmaniasis diagnosis

    Energy Technology Data Exchange (ETDEWEB)

    Pilatti, Marcia M.; Andrade, Antero S.R. [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN-MG), Belo Horizonte, MG (Brazil)], e-mail: marciapilatti@yahoo.com.br, e-mail: antero@cdtn.br; Ferreira, Sidney A. [Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, MG (Brazil). Dept. de Parasitologia], e-mail: saninoalmeida@gmail.com

    2009-07-01

    The sensitivity of the kDNA PCR-Hybridization assay, which uses radioactive DNA probes (labeled with {sup 32}P), was compared with three conventional PCR methods used for canine visceral leishmaniasis diagnosis. All PCR methods had two steps: a first amplification followed by hybridization or by a new amplification (nested or semi nested). Two methods (kDNA PCR-Hybridization and kDNA snPCR) used primers addressed to kinetoplast minicircles and the other two methods to the coding (LnPCR) and intergenic noncoding regions (ITS-1 nPCR) of the ribosomal rRNA genes. The comparison was accomplished in two groups of 23 infected dogs using samples collected by the conjunctival swab procedure. In the Group 1 the DNA was extracted from cotton swabs by phenol-chloroform and in Group 2 by boiling. The most efficient PCR methods in the Group 1 were those based on kDNA targets. The kDNA PCR-Hybridization was able to detect parasites in 22/23 dogs (95.6%) and in 40/46 samples (86.9%). The kDNA snPCR was positive for 21/23 dogs (91.3%) and for 40/46 samples (86.9%). The positivities of the kDNA based methods were significantly higher than the positivities verified for the methods based on ribosomal rRNA genes (p<0.05). In the Group 2 the kDNA PCR- Hybridization showed a better performance detecting parasites in 18/23 dogs (78.3%) and in 31/46 samples (67.4%), significantly higher than the other three methods (p<0.05). The higher sensitivity of the minicircle kDNA based assays reported by others was confirmed in this study and kDNA PCR-Hybridization showed the best sensitivity among the assays evaluated. (author)

  5. Comparison of T7E1 and surveyor mismatch cleavage assays to detect mutations triggered by engineered nucleases.

    Science.gov (United States)

    Vouillot, Léna; Thélie, Aurore; Pollet, Nicolas

    2015-01-07

    Genome editing using engineered nucleases is used for targeted mutagenesis. But because genome editing does not target all loci with similar efficiencies, the mutation hit-rate at a given locus needs to be evaluated. The analysis of mutants obtained using engineered nucleases requires specific methods for mutation detection, and the enzyme mismatch cleavage method is used commonly for this purpose. This method uses enzymes that cleave heteroduplex DNA at mismatches and extrahelical loops formed by single or multiple nucleotides. Bacteriophage resolvases and single-stranded nucleases are used commonly in the assay but have not been compared side-by-side on mutations obtained by engineered nucleases. We present the first comparison of the sensitivity of T7E1 and Surveyor EMC assays on deletions and point mutations obtained by zinc finger nuclease targeting in frog embryos. We report the mutation detection limits and efficiencies of T7E1 and Surveyor. In addition, we find that T7E1 outperforms the Surveyor nuclease in terms of sensitivity with deletion substrates, whereas Surveyor is better for detecting single nucleotide changes. We conclude that T7E1 is the preferred enzyme to scan mutations triggered by engineered nucleases.

  6. Clinical characteristics of patients with serrated polyposis syndrome in Korea: comparison with Western patients.

    Science.gov (United States)

    Kim, Eun Ran; Jeon, Jaryong; Lee, Jin Hee; Lee, Yoon Jung; Hong, Sung Noh; Chang, Dong Kyung; Kim, Young-Ho

    2017-07-01

    Serrated polyposis syndrome (SPS) has been shown to increase the risk of colorectal cancer (CRC). However, little is known about the characteristics of Asian patients with SPS. This study aimed to identify the clinicopathological features and risk of CRC in Korean patients with SPS as well as the differences between Korean and Western patients based on a literature review. This retrospective study included 30 patients with SPS as defined by World Health Organization classification treated at Samsung Medical Center, Korea, between March 1999 and May 2011. Twenty patients (67%) were male. The median patient age at diagnosis was 56 years (range, 39-76 years). A total of 702 polyps were identified during a median follow-up of 43 months (range, 0-149 months). Serrated polyps were noted more frequently in the distal colon (298/702, 55%). However, large serrated polyps and serrated adenomas were mainly distributed throughout the proximal colon (75% vs. 25% and 81% vs. 19%, respectively); 73.3% had synchronous adenomatous polyps. The incidence of CRC was 10% (3/30 patients), but no interval CRC was detected. A total of 87% of the patients underwent esophagogastroduodenoscopy and 19.2% had significant lesions. The phenotype of SPS in Korean patients is different from that of Western patients. In Korean patients, SPS is more common in men, there were fewer total numbers of serrated adenoma/polyps, and the incidence of CRC was lower than that in Western patients. Korean patients tend to more frequently have abnormal gastric lesions. However, the prevalence of synchronous adenomatous polyps is high in both Western and Korean patients.

  7. Oral microbiome profiles: 16S rRNA pyrosequencing and microarray assay comparison.

    Directory of Open Access Journals (Sweden)

    Jiyoung Ahn

    Full Text Available OBJECTIVES: The human oral microbiome is potentially related to diverse health conditions and high-throughput technology provides the possibility of surveying microbial community structure at high resolution. We compared two oral microbiome survey methods: broad-based microbiome identification by 16S rRNA gene sequencing and targeted characterization of microbes by custom DNA microarray. METHODS: Oral wash samples were collected from 20 individuals at Memorial Sloan-Kettering Cancer Center. 16S rRNA gene survey was performed by 454 pyrosequencing of the V3-V5 region (450 bp. Targeted identification by DNA microarray was carried out with the Human Oral Microbe Identification Microarray (HOMIM. Correlations and relative abundance were compared at phylum and genus level, between 16S rRNA sequence read ratio and HOMIM hybridization intensity. RESULTS: The major phyla, Firmicutes, Proteobacteria, Bacteroidetes, Actinobacteria, and Fusobacteria were identified with high correlation by the two methods (r = 0.70∼0.86. 16S rRNA gene pyrosequencing identified 77 genera and HOMIM identified 49, with 37 genera detected by both methods; more than 98% of classified bacteria were assigned in these 37 genera. Concordance by the two assays (presence/absence and correlations were high for common genera (Streptococcus, Veillonella, Leptotrichia, Prevotella, and Haemophilus; Correlation = 0.70-0.84. CONCLUSION: Microbiome community profiles assessed by 16S rRNA pyrosequencing and HOMIM were highly correlated at the phylum level and, when comparing the more commonly detected taxa, also at the genus level. Both methods are currently suitable for high-throughput epidemiologic investigations relating identified and more common oral microbial taxa to disease risk; yet, pyrosequencing may provide a broader spectrum of taxa identification, a distinct sequence-read record, and greater detection sensitivity.

  8. Comparison of two methods (microscopy and enzyme-linked immunosorbent assay) for the diagnosis of amebiasis.

    Science.gov (United States)

    Tanyuksel, Mehmet; Yilmaz, Hasan; Ulukanligil, Mustafa; Araz, Engin; Cicek, Mutalip; Koru, Ozgur; Tas, Zeynep; Petri, William A

    2005-07-01

    Diagnosis of amebiasis is usually performed on a clinical basis alone in most endemic countries having limited economic resources. This epidemiological study was conducted using modern diagnostic tests for amebiasis in the southeastern region of Turkey, an endemic area for amebiasis. The population of this study included patients with symptomatic diarrhea/dysentery attending both Yuzuncu Yil University, Van and Harran University, Sanliurfa, Turkey. A total of 380 stool specimens were collected and examined for Entamoeba by light microscopy (fresh, lugol, and trichrome staining) and stool antigen detection based- enzyme-linked immunosorbent assay (EIA) test (TechLab Entamoeba histolytica II). 24% (91/380) of stool specimens were positive for E. histolytica/Entamoeba dispar trophozoites/cysts microscopically using trichrome staining. 13% (51/380) of the stool specimens were found to be positive for E. histolytica by the EIA test, including 15% (14/91) of microscopy (+) stool specimens and 13% (37/289) of microscopy (-) stool specimens. Enteric parasites were common in these populations with 66% (251/380) of the study population harboring more than one parasite. In addition to the 13% (51/380) of patients determined to have E. histolytica by EIA, eighty-six patients (22.6%) had Blastocystis hominis, 54 (14.2%) Entamoeba coli, 44 (11.5%) Giardia lamblia, 16 (4.2%) Chilomastix mesnili, 15 (3.9%) Iodamoeba bütschlii, 12 (3.1%) Hymenolepis nana, 9 (2.3%) Endolimax nana, 9 (2.3%) Dientamoeba fragilis, and 8 (2.1%) had Ascaris lumbricoides. We concluded that E. histolytica infection was found in 13% of the patients presenting with diarrhea in Van and Sanliurfa Turkey.

  9. Comparison of fumonisin contamination using HPLC and ELISA methods in Bt and near-isogenic maize hybrids infested with European corn borer or Western bean cutworm

    Science.gov (United States)

    Field trials were conducted (2007 to 2010) to compare grain fumonisin levels among non-Bt maize hybrids and Bt hybrids with transgenic protection against European corn borer and Western bean cutworm (WBC). High-performance liquid chromatography (HPLC) and enzyme-linked immunosorbent assay (ELISA) w...

  10. Comparison of enzyme linked immunosorbent assay (ELISA with indirect immunofluorescence for detection of anti-nuclear antibody

    Directory of Open Access Journals (Sweden)

    G.L.S. Sumanth Kumar

    2014-10-01

    Full Text Available Background: Detection of antinuclear antibody (ANA is used as one of the diagnostic criteria for autoimmune rheumatic diseases (ARD. Both indirect immunofluorescence (IIF and enzyme linked immunosorbant assay (ELISA methods are used for this purpose. However, there are lack of data comparing these two tests from India. Methods: We prospectively studed 294 patients clinically suspected to be having ARD between April 2012 and September 2013. They were tested for ANA by IIF and ELISA methods. Representative samples positive by both the tests were processed again by a line immunoassay test to detect the specific antinuclear antibodies. Considering the IIF results as the ‘gold standard’, the utility of ELISA for ANA detection was analyzed. Results: Of the 294 samples processed, 181 (61.5% were from female patients. By IIF 30% of samples in males and 40.3% sample in females tested positive. We found ELISA to have a poor sensitivity (45.8% but good specificity (99.5%. The positive predictive value for ELISA were 98% and negative predictive value 76.2% respectively. Forty four samples positive by both IIF and ELISA were tested by Western blot to detect individual autoantibodies. Of these, only 24 samples showed the presence of one or more bands, while the remaining 20 (45.4% were negative by line immunoassay. In our study anti-nuclear ribonucleoprotein/Smith was the most common ANA detected. Conclusions: The poor sensitivity raises concerns regarding the practice of initial screening for ANA by ELISA

  11. Comparison of peptide mass mapping and electron capture dissociation as assays for histone posttranslational modifications

    Science.gov (United States)

    Zhang, Liwen; Freitas, Michael A.

    2004-05-01

    Posttranslational modifications of core histones play a critical role in the structure of chromatin and the regulation of gene activities. Improved techniques for determining these modification sites may lead to a better understanding of histone regulation at the molecular level. LC-MS peptide mass mapping was performed on pepsin, trypsin and Glu-C digests of bovine thymus H4 using a QqTOF instrument. The well established modification sites of H4 (acetylation of K8, 12, 16 and methylation of K20) were observed in addition to several recently discovered modifications including: methylation of K31, 44, 59 and acetylation of K20, 77, 79. For comparison, electron capture dissociation (ECD) was performed on intact H4 along with several peptides from enzymatic digestion. The results from the ECD experiments of histone H4 indicated the acetylation of K5, 12, 16, 31, 91 and the methylation of K20 and 59 in good agreement with the result from peptide mapping. The work is dedicated to Alan G. Marshall on his 60th birthday. His endeavors in the advancement of FT-ICR facilitated experiments reported herein.

  12. Diagnostic tests for amoebic liver abscess: comparison of enzyme - linked immunosorbent assay (Elisa and counterimmunoelectrophoresis (CIE

    Directory of Open Access Journals (Sweden)

    Marcos I. Restrepo

    1996-02-01

    Full Text Available The liver abscess is the most frequent extraintestinal complication of intestinal amoebiasis: its diagnosis is suggested by the clinical picture but it must be confirmed by paraclinic tests. Themost stringent diagnosis requires identification of E. histolytica. But this is possible only in a few cases. Serological tests greatly improve the diagnosis of this severe complication of amoebiasis. We compared the Enzyme Linfed Immunosorbent Assay and the Counterimmunoeletrophoresis techniques. Both techniques were used to detect amoebic antibodies in 50 control patients, 30 patients with liver abscess and 30 patients with intestinal amoebiasis. All the sera from control patients gave negative results iin both techniques. When analysing the sera from patients with intestinal amoebiasis, 10% of them were positive by ELISA but non by CIE. The sera of patients with liver abscess, we found that 90% were positive by the ELISA method and 66.6% by the CIE technique. In patients with amoebic liver abscess, the results showed that the ELISA was more sensitive than the CIE, as it presented a higher sensitivity (100% than that of the CIE technique (66%.O abscesso hepático é a complicação mais freqüente da amebíase intestinal: o seu diagnótico sugere-se pelo quadro clínico, mas é confirmado pelos estudos paraclínicos. Para confirmar o diagnóstico precisa-se identificar a E. histolytica, o que é apenas possível em muito poucos casos. As provas sorolôgicas melhoram notadamente o diagnóstico das complicações severas da amebíase. Em nosso estudo comparamos o teste de ELISA e a Contraimunoeletroforese (CIE. Ambas as técnicas foram utilizadas para detectar anticorpos contra ameba em 50 pacientes sem amebíase, 30 pacientes com abscesso hepático e 30 com amebíase intestinal. Todos os soros dos pacientes sem amebíase foram negativos por ambas as técnicas. Quando analisamos os soros dos pacientes com amebíase intestinal, encontrou-se que 10% destes

  13. Comparison of four molecular assays for the detection of Tembusu virus.

    Science.gov (United States)

    Tang, Yi; Yeh, Yin-Ting; Chen, Hao; Yu, Chunmei; Gao, Xuhui; Diao, Youxiang

    2015-10-01

    Tembusu virus (TMUV) belongs to the genus Flavivirus that may cause severe egg drop in ducks. In order to evaluate the most efficient TMUV detection method, the performances of a conventional RT-PCR (C-RT-PCR), a semi-nested PCR (SN-RT-PCR), a reverse-transcriptase real-time quantitative PCR (Q-RT-PCR), and a reverse-transcription loop-mediated isothermal amplification (RT-LAMP) targeting the TMUV virus-specific NS5 gene were examined. In order to compare the sensitivity of these four techniques, two templates were used: (1) plasmid DNA that contained a partial region of the NS5 gene and (2) genomic RNA from TMUV-positive cell culture supernatants. The sensitivities using plasmid DNA detection by C-RT-PCR, SN-RT-PCR, Q-RT-PCR, and RT-LAMP were 2 × 10(4) copies/μL, 20 copies/μL, 2 copies/μL, and 20 copies/μL, respectively. The sensitivities using genomic RNA for the C-RT-PCR, SN-RT-PCR, Q-RT-PCR, and RT-LAMP were 100 pg/tube, 100, 10, and 100 fg/tube, respectively. All evaluated assays were specific for TMUV detection. The TMUV-specific RNA was detected in cloacal swabs from experimentally infected ducks using these four methods with different rates (52-92%), but not in the control (non-inoculated) samples. The sensitivities of RT-PCR, SN-RT-PCR, Q-RT-PCR, and RT-LAMP performed with cloacal swabs collected from suspected TMUV infected ducks within 2 weeks of severe egg-drop were 38/69 (55.1%), 52/69 (75.4%), 57/69 (82.6%), and 55/69 (79.7%), respectively. In conclusion, both RT-LAMP and Q-RT-PCR can provide a rapid diagnosis of TMUV infection, but RT-LAMP is more useful in TMUV field situations or poorly equipped laboratories.

  14. Obesity in occupational groups of Western Siberia: comparison with representative national data

    Directory of Open Access Journals (Sweden)

    S A Maksimov

    2013-03-01

    Full Text Available The aim of this study was to compare obesity prevalences in the occupational groups of Western Siberia with the national data. Materials and methods: We performed a single-step cross-sectional study enrolling 4472 employees of 14 occupational groups from Western Siberian institutions and enterprises. Obesity was considered to be present if the body mass index was >30.0 kg/m2; sex, age and education data were obtained with questionnaires. Age-adjusted obesity prevalence in the occupational groups (separately for men and women was compared with the national data with calculation of odds ratio, attributable risk and 95% confidence interval. Results: Among women the prevalence of obesity was lower in teachers compared with the national data (OR=0.45; 95% CI: 0.31–0.66. Higher obesity prevalence was observed among operating personnel and technical workers (OR=1.69; 95% CI: 1.37–2.09 as well as metallurgy equipment operators (OR=1.65; 95% CI: 1.17–2.31. Among males higher obesity prevalence was registered in top-managers (OR=2.53; 95% CI: 1.80–3.55, operating personnel and technical workers (OR=2.03; 95% CI: 1.59–2.58, civil servants (OR=1.75; 95% CI: 1.27–2.40, and mechanics (OR=1.37; 95% CI: 1.08–1.73. Moreover, in women university education (higher percentage of employees having graduated from a higher professional institution led to less obesity prevalence. In males no such tendencies were observed. Conclusions: The study allowed to identify the occupational groups of Western Siberia with higher obesity prevalence and to demonstrate the impact of sex and education level on this parameter. The obtained data can make a theoretical and practical basis for primary and secondary prevention of obesity in the workplace.

  15. Comparison between Polish and Western European fish consumers in their attitudinal and behavioural patterns

    DEFF Research Database (Denmark)

    Pieniak, Zuzanna; Verbeke, Wim; Brunsø, Karen;

    2009-01-01

    The objective of this paper is to investigate consumer attitudes and behavioural patterns related to fish consumption in Poland and four Western European countries (Belgium, Denmark, the Netherlands, and Spain). A quantitative cross-sectional consumer survey was carried out and a total sample...... of 4786 respondents, representative within each country for age and region (n=800-1100 respondents per country) was obtained. Although Polish consumers have the most positive general attitudes toward fish and the strongest belief that eating fish is healthy and safe, their intention and fish consumption...

  16. Identification, expression profiling and fluorescence-based binding assays of a chemosensory protein gene from the Western flower thrips, Frankliniella occidentalis.

    Directory of Open Access Journals (Sweden)

    Zhi-Ke Zhang

    Full Text Available Using RT-PCR and RACE-PCR strategies, we cloned and identified a new chemosensory protein (FoccCSP from the Western flower thrips, Frankliniella occidentalis, a species for which no chemosensory protein (CSP has yet been identified. The FoccCSP gene contains a 387 bp open-reading frame encoding a putative protein of 128 amino acids with a molecular weight of 14.51 kDa and an isoelectric point of 5.41. The deduced amino acid sequence contains a putative signal peptide of 19 amino acid residues at the N-terminus, as well as the typical four-cysteine signature found in other insect CSPs. As FoccCSP is from a different order of insect than other known CSPs, the GenBank FoccCSP homolog showed only 31-50% sequence identity with them. A neighbor-joining tree was constructed and revealed that FoccCSP is in a group with CSPs from Homopteran insects (e.g., AgosCSP4, AgosCSP10, ApisCSP, and NlugCSP9, suggesting that these genes likely developed from a common ancestral gene. The FoccCSP gene expression profile of different tissues and development stages was measured by quantitative real-time PCR. The results of this analysis revealed this gene is predominantly expressed in the antennae and also highly expressed in the first instar nymph, suggesting a function for FoccCSP in olfactory reception and in particular life activities during the first instar nymph stage. We expressed recombinant FoccCSP protein in a prokaryotic expression system and purified FoccCSP protein by affinity chromatography using a Ni-NTA-Sepharose column. Using N-phenyl-1-naphthylamine (1-NPN as a fluorescent probe in fluorescence-based competitive binding assay, we determined the binding affinities of 19 volatile substances for FoccCSP protein. This analysis revealed that anisic aldehyde, geraniol and methyl salicylate have high binding affinities for FoccCSP, with KD values of 10.50, 15.35 and 35.24 μM, respectively. Thus, our study indicates that FoccCSP may play an important role in

  17. Comparison of microbiological assay and HPLC-UV for determination of fluconazole in capsules

    Directory of Open Access Journals (Sweden)

    Kelly Marques Queiroz

    2009-12-01

    Full Text Available The development of a specific agar diffusion bioassay for the quantitative determination of fluconazole formulated in capsules was carried out using a strain of Candida albicans ATCC 18804 as the test organism. A prospective validation of the method showed adequate linearity (r²=0.9995, precision (R.S.D. = 4.0% for intra-day and 4.5% for inter-day precision and accuracy (mean recovery = 102.9%. High performance liquid chromatography was chosen as a comparison method for the fluconazole determination. The contents of fluconazole determined by both methods, for four capsule samples, showed a strong correlation, confirmed by Pearson's correlation coefficient value (r = 0.9884. The bioassay is a suitable method for both research and pharmaceutical industry laboratories.Este trabalho visou ao desenvolvimento e validação de um método microbiológico por difusão em ágar para quantificação de fluconazol em cápsulas utilizando o isolado Candida albicans ATCC 18804 como reagente biológico. O método foi validado e foi verificada linearidade (r²=0,9995, precisão (D.P.R. = 4.0% para precisão intra-dia e 4,5% para precisão inter-dia e exatidão (recuperação média = 102,9%. Concomitantemente, foi realizado o doseamento de fluconazol nas cápsulas por meio de cromatografia líquida de alta eficiência. Os teores encontrados por ambos os métodos demonstraram alta correlação, confirmada pelo Coeficiente de Correlação de Pearson (r = 0,9884. O ensaio microbiológico desenvolvido pode ser considerado ferramenta valiosa tanto para a pesquisa científica quanto para a rotina da indústria farmacêutica.

  18. Comparison of climatic trends and variability among glacierized environments in the Western Himalayas

    Science.gov (United States)

    Dimri, A. P.; Immerzeel, W. W.; Salzmann, N.; Thayyen, R. J.

    2017-09-01

    The climate and hydrology of the Western Himalayas is complex and a function of snow and glacier melt, land use, topography, and Indian summer and winter monsoon dynamics. Improving our knowledge about these processes is important from societal and agricultural points of view. In this study, an observational analysis is carried out to assess the changing climatic trends and the associated interannual variability in winter temperature and precipitation at three glacierized regions of Western Himalayas having distinctly different sub-regional characteristics. In situ observations of 23 years (1985-2007) are used. These observations are passed through rigorous statistical quality control checks. Results show higher interannual variability with increasing temperature trends in the glacierized regions of the Siachen (Karakoram Range) and Chotasigri (Great Himalayan Range). Karakoram Range has higher warming trends than the Great Himalayan Range. In case of precipitation, an overall decrease in precipitation is observed with contrasting trends in the last decade. Nino3.4 index is positively correlated with winter precipitation with similar interannual variability. In addition, at Siachen temperature and precipitation show strong negative correlation, and precipitation to spell length correlation is opposite at Siachen and Chotasigri.

  19. Potential new production in two upwelling regions of the western Arabian Sea: Estimation and comparison

    Science.gov (United States)

    Liao, Xiaomei; Zhan, Haigang; Du, Yan

    2016-07-01

    Using satellite-derived and in situ data, the wind-driven potential new production (nitrate supply) for the 300 km wide coastal band in two upwelling regions of the western Arabian Sea (AS) during the southwest monsoon is estimated. The upward nitrate flux to the euphotic zone is generally based on the physical processes of coastal transport (Ekman transport and geostrophic transport) and offshore Ekman pumping. The coastal geostrophic current in the western AS influences the upwelling intensity and latitudinal distributions of nitrate supply. The Oman and Somalia upwelling regions have similar level of potential new production (nitrate supply) during the summer monsoon, while the satellite estimates of primary production off Oman are 2 times greater than those off Somalia. The much higher potential f-ratio in the Somalia upwelling region indicates that the primary production could be limited by availability of other macronutrients (e.g., silicate). The correlation analysis of the primary production and the aerosol optical thickness shows that the Oman upwelling region displays a stronger coupling between the atmospheric deposition and the phytoplankton abundance. The high summertime dust levels in the atmosphere are suggested to contribute to the high primary production in the Oman upwelling region.

  20. [Use of procalcitonine in intensive care units: comparison of semi quantitative PCT-Q Brahms assay with automated PCT-Kryptor assay].

    Science.gov (United States)

    Schuch, Géraldine; Duc-Marchand, Catherine; Venet, Cyrille; Mann, Hubert; Tixier, Anne; Bionda, Clara

    2011-01-01

    Procalcitonine (PCT) is recognized as a major and specific biomarker in diagnosis of bacterial infection. Used early in sepsis, it allows immediate administration of antibiotics and monitoring its effectiveness. Confronted on systemic inflammation response syndrom (SIRS), physicians must react quickly and effectively to evaluate bacterial infection and sepsis. The objective of this study was to compare analytical and clinical performances of semi-quantitative PCT-Q assay (Brahms) with quantitative and automated assay such on Kryptor (Brahms). Fifty blood samples of intensive care patients were compared. The analytical performance observed with PCT-Q assay is accurate: linear ratio kappa of 0.912 (95% CI 0.61, 0.97) and a good correlation between these techniques (p < 0.0001) (MedCalc software) were observed. Three discordances were observed and confirm the difficulties of reading for values close to 0.5 ng/mL. For these patients, PCT result showed its interest to discriminate local infection of a sepsis, to stop antibiotherapy with broad spectrum and to consolidate a therapeutic effectiveness in multi-visceral failure context. The semi-quantitative assay seems adapted for a fast and reliable evaluation of PCT in a general-purpose laboratory, not requiring neither dedicated analyzer, nor complex technicality but a control of the visual evaluation of results. It could be used for diagnosis of sepsis without monitoring precisely therapeutic follow-up.

  1. Comparison of a multiplex flow cytometric assay with enzyme-linked immunosorbent assay for auantitation of antibodies to tetanus, diphtheria, and Haemophilus influenzae Type b.

    Science.gov (United States)

    Pickering, Jerry W; Martins, Thomas B; Schroder, M Carl; Hill, Harry R

    2002-07-01

    We developed a multiplexed indirect immunofluorescence assay for antibodies to Haemophilus influenza type b (Hib) polysaccharide and the toxoids of Clostridium tetani (Tet) and Corynebacterium diphtheriae (Dip) based on the Luminex multiple-analyte profiling system. A pooled serum standard was calibrated against World Health Organization standards for Dip and Tet and an international standard for Hib. The multiplexed Luminex assay was compared to individual enzyme-linked immunosorbent assays (ELISAs) for the same analytes. By both methods, 75 (92.6%) of 81 of random serum samples had protective levels of antibody to Tet (> or = 0.1 IU/ml). For Dip, 81.5% of the samples had protective antibody levels (> or = 0.1 IU/ml) by ELISA and 80.2% had protective antibody levels by Luminex. Protective levels (> or = 1.0 microg/ml) of antibody to Hib were found in 45.0% of the samples tested by ELISA and in 39.0% of the samples tested by Luminex. The correlations (R(2)) between ELISA and Luminex of the 81 samples were 0.96, 0.96, and 0.91 for Tet, Dip, and Hib, respectively. There was also similar agreement between Luminex and ELISA for sera collected before and 1 month after Tet, Dip, and Hib vaccine administration. Both methods detected strong postvaccination responses. The Luminex method is an attractive alternative to ELISA since it reduces labor and reagent costs, as well as assay time.

  2. Comparison of two enzyme-linked immunosorbent assays and one rapid immunoblot assay for detection of herpes simplex virus type 2-specific antibodies in serum

    NARCIS (Netherlands)

    J. Groen (Jan); G. van Dijk (Grietje); H.G.M. Niesters (Bert); W.I. van der Meijden (Willem); A.D.M.E. Osterhaus (Albert)

    1998-01-01

    textabstractThe sensitivities and specificities of three immunoassays for the detection of herpes simplex virus type 2 (HSV-2)-specific immunoglobulin G antibodies in serum, including the one-strip rapid immunoblot assay (RIBA; Chiron Corporation) and two indirect enzyme

  3. Regional Conflicts in the Western Balkans and the Caucasus Revisited: Comparison of Kosovo to South Ossetia and Abkhazia

    Directory of Open Access Journals (Sweden)

    Vladimir Đorđević

    2010-04-01

    Full Text Available One of the things that the Western Balkans and the Caucasus have in common is an extremely challenging legacy of the past. The dissolution of two multinational states – the Soviet Union and Socialist Yugoslavia in the beginning of 1990s – led to ethno-nationalist conflicts on a large scale. While the Yugoslav crisis ended in 1999 after the FRY was bombed by NATO during its Kosovo campaign, the Caucasus still remains a conflict-ridden region where Russian and Western influences keep colliding. The purpose of this article is to present an analytical comparison of the three respective regional conflicts – Kosovo, Georgia and South Ossetia – by enumerating and analyzing similarities and differences between them, as this proves to be one of current and more intriguing issues of the contemporary international political scene. The article aims at providing answers to two different issues: Did Kosovo’s independence influence the establishment of a specific political pattern applicable to other disputed regions; and to what degree are the cases in question comparable to each other?

  4. Electrochemical Measurement of the β-Galactosidase Reporter from Live Cells: A Comparison to the Miller Assay.

    Science.gov (United States)

    Tschirhart, Tanya; Zhou, Xinyi Y; Ueda, Hana; Tsao, Chen-Yu; Kim, Eunkyoung; Payne, Gregory F; Bentley, William E

    2016-01-15

    In order to match our ability to conceive of and construct cells with enhanced function, we must concomitantly develop facile, real-time methods for elucidating performance. With these, new designs can be tested in silico and steps in construction incrementally validated. Electrochemical monitoring offers the above advantages largely because signal transduction stems from direct electron transfer, allowing for potentially quicker and more integrated measurements. One of the most common genetic reporters, β-galactosidase, can be measured both spectrophotometrically (Miller assay) and electrochemically. However, since the relationship between the two is not well understood, the electrochemical methods have not yet garnered the attention of biologists. With the aim of demonstrating the utility of an electrochemical measurement to the synthetic biology community, we created a genetic construct that interprets and reports (with β-galactosidase) on the concentration of the bacterial quorum sensing molecule autoinducer-2. In this work, we provide a correlation between electrochemical measurements and Miller Units. We show that the electrochemical assay works with both lysed and whole cells, allowing for the prediction of one from the other, and for continuous monitoring of cell response. We further present a conceptually simple and generalized mathematical model for cell-based β-galactosidase reporter systems that could aid in building and predicting a variety of synthetic biology constructs. This first-ever in-depth comparison and analysis aims to facilitate the use of electrochemical real-time monitoring in the field of synthetic biology as well as to facilitate the creation of constructs that can more easily communicate information to electronic systems.

  5. Comparison of Terrestrial Isopod (Isopoda, Oniscidea Assemblages from Two Types of Forests from North Western Romania

    Directory of Open Access Journals (Sweden)

    Sára Ferenţi

    2012-12-01

    Full Text Available In 2008 we compared the terrestrial isopod assemblages from two different habitats, a beech forest and a mixed beech and spruce forest, from north western Romania (Huta Certeze locality. The samples were taken from April to September using pitfall traps. We identified a total of 7 species: Ligidium germanicum, Trichoniscus sp., Hyloniscus transsilvanicus, Protracheoniscus politus, Porcellium collicola, Trachelipus difficilis and Porcellio scaber. A greater diversity and species richness were noticed in the beech forest. The poverty of species in the mixed forest was a consequence of the forest type, the anthropogenic impact and the dry environment. High surface activity of individuals was noticed in the summer months. Even if the species compositions of the two compared isopod assemblages were not identical, there weren’t statistically significant differences between them.

  6. Comparison of China-US Engineering Ethics Educations in Sino-Western Philosophies of Technology.

    Science.gov (United States)

    Cao, Gui Hong

    2015-12-01

    Ethics education has become essential in modern engineering. Ethics education in engineering has been increasingly implemented worldwide. It can improve ethical behaviors in technology and engineering design under the guidance of the philosophy of technology. Hence, this study aims to compare China-US engineering ethics education in Sino-Western philosophies of technology by using literature studies, online surveys, observational researches, textual analyses, and comparative methods. In my original theoretical framework and model of input and output for education, six primary variables emerge in the pedagogy: disciplinary statuses, educational goals, instructional contents, didactic models, teaching methods, and edificatory effects. I focus on the similarities and differences of engineering ethics educations between China and the U.S. in Chinese and Western philosophies of technology. In the field of engineering, the U.S. tends toward applied ethics training, whereas China inclines toward practical moral education. The U.S. is the leader, particularly in the amount of money invested and engineering results. China has quickened its pace, focusing specifically on engineering labor input and output. Engineering ethics is a multiplayer game effected at various levels among (a) lower level technicians and engineers, engineering associations, and stockholders; (b) middle ranking engineering ethics education, the ministry of education, the academy of engineering, and the philosophy of technology; and (c) top national and international technological policies. I propose that professional engineering ethics education can play many important roles in reforming engineering social responsibility by international cooperation in societies that are becoming increasingly reliant on engineered devices and systems. Significantly, my proposals contribute to improving engineering ethics education and better-solving engineering ethics issues, thereby maximizing engineering

  7. Vascular trauma in Western Australia: a comparison of two study periods over 15 years.

    Science.gov (United States)

    Friend, Jikol; Rao, Sudhakar; Sieunarine, Kishore; Woodroof, Paul

    2016-03-01

    Royal Perth Hospital (RPH) has become Western Australia's only designated adult major trauma facility since a previous study of vascular trauma was conducted in 2001 at the same facility. The aim of this study is to identify changes in vascular trauma patterns over the two study periods and compare these changes with international literature. All individuals presenting to RPH between January 2000 and December 2010 with vascular injury were identified from a prospective trauma database for this descriptive study. Injuries were classified using the Abbreviated Injury Score (AIS). The incidence of vascular trauma as a percentage of total trauma increased over the two study periods. The current 10-year study included 45 164 patients on the trauma database, of which 1205 patients (2.6%) sustained 1335 vascular injuries, an increase from 1% in the previous 5-year study at the same facility. Males aged 20-29 years were more frequently injured. Blunt trauma occurred more frequently than penetrating. The extremities, particularly the upper limbs were most commonly injured. The most common causes of injury for each region were as follows; motorbike crash (MBC), motor vehicle crash (MVC) and stabbing (neck, thorax and abdomen), MBC and MVC (lower limb) and piercing injuries (upper limb). Injury Severity Score (ISS) and mortality 43% (32 of 75) were highest for thoracic injuries, particularly thoracic aorta injury. Mortality rate has decreased. Vascular injuries in Western Australia are increasing. MVC are the most common cause of life threatening injury. Road safety interventions targeting young males are likely to reduce trauma. © 2013 Royal Australasian College of Surgeons.

  8. Comparison of a PCR serotyping assay, Check&Trace assay for Salmonella, and Luminex Salmonella serotyping assay for the characterization of Salmonella enterica identified from fresh and naturally contaminated cilantro.

    Science.gov (United States)

    Jean-Gilles Beaubrun, J; Ewing, L; Jarvis, K; Dudley, K; Grim, C; Gopinath, G; Flamer, M-L; Auguste, W; Jayaram, A; Elmore, J; Lamont, M; McGrath, T; Hanes, D E

    2014-09-01

    Salmonella enterica isolated from fresh cilantro samples collected through the USDA/AMS Microbiological Data Program (MDP) were used to compare a PCR serotyping assay against the Check&Trace assay and the Luminex (BioPlex) Salmonella serotyping assay. The study was conducted to evaluate the effectiveness of the three methods for serotyping Salmonella from both enrichment broth cultures and pure Salmonella cultures. In this investigation, Salmonella spp. serotyping was conducted using 24 h enrichment broth cultures and pure Salmonella cultures from cilantro samples, with the PCR serotyping assay. Conversely, the Check&Trace and Luminex for Salmonella assays required pure cultures for Salmonella serotyping. The cilantro samples contained S. enterica serovar Montevideo, Newport, Saintpaul, and Tennessee, identified by the PCR serotyping assay and Check&Trace for Salmonella, but the Luminex assay only identified two of the four serotypes of the cilantro samples. The anticipated impact from this study is that the PCR serotyping assay provides a time- and cost-effective means for screening, identifying and serotyping Salmonella using DNA extracted from 24 h enrichment cilantro samples.

  9. Improving validation methods for molecular diagnostics: application of Bland-Altman, Deming and simple linear regression analyses in assay comparison and evaluation for next-generation sequencing.

    Science.gov (United States)

    Misyura, Maksym; Sukhai, Mahadeo A; Kulasignam, Vathany; Zhang, Tong; Kamel-Reid, Suzanne; Stockley, Tracy L

    2017-07-26

    A standard approach in test evaluation is to compare results of the assay in validation to results from previously validated methods. For quantitative molecular diagnostic assays, comparison of test values is often performed using simple linear regression and the coefficient of determination (R(2)), using R(2) as the primary metric of assay agreement. However, the use of R(2) alone does not adequately quantify constant or proportional errors required for optimal test evaluation. More extensive statistical approaches, such as Bland-Altman and expanded interpretation of linear regression methods, can be used to more thoroughly compare data from quantitative molecular assays. We present the application of Bland-Altman and linear regression statistical methods to evaluate quantitative outputs from next-generation sequencing assays (NGS). NGS-derived data sets from assay validation experiments were used to demonstrate the utility of the statistical methods. Both Bland-Altman and linear regression were able to detect the presence and magnitude of constant and proportional error in quantitative values of NGS data. Deming linear regression was used in the context of assay comparison studies, while simple linear regression was used to analyse serial dilution data. Bland-Altman statistical approach was also adapted to quantify assay accuracy, including constant and proportional errors, and precision where theoretical and empirical values were known. The complementary application of the statistical methods described in this manuscript enables more extensive evaluation of performance characteristics of quantitative molecular assays, prior to implementation in the clinical molecular laboratory. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

  10. Comparison of Brucella immunoglobulin M and G flow assays with serum agglutination and 2-mercaptoethanol tests in the diagnosis of brucellosis

    NARCIS (Netherlands)

    A. Zeytinoglu; A. Turhan; I. Altuglu; A. Bilgic; T.H. Abdoel; H.L. Smits

    2006-01-01

    The diagnostic value of Brucella IgM/IgG flow assays was evaluated in comparison with serum agglutination and 2-mercaptoethanol tests by testing a selection of serum samples submitted to the laboratory because of clinical suspicion of brucellosis. All 39 admission and 11 follow-up samples that agglu

  11. A Comparison of the Human Buccal Cell Assay and the Pollen Abortion Assay in Assessing Genotoxicity in an Urban-Rural Gradient

    Directory of Open Access Journals (Sweden)

    Alan da Silveira Fleck

    2014-08-01

    Full Text Available Air pollution is exacerbated near heavy traffic roads in cities. Air pollution concentration and composition vary by region and depend on urban-rural gradients. The aim of this study was to evaluate the distribution of air pollution in areas of varying population densities and to compare plant biomonitoring with an established biomarker of human exposure to traffic-related air pollution in children. The areas of study were selected near a major street in 3 different regions. Areas A, B and C represent high, intermediate and low population densities, respectively. Micronucleus assay, an established biomarker of human exposure, was performed in children from these areas. For a plant biomonitoring assay, the pollen abortion assay was performed on Bauhinia variegata in these areas. NO2 and O3 concentrations were determined by passive sampling. We report here that the pollen abortion frequency in Bauhinia variegata is correlated with NO2 concentration (P = 0.004 and is strongly associated with vehicular flow and population density in the studied areas. Micronuclei frequency in buccal cells of children was higher in the regions with more degree of urbanization (P < 0.001 following the same pattern of O3 concentrations (P = 0.030. In conclusion, our results demonstrate that high concentrations of air pollutants in Porto Alegre are related to both human and plant genotoxicity. Areas with different concentration of pollutants demonstrated to have an urbanization gradient dependent pattern which also reflected on genotoxic damage among these areas.

  12. A Comparison of the Human Buccal Cell Assay and the Pollen Abortion Assay in Assessing Genotoxicity in an Urban-Rural Gradient

    Science.gov (United States)

    Fleck, Alan da Silveira; Vieira, Mariana; Amantéa, Sergio Luís; Rhoden, Claudia Ramos

    2014-01-01

    Air pollution is exacerbated near heavy traffic roads in cities. Air pollution concentration and composition vary by region and depend on urban-rural gradients. The aim of this study was to evaluate the distribution of air pollution in areas of varying population densities and to compare plant biomonitoring with an established biomarker of human exposure to traffic-related air pollution in children. The areas of study were selected near a major street in 3 different regions. Areas A, B and C represent high, intermediate and low population densities, respectively. Micronucleus assay, an established biomarker of human exposure, was performed in children from these areas. For a plant biomonitoring assay, the pollen abortion assay was performed on Bauhinia variegata in these areas. NO2 and O3 concentrations were determined by passive sampling. We report here that the pollen abortion frequency in Bauhinia variegata is correlated with NO2 concentration (P = 0.004) and is strongly associated with vehicular flow and population density in the studied areas. Micronuclei frequency in buccal cells of children was higher in the regions with more degree of urbanization (P < 0.001) following the same pattern of O3 concentrations (P = 0.030). In conclusion, our results demonstrate that high concentrations of air pollutants in Porto Alegre are related to both human and plant genotoxicity. Areas with different concentration of pollutants demonstrated to have an urbanization gradient dependent pattern which also reflected on genotoxic damage among these areas. PMID:25166920

  13. A comparison of the human buccal cell assay and the pollen abortion assay in assessing genotoxicity in an urban-rural gradient.

    Science.gov (United States)

    Fleck, Alan da Silveira; Vieira, Mariana; Amantéa, Sergio Luís; Rhoden, Claudia Ramos

    2014-08-27

    Air pollution is exacerbated near heavy traffic roads in cities. Air pollution concentration and composition vary by region and depend on urban-rural gradients. The aim of this study was to evaluate the distribution of air pollution in areas of varying population densities and to compare plant biomonitoring with an established biomarker of human exposure to traffic-related air pollution in children. The areas of study were selected near a major street in 3 different regions. Areas A, B and C represent high, intermediate and low population densities, respectively. Micronucleus assay, an established biomarker of human exposure, was performed in children from these areas. For a plant biomonitoring assay, the pollen abortion assay was performed on Bauhinia variegata in these areas. NO2 and O3 concentrations were determined by passive sampling. We report here that the pollen abortion frequency in Bauhinia variegata is correlated with NO2 concentration (P = 0.004) and is strongly associated with vehicular flow and population density in the studied areas. Micronuclei frequency in buccal cells of children was higher in the regions with more degree of urbanization (P < 0.001) following the same pattern of O3 concentrations (P = 0.030). In conclusion, our results demonstrate that high concentrations of air pollutants in Porto Alegre are related to both human and plant genotoxicity. Areas with different concentration of pollutants demonstrated to have an urbanization gradient dependent pattern which also reflected on genotoxic damage among these areas.

  14. Regional boreal summer intraseasonal oscillation over Indian Ocean and Western Pacific: comparison and predictability study

    Science.gov (United States)

    Lee, Sun-Seon; Wang, Bin

    2016-04-01

    The boreal summer intraseasonal oscillation (BSISO) has two major activity centers, the northern Indian Ocean and tropical Western North Pacific, which dominate the monsoon intraseasonal variability over South Asia and East Asia, respectively. The spatial-temporal structures of BSISO over the Indian Ocean (10°S-30°N, 60°-105°E) (IOISO) and Western Pacific (10°S-30°N, 105°-150°E) (WPISO) are examined by corresponding the leading modes of daily OLR and 850-hPa zonal wind (U850). The IOISO features a northeastward propagation with a 30-45 days energy peak and the first principal component (PC1) has maximum variance in May, while the WPISO propagates northward with a broad spectral peak on 10-60 days and the PC1 has maximum variance in August. Because of the large regional differences, two regional indices, the IOISO index and WPISO index, are defined by their corresponding first two leading PCs. The combined IOISO-WPISO index captures about 30 % (10 %) of U850 (OLR) daily variance over the entire IO-WP region (10°S-30°N, 60°-150°E), which doubles that captured by the Madden-Julian Oscillation (MJO) index (Wheeler and Hendon 2004) and is 50 % higher than that captured by the BSISO index (Lee et al. 2013). The combined index also shows superior performance in representing biweekly and pentad-mean variations in the Asian-Pacific summer monsoon region (north of 10°N). The predictability/prediction skill and simulated principal modes of two regional BSISO indices are explored by using data derived from the Intraseasonal Variability Hindcast Experiment project. The major regional modes are reasonably well captured, but the forecasted fractional variances of the leading modes and variability center's locations exhibit significant deficiencies. The multi-model mean estimate of the predictability is 40-45 days for the IOISO index, whereas 33-37 days for the WPISO index. The less predictable WPISO is likely due to the existence of its significant biweekly component

  15. Comparison between measurements of black carbon, charcoal and associated nutrients in western Amazonan soils

    Science.gov (United States)

    Zimmerman, A. R.; McMichael, C.; Hanlon, C.; Bush, M. B.

    2011-12-01

    To construct fire and climate history and human occupation records from soils and lake sediment profiles, climatologists and anthropologists have traditionally measured charcoal abundances by microscopic image analysis. In contrast, geochemists have developed methods of black carbon (BC) quantification using chemical extraction. We compared charcoal (>0.5 mm particle size) versus BC (measured via the CTO-340 method of Kuhlbusch, 1995) in multiple soil profiles from four western Amazon regions with evidence of pre-Columbian occupation. A secondary goal of this project was to understand the relative influence of climate and humans in the fire and ecological history of the Amazon. BC concentration in soils of the Amazon varied widely from an average of 0.5 mg g 1 in cores around Lake Gentry (southeastern Peru) to 5.5 mg g 1 around Lake Ayauchi (southeastern Ecuador), corresponding to the evidence of greater land use around the latter. Surprising, BC concentrations in habitation horizon soils at Quistococha, near Iquitos, Peru were similar to Lake Gentry, averaging about 0.6 mg g 1. However, BC as a percent of soil organic carbon (SOC) was much more uniform with an average of 12.0, 13.3, 14.6, and 13.0% in Quistococha, Gentry, Ayauchi, and Los Amigos (central-eastern Peru) soils, respectively, suggesting that the same processes that concentrate SOC also concentrate BC. BC may act to protect SOC via sorption or produce SOC via microbial community enhancement. These findings also show that BC is not regionally enriched as it might be were climate to be a predominant factor in BC production, and seem to track land use more closely. Charcoal and BC concentrations were linearly correlated in only about half the soil profiles and neither BC nor charcoal were consistently correlated with chemical anthropogenic indicators such as P or Ca within soil profiles or specific regions. However, there was a statistical covariance between each of these parameters suggesting that each

  16. Comparison of the Abbott RealTime CT new formulation assay with two other commercial assays for detection of wild-type and new variant strains of Chlamydia trachomatis

    DEFF Research Database (Denmark)

    Møller, Jens Kjølseth; Pedersen, Lisbeth Nørum; Persson, Kenneth

    2010-01-01

    In an analytical methods comparison study on clinical samples, the Abbott RealTime CT new formulation assay (m2000 real-time PCR) consisting of a duplex PCR targeting different parts of the cryptic plasmid in Chlamydia trachomatis was compared with version 2 of the Roche COBAS(R) TaqMan(R) CT assay...... comprising a duplex PCR for a target in the cryptic plasmid and the omp1 gene, and compared with the Gen-Probe APTIMA COMBO 2(R) assay (AC2) targeting the C. trachomatis 23S rRNA molecule. First-catch urine samples from Sweden were tested in Malmoe for C. trachomatis with the m2000 real-time PCR assay......, and with an in-house PCR for the new variant C. trachomatis strain with a deletion in the cryptic plasmid. Aliquots of the urine samples were sent to Aarhus, Denmark and further examined with the TaqMan CT and the AC2 assay. A positive prevalence of 9.1% (148/1,632 urine samples examined) was detected according...

  17. Comparison of outcomes obtained in murine local lymph node assays using CBA/J or CBA/Ca mice.

    Science.gov (United States)

    Maeda, Yosuke; Hirosaki, Haruka; Yakata, Naoaki; Takeyoshi, Masahiro

    2016-08-01

    CBA/J and CBA/Ca mice are the recommended strains for local lymph node assays (LLNAs). Here, we report quantitative and qualitative comparisons between both mouse strains to provide useful information for the strain selection of sensitization testing. LLNA was conducted, in accordance with Organisation for Economic Co-operation and Development Test Guideline No. 429, with CBA/J and CBA/Ca mice using five chemicals including typical contact sensitizers and non-sensitizers: 2,4-dinitrochlorobenzene (DNCB), isoeugenol, α-hexylcinnamic aldehyde (HCA), propylene glycol (PG), and hexane; then outcomes were compared based on the raw data (disintegrations per minute, DPM), stimulation index (SI) values, EC3 values and positive/negative decisions. Although a significant difference was noted between DPM values derived from each strain of mice, SI values exhibited no considerable difference. The EC3 values for DNCB in CBA/J and CBA/Ca mice were 0.04 and 0.03, those for isoeugenol were 1.4 and 0.9, and those for HCA were 7.7 and 6.0, respectively. All EC values derived from each test system were almost equivalent and were within the range of acceptance criteria of the ICCVAM performance standard for LLNA. Positive/negative outcomes for all test chemicals were consistent. In conclusion, no considerable differences were observed in the final outcomes derived from CBA/J and CBA/Ca mice in LLNA. Copyright © 2015 John Wiley & Sons, Ltd.

  18. Risk factors for peripheral arterial disease in the tropics and its comparison with the western population

    Institute of Scientific and Technical Information of China (English)

    Myla Yacob; Edwin Stephen; Nupur Bit; Mazda Turel; David Sadhu; Sunil Agarwal

    2010-01-01

    Objective:To identify and compare the existence of similar and other risk factors in the perspective of an Indian population. Methods:It was designed as a case control study and was conducted in the Department of General and Vascular Surgery Unit 2 of Christian Medical College, Vellore, India between the periods July 2003 to June 2005. 100 patients with an ABPI<0.9 and 100 controls were studied. Results:Peripheral arterial disease (PAD) was found to be commoner among males (87%). While atherosclerosis was the commonest aetiology (54%), the incidence of Thromboangiitis Obliterans was also not uncommon (38%). Smoking was the main risk factor in the Indian context (83%) as compared to hypercholesterolemia (60%) in the West. The patients with atherosclerotic PAD were middle-aged and had concomitant diabetes (50%) and hypertension (30%). Conclusions:Peripheral arterial disease occurs in a relatively younger age group in India as compared to their Western counterparts. Thromboangiitis Obliterans was found to be a significant aetiology for arterial occlusive disease, with smoking as the primary risk factor followed by diabetes, hypertension and hypercholesterolemia.

  19. Quantity estimation and comparison in western lowland gorillas (Gorilla gorilla gorilla).

    Science.gov (United States)

    Vonk, Jennifer; Torgerson-White, Lauri; McGuire, Molly; Thueme, Melissa; Thomas, Jennifer; Beran, Michael J

    2014-05-01

    We investigated the quantity judgment abilities of two adult male western lowland gorillas (Gorilla gorilla gorilla) by presenting discrimination tasks on a touch-screen computer. Both gorillas chose the larger quantity of two arrays of dot stimuli. On some trials, the relative number of dots was congruent with the relative total area of the two arrays. On other trials, number of dots was incongruent with area. The gorillas were first tested with static dots, then with dots that moved within the arrays, and finally on a task where they were required to discriminate numerically larger subsets within arrays of moving dots. Both gorillas achieved above-chance performance on both congruent and incongruent trials with all tasks, indicating that they were able to use number as a cue even though ratio of number and area significantly controlled responding, suggesting that number was not the only relevant dimension that the gorillas used. The pattern of performance was similar to that found previously with monkeys and chimpanzees but had not previously been demonstrated in gorillas within a computerized test format, and with these kinds of visual stimuli.

  20. A comparison of tectonic ambient shear stress value in China with that in western USA

    Institute of Scientific and Technical Information of China (English)

    陈培善; 白彤霞; 李保昆

    2002-01-01

    A method is proposed to estimate average tectonic ambient shear stress value for a region. Thus the average stress values of 19 regions in western USA, and 43 regions (each region is 10((10() in Chinese mainland and its surroundings have been obtained. The data of 15 993 earthquakes are from the Internet Centroid Moment Tensor solution made by Harvard University from 1997 to 1999. The results demonstrate that there are highest average stress values in the regions of south California of USA and its off coast sea, reach to 12.0 MPa and 13.7 MPa respectively, then gradually decrease toward north, south, and east. The lowest value is 8.7 MPa and 63% of highest value. The average stress values in northern Xinjiang and in the Chayu region of Tibet are 17.2 and 12.9 MPa respectively. They are highest values in China and higher than USA(s. The average stress value in North China, Yunnan, Sichuan, Taiwan is similar to south California of USA. The average stress value in South-North seismic zone is about 13 MPa, a little higher than south California. The distribution of average stress value for two important regions provides basic data for geology. These results are useful to research earthquake activity background and attenuation relation of strong ground motion parameters (e.g. peak acceleration and response spectra).

  1. Comparison of gull-specific assays targeting 16S rRNA gene of Catellicoccus marimammalium and Streptococcus spp.

    Science.gov (United States)

    Gulls have been implicated as a source of fecal contamination in inland and coastal waters. Only one gull-specific assay is currently available (i.e., gull2 qPCR assay). This assay is based on the 16S rRNA gene of Catellicocclls marimammalium and has showed a high level of host-s...

  2. Comparison of two high-throughput assays for quantification of adenovirus type 5 neutralizing antibodies in a population of donors in China.

    Directory of Open Access Journals (Sweden)

    Qiang Liu

    Full Text Available BACKGROUND: The presence of various levels of Adenovirus serotype 5 neutralizing antibodies (Ad5NAb is thought to contribute to the inconsistent clinical results obtained from vaccination and gene therapy studies. Currently, two platforms based on high-throughput technology are available for Ad5NAb quantification, chemiluminescence- and fluorescence-based assays. The aim of this study was to compare the results of two assays in the seroepidemiology of Ad5NAb in a local population of donors. METHODOLOGY/PRINCIPAL FINDINGS: The fluorescence-based neutralizing antibody detection test (FRNT using recombinant Ad5-EGFP virus and the chemiluminescence-based neutralizing antibody test (CLNT using Ad5-Fluc were developed and standardized for detecting the presence of Ad5NAb in serum samples from the population of donors in Beijing and Anhui provinces, China. First, the overall percentage of people positive for Ad5NAb performed by CLNT was higher than that obtained by FRNT (85.4 vs 69.9%, p<0.001. There was an 84.5% concordance between the two assays for the 206 samples tested (144 positive in both assays and 30 negative in both assays. All 32 discordant sera were CLNT-positive/FRNT-negative and were confirmed positive by western blot. Secondly, for all 144 sera positive by both assays, the two assays showed high correlation (r = 0.94, p<0.001 and close agreement (mean difference: 0.395 log(10, 95% CI: -0.054 log(10 to 0.845 log(10. Finally, it was found by both assays that there was no significant difference observed for titer or prevalence by gender (p = 0.503 vs 0.818, for two assays; however, age range (p = 0.049 vs 0.010 and geographic origin (p = 0.007 vs 0.011 were correlated with Ad5NAb prevalence in northern regions of China. CONCLUSION: The CLNT assay was relatively more simple and had higher sensitivity than the FRNT assay for determining Ad5NAb titers. It is strongly suggested that the CLNT assay be used for future

  3. Comparison of quality control for trauma management between Western and Eastern European trauma center

    Directory of Open Access Journals (Sweden)

    Gambale Giorgio

    2008-11-01

    Full Text Available Abstract Background Quality control of trauma care is essential to define the effectiveness of trauma center and trauma system. To identify the troublesome issues of the system is the first step for validation of the focused customized solutions. This is a comparative study of two level I trauma centers in Italy and Romania and it has been designed to give an overview of the entire trauma care program adopted in these two countries. This study was aimed to use the results as the basis for recommending and planning changes in the two trauma systems for a better trauma care. Methods We retrospectively reviewed a total of 182 major trauma patients treated in the two hospitals included in the study, between January and June 2002. Every case was analyzed according to the recommended minimal audit filters for trauma quality assurance by The American College of Surgeons Committee on Trauma (ACSCOT. Results Satisfactory yields have been reached in both centers for the management of head and abdominal trauma, airway management, Emergency Department length of stay and early diagnosis and treatment. The main significant differences between the two centers were in the patients' transfers, the leadership of trauma team and the patients' outcome. The main concerns have been in the surgical treatment of fractures, the outcome and the lacking of documentation. Conclusion The analyzed hospitals are classified as Level I trauma center and are within the group of the highest quality level centers in their own countries. Nevertheless, both of them experience major lacks and for few audit filters do not reach the mmum standard requirements of ACS Audit Filters. The differences between the western and the eastern European center were slight. The parameters not reaching the minimum requirements are probably occurring even more often in suburban settings.

  4. Hypertension knowledge in urban elderly patients: comparison between adherents to traditional Chinese medicine and Western medicine

    Institute of Scientific and Technical Information of China (English)

    Jiangping Lin; Huining Lei; Fang Liu

    2008-01-01

    Objective To compare knowledge about hypertension between elderly Chinese urban patients with preferences for either traditional Chinese medicine (TCM) or Western medicine (WM).Methods Elderly (≥ 65 years old) patients with hypertension who prefer TCM treatment (n=112) or WM (n=126) were questioned about hypertension.Their answers were compared.Results Only 32.6% of participants correctly identified hypertension as a main risk factor of coronary heart disease and stroke,22.3% of patients answered that the main purpose of hypertension control was preventing cardiovascular disease.Other major reasons for these patients to seek medical treatment for their hypertension included:persuasion by physicians or their family members (21.6%),alleviating symptoms such as headache and dizziness (16.8%),lowering blood pressure without knowing specific reason (12.4%).The predictors for poor knowledge of hypertension were similar irrespective of preference for WM or TCM treatment,and included those with lower levels of education and older age.Television and newspaper (46.8%) were the most frequent sources of hypertension information for both groups.Among those who preferred TCM treatment,"TCM has fewer side effects than WM" and "TCM cures disease while WM only alleviates symptoms" were common beliefs held.Conclusion This study shows that knowledge of hypertension is similar among Chinese urban patients with preferences for either WM or TCM treatment and that misunderstandings about hypertension are common among the elderly patients.In order to control hypertension effectively,public health education is necessary.This should target those with a lower level of education and older age.

  5. A comparison of hydrothermal reservoirs of the Western United States. Topical Report 3

    Energy Technology Data Exchange (ETDEWEB)

    Meidav, H. Tsvi; Sanyal, Subir

    1976-12-01

    This report presents a portion of the results from a one-year feasibility study sponsored by the Electric Power Research Institute to assess the feasibility of constructing a 25 to 50 MWe geothermal power plant using low salinity hydrothermal fluids as the energy source. It contains the results of a comparative study of sixteen hydrothermal reservoirs in the US. The reservoirs were selected for comparison on the basis of available data, development potential, and representativeness of known hydrothermal reservoirs in the US. Six reservoir and fluid criteria were considered the most important in determining the development and power conversion potential: depth and lithology, reservoir temperature, tested flow rate per well, fluid chemistry, magnitude of the reserve and reinjection potential. These criteria were evaluated for each of the selected reservoirs.

  6. DNA-repair measurements by use of the modified comet assay: an inter-laboratory comparison within the European Comet Assay Validation Group (ECVAG).

    Science.gov (United States)

    Godschalk, Roger W L; Ersson, Clara; Riso, Patrizia; Porrini, Marisa; Langie, Sabine A S; van Schooten, Frederik-Jan; Azqueta, Amaya; Collins, Andrew R; Jones, George D D; Kwok, Rachel W L; Phillips, David H; Sozeri, Osman; Allione, Alessandra; Matullo, Giuseppe; Möller, Lennart; Forchhammer, Lykke; Loft, Steffen; Møller, Peter

    2013-09-18

    The measurement of DNA-repair activity by extracts from cells or tissues by means of the single-cell gel electrophoresis (comet) assay has a high potential to become widely used in biomonitoring studies. We assessed the inter-laboratory variation in reported values of DNA-repair activity on substrate cells that had been incubated with Ro19-8022 plus light to generate oxidatively damaged DNA. Eight laboratories assessed the DNA-repair activity of three cell lines (i.e. one epithelial and two fibroblast cell lines), starting with cell pellets or with cell extracts provided by the coordinating laboratory. There was a large inter-laboratory variation, as evidenced by the range in the mean level of repair incisions between the laboratory with the lowest (0.002incisions/10(6)bp) and highest (0.988incisions/10(6)bp) incision activity. Nevertheless, six out of eight laboratories reported the same cell line as having the highest level of DNA-repair activity. The two laboratories that reported discordant results (with another cell line having the highest level of DNA-repair activity) were those that reported to have little experience with the modified comet assay to assess DNA repair. The laboratories were also less consistent in ordering the repair activity of the other two cell lines, probably because the DNA-repair activity by extracts from these cell lines were very similar (on average approximately 60-65% of the cell line with the highest repair capacity). A significant correlation was observed between the repair activity found in the provided and the self-made cell extracts (r=0.71, Pcomet assay for base-excision repair.

  7. Aquaporin-4 autoantibodies in neuromyelitis optica spectrum disorders: comparison between tissue-based and cell-based indirect immunofluorescence assays

    Directory of Open Access Journals (Sweden)

    Chan Koon H

    2010-09-01

    Full Text Available Abstract Background Neuromyelitis optica spectrum disorders (NMOSD are severe central nervous system inflammatory demyelinating disorders (CNS IDD characterized by monophasic or relapsing, longitudinally extensive transverse myelitis (LETM and/or optic neuritis (ON. A significant proportion of NMOSD patients are seropositive for aquaporin-4 (AQP4 autoantibodies. We compared the AQP4 autoantibody detection rates of tissue-based indirect immunofluorescence assay (IIFA and cell-based IIFA. Methods Serum of Chinese CNS IDD patients were assayed for AQP4 autoantibodies by tissue-based IIFA using monkey cerebellum and cell-based IIFA using transfected HEK293 cells which express human AQP4 on their cell membranes. Results In total, 128 CNS IDD patients were studied. We found that 78% of NMO patients were seropositive for AQP4 autoantibodies by cell-based IIFA versus 61% by tissue-based IFA (p = 0.250, 75% of patients having relapsing myelitis (RM with LETM were seropositive by cell-based IIFA versus 50% by tissue-based IIFA (p = 0.250, and 33% of relapsing ON patients were seropositive by cell-based IIFA versus 22% by tissue-based IIFA (p = 1.000; however the differences were not statistically significant. All patients seropositive by tissue-based IIFA were also seropositive for AQP4 autoantibodies by cell-based IIFA. Among 29 NMOSD patients seropositive for AQP4 autoantibodies by cell-based IIFA, 20 (69% were seropositive by tissue-based IIFA. The 9 patients seropositive by cell-based IIFA while seronegative by tissue-based IIFA had NMO (3, RM with LETM (3, a single attack of LETM (1, relapsing ON (1 and a single ON attack (1. Among 23 NMO or RM patients seropositive for AQP4 autoantibodies by cell-based IIFA, comparison between those seropositive (n = 17 and seronegative (n = 6 by tissue-based IIFA revealed no differences in clinical and neuroradiological characteristics between the two groups. Conclusion Cell-based IIFA is slightly more sensitive

  8. A COMPARISON OF EASTERN AND WESTERN HONG KONG PHYTOPLANKTON FROM WEEKLY SAMPLES (1997-1999)

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Weekly phytoplankton samples were taken from western Hong Kong (Lamma) from Jan. 1997 to Dec. 1999 and from Port Shelter in Eastern Hong Kong from January 1998 to December 1999. During that time diatom blooms occurred repeatedly at both sites but never in synchrony. One species would bloom at one site and then weeks later it or another species would bloom at the other site; while the 1998 red tide of the mucus producing dinoflagellate Gymnodinium mikimotoi occurred at both sites. It first occurred at the Port Shelter site in March and did not appear at the Lamma site until April. With the single exception of this species, no other dinoflagellate reached bloom concentrations at the Lamma site. In addition, dinoflagellate abundance at the Lamma site was significantly lower (P <0.05) than that at the Port Shelter site. This was correlated with a significantly higher turbidity (i.e. low Secchi transparency) and higher turbulence (stronger currents) at the Lamma site.Annual variation in surface temperature correlated with total surface phytoplankton abundance at both our sample sites. Phytoplankton abundance increased in spring as water temperatures warmed. In fall, as surface water temperatures began to decline and the monsoon rains became less frequent there was a reduction in phytoplankton abundance associated with the reduction in temperature and light. Because so many variables co-occur with temperature (e.g. the amount of rainfall, light intensity and light duration etc.) it is not possible to cite temperature as the causal factor associated weth controlling phytoplankton abundance at our two sample sites.Our data support the rather controversial notion that percentage-wise, there are relatively more harmful bloom forming species in nutrient-rich coastal waters than there are in the world's oceans. 16% of the dinoflagellate species and 10.3% of the diatom species observed at our two sample sites were classed as harmful. These percentages were higher than those

  9. Characterization of coarse particulate matter in the western United States: a comparison between observation and modeling

    Directory of Open Access Journals (Sweden)

    R. Li

    2013-02-01

    Full Text Available We provide a regional characterization of coarse particulate matter (PM10–2.5 spanning the western United States based on the analysis of measurements from 50 sites reported in the US EPA Air Quality System (AQS and two state agencies. We found that the observed PM10–2.5 concentrations show significant spatial variability and distinct spatial patterns, associated with the distributions of land use/land cover and soil moisture. The highest concentrations were observed in the southwestern US, where sparse vegetation, shrublands or barren lands dominate with lower soil moistures, whereas the lowest concentrations were observed in areas dominated by grasslands, forest, or croplands with higher surface soil moistures. The observed PM10–2.5 concentrations also show variable seasonal, weekly, and diurnal patterns, indicating a variety of sources and their relative importance at different locations. The observed results were compared to modeled PM10–2.5 concentrations from an annual simulation using the Community Multiscale Air Quality modeling system (CMAQ that has been designed for regulatory or policy assessments of a variety of pollutants including PM10, which consists of PM10–2.5 and fine particulate matter (PM2.5. The model under-predicts PM10–2.5 observations at 49 of 50 sites, among which 14 sites have annual observation means that are at least five times greater than model means. Model results also fail to reproduce their spatial patterns. Important sources (e.g. pollen, bacteria, fungal spores, and geogenic dust were not included in the emission inventory used and/or the applied emissions were greatly under-estimated. Unlike the observed patterns that are more complex, modeled PM10–2.5 concentrations show the similar seasonal, weekly, and diurnal pattern; the temporal allocations in the modeling system need improvement. CMAQ does

  10. Characterization of coarse particulate matter in the western United States: a comparison between observation and modeling

    Science.gov (United States)

    Li, R.; Wiedinmyer, C.; Baker, K. R.; Hannigan, M. P.

    2013-02-01

    We provide a regional characterization of coarse particulate matter (PM10-2.5) spanning the western United States based on the analysis of measurements from 50 sites reported in the US EPA Air Quality System (AQS) and two state agencies. We found that the observed PM10-2.5 concentrations show significant spatial variability and distinct spatial patterns, associated with the distributions of land use/land cover and soil moisture. The highest concentrations were observed in the southwestern US, where sparse vegetation, shrublands or barren lands dominate with lower soil moistures, whereas the lowest concentrations were observed in areas dominated by grasslands, forest, or croplands with higher surface soil moistures. The observed PM10-2.5 concentrations also show variable seasonal, weekly, and diurnal patterns, indicating a variety of sources and their relative importance at different locations. The observed results were compared to modeled PM10-2.5 concentrations from an annual simulation using the Community Multiscale Air Quality modeling system (CMAQ) that has been designed for regulatory or policy assessments of a variety of pollutants including PM10, which consists of PM10-2.5 and fine particulate matter (PM2.5). The model under-predicts PM10-2.5 observations at 49 of 50 sites, among which 14 sites have annual observation means that are at least five times greater than model means. Model results also fail to reproduce their spatial patterns. Important sources (e.g. pollen, bacteria, fungal spores, and geogenic dust) were not included in the emission inventory used and/or the applied emissions were greatly under-estimated. Unlike the observed patterns that are more complex, modeled PM10-2.5 concentrations show the similar seasonal, weekly, and diurnal pattern; the temporal allocations in the modeling system need improvement. CMAQ does not include organic materials in PM10-2.5; however, speciation measurements show that organics constitute a significant component

  11. Comparison of colorimetric assays with quantitative amino acid analysis for protein quantification of Generalized Modules for Membrane Antigens (GMMA).

    Science.gov (United States)

    Rossi, Omar; Maggiore, Luana; Necchi, Francesca; Koeberling, Oliver; MacLennan, Calman A; Saul, Allan; Gerke, Christiane

    2015-01-01

    Genetically induced outer membrane particles from Gram-negative bacteria, called Generalized Modules for Membrane Antigens (GMMA), are being investigated as vaccines. Rapid methods are required for estimating the protein content for in-process assays during production. Since GMMA are complex biological structures containing lipid and polysaccharide as well as protein, protein determinations are not necessarily straightforward. We compared protein quantification by Bradford, Lowry, and Non-Interfering assays using bovine serum albumin (BSA) as standard with quantitative amino acid (AA) analysis, the most accurate currently available method for protein quantification. The Lowry assay has the lowest inter- and intra-assay variation and gives the best linearity between protein amount and absorbance. In all three assays, the color yield (optical density per mass of protein) of GMMA was markedly different from that of BSA with a ratio of approximately 4 for the Bradford assay, and highly variable between different GMMA; and approximately 0.7 for the Lowry and Non-Interfering assays, highlighting the need for calibrating the standard used in the colorimetric assay against GMMA quantified by AA analysis. In terms of a combination of ease, reproducibility, and proportionality of protein measurement, and comparability between samples, the Lowry assay was superior to Bradford and Non-Interfering assays for GMMA quantification.

  12. Analysis of ochratoxin A in grapes, musts and wines by LC–MS/MS: First comparison of stable isotope dilution assay and diastereomeric dilution assay methods

    Energy Technology Data Exchange (ETDEWEB)

    Roland, Aurélie, E-mail: aurelie@nyseos.fr [Nyseos, 2 place Pierre Viala, Montpellier Cedex 1 34060 (France); Bros, Pauline, E-mail: pauline.bros@gmail.com [Institut Français de la Vigne et du Vin, UMT Qualinnov, 2 place Pierre Viala, Montpellier Cedex 1 34060 (France); Bouisseau, Anaïs, E-mail: anais.bouisseau@gmail.com [Nyseos, 2 place Pierre Viala, Montpellier Cedex 1 34060 (France); Institut des Biomolécules Max Mousseron, UMR-CNRS-5247, Universités Montpellier I and II, Place Eugène Bataillon, 34095 Montpellier (France); Cavelier, Florine, E-mail: florine@univ-montp2.fr [Institut des Biomolécules Max Mousseron, UMR-CNRS-5247, Universités Montpellier I and II, Place Eugène Bataillon, 34095 Montpellier (France); Schneider, Rémi, E-mail: remi.schneider@vignevin.com [Institut Français de la Vigne et du Vin, UMT Qualinnov, 2 place Pierre Viala, Montpellier Cedex 1 34060 (France)

    2014-03-01

    Highlights: • OTA extraction on immunoaffinity columns is not adapted for DIDA quantification. • The use of a labeled internal standard is compulsory to obtain reliable results. • SIDA and DIDA quantification approaches have been compared for the first time. Abstract: Ochratoxin A (OTA) exhibits potent nephrotoxic, carcinogenic and teratogenic effects and its maximum level in wines has been set to 2 μg L⁻¹ by regulation. Consequently, the analytical procedures for OTA determination in wines have to be both very sensitive and reliable. In this paper, we compared two quantification methods: the stable isotope dilution assay (SIDA) and the diastereomeric dilution assay (DIDA). For this purpose, non-natural analogues of OTA were synthesized: the labeled OTA (OTA-d₄) as a diastereomeric mixture for the SIDA and one non-natural OTA’s diastereomer (OTA-dia) for the DIDA. To quantify OTA in red grapes, musts or wines, the sample preparation was optimized using immunoaffinity column extraction and the analysis was performed by LC–MS/MS in Multiple Reaction Monitoring mode. A validation procedure in agreement with the International Organization of Vine and Wine recommendations was conducted. It appeared that SIDA quantification exhibited excellent sensitivity (LOD < 1 ng L⁻¹), accuracy (recovery = 98%), repeatability (RSD < 3%) and intermediate reproducibility (RSD < 4%) compared to quantification by DIDA. Indeed, DIDA method did not provide satisfactory results demonstrating that immunoaffinity extraction is exclusively selective for the natural OTA and not for its diastereomer, which therefore cannot be considered as a good internal standard for this particular method.

  13. Investigation of watercourses by comparison of successive historical map surveys of Western Hungary

    Science.gov (United States)

    Kovács, Gábor

    2010-05-01

    The Second (Timár et al., 2006) and Third Military Survey (Biszak et al., 2007) of the Habsburg Empire, completed in the 19th century (1806-69 and 1869-87), can be very useful in different scientific investigations because of their accuracy and data content. The mapmakers used geodetic projections and survey technologies provided high accuracy. Therefore, scientists can use these maps and the represented objects in retrospective studies. The streams were drawn with very thin lines that also ascertain the high accuracy of their location. Previous study used the Second Military Survey to examine the neotectonic evaluation of the western part of the Pannonian Basin, bordered by Pinka, Rába and Répce Rivers (Kovács, 2010). The watercourses, especially alluvial ones, react very sensitively to tectonic forcing (Schumm & Khan, 1972; Ouchi, 1985). However, the present-day course of the creeks and rivers are mostly regulated, therefore they are unsuitable for such studies. The watercourses have reconstructed from maps surveyed prior to the main water control measures. The Second Military Survey was a perfect source for such studies. The investigated streams were free meandering ones. They could flood their banks, and only natural levees were present. After georeferencing the maps of the area, the streams were digitized, and their sinuosity values were computed. Where significant sinuosity changes have been detected along the streams, it can be considered as indicators of differential uplift or subsidence of the bedrock/alluvium. The goal of this study is to decide the character of several stream sections: were they free meandering ones or not? Some of the sections are antecedent ones, especially at the Vas Mountain at the present Austrian-Hungarian border. If the shapes of the watercourses on the different surveys are almost the same, the sinuosity refers to a prior, forced state of the stream. After digitizing the watercourses on the Third Military Survey sheets, some

  14. Tropospheric dry layers in the tropical western Pacific: comparisons of GPS radio occultation with multiple data sets

    Science.gov (United States)

    Rieckh, Therese; Anthes, Richard; Randel, William; Ho, Shu-Peng; Foelsche, Ulrich

    2017-03-01

    We use GPS radio occultation (RO) data to investigate the structure and temporal behavior of extremely dry, high-ozone tropospheric air in the tropical western Pacific during the 6-week period of the CONTRAST (CONvective TRansport of Active Species in the Tropics) experiment (January and February 2014). Our analyses are aimed at testing whether the RO method is capable of detecting these extremely dry layers and evaluating comparisons with in situ measurements, satellite observations, and model analyses. We use multiple data sources as comparisons, including CONTRAST research aircraft profiles, radiosonde profiles, AIRS (Atmospheric Infrared Sounder) satellite retrievals, and profiles extracted from the ERA (ERA-Interim reanalysis) and the GFS (US National Weather Service Global Forecast System) analyses, as well as MTSAT-2 satellite images. The independent and complementary radiosonde, aircraft, and RO data provide high vertical resolution observations of the dry layers. However, they all have limitations. The coverage of the radiosonde data is limited by having only a single station in this oceanic region; the aircraft data are limited in their temporal and spatial coverage; and the RO data are limited in their number and horizontal resolution over this period. However, nearby observations from the three types of data are highly consistent with each other and with the lower-vertical-resolution AIRS profiles. They are also consistent with the ERA and GFS data. We show that the RO data, used here for the first time to study this phenomenon, contribute significant information on the water vapor content and are capable of detecting layers in the tropics and subtropics with extremely low humidity (less than 10 %), independent of the retrieval used to extract moisture information. Our results also verify the quality of the ERA and GFS data sets, giving confidence to the reanalyses and their use in diagnosing the full four-dimensional structure of the dry layers.

  15. Determining antioxidant activities of lactobacilli cell-free supernatants by cellular antioxidant assay: a comparison with traditional methods.

    Directory of Open Access Journals (Sweden)

    Jiali Xing

    Full Text Available Antioxidant activity of lactic acid bacteria is associated with multiple health-protective effects. Traditional indexes of chemical antioxidant activities poorly reflect the antioxidant effects of these bacteria in vivo. Cellular antioxidant activity (CAA assay was used in this study to determine the antioxidant activity of cell-free supernatants (CFSs of 10 Lactobacillus strains. The performance of the CAA assay was compared with that of four chemical antioxidant activity assays, namely, DPPH radical scavenging, hydroxyl radical scavenging (HRS, reducing power (RP, and inhibition of linoleic acid peroxidation (ILAP. Results of the CAA assay were associated with those of DPPH and ILAP assays, but not with those of RP and HRS assays. The inter- and intra-specific antioxidant activities of CFS were characterized by chemical and CAA assays. L. rhamnosus CCFM 1107 displayed a high antioxidative effect similar to positive control L. rhamnosus GG ATCC 53103 in all of the assays. The CAA assay is a potential method for the detection of antioxidant activities of lactobacilli CFSs.

  16. Comparison of the ELISPOT and cytokine flow cytometry assays for the enumeration of antigen-specific T cells.

    Science.gov (United States)

    Karlsson, Annika C; Martin, Jeffrey N; Younger, Sophie R; Bredt, Barry M; Epling, Lorrie; Ronquillo, Rollie; Varma, Arjun; Deeks, Steven G; McCune, Joseph M; Nixon, Douglas F; Sinclair, Elizabeth

    2003-12-01

    The enumeration of antigen-specific T cell responses has been greatly facilitated in recent years by the development of methods based on the detection of cytokines. In particular, the enzyme-linked immunospot (ELISPOT) and cytokine flow cytometry (CFC) assays have become popular. Since both assays are likely to continue to be in widespread use, it is important to evaluate whether their results are comparable. In the current study, we compared the results obtained in the ELISPOT and CFC assays using peptide pools corresponding to CMV and HIV-1 proteins in chronically HIV-1-infected individuals. Analysis of T cell responses to peptide pools indicated that the CMV pp65 and HIV-1 Gag CFC and ELISPOT-derived results were statistically correlated. However, the results obtained with each assay differed in important ways: the magnitude of the response was consistently higher in the CFC assay while the CFC assay was less likely than the ELISPOT assay to detect low-level responses. Furthermore, there was a lack of numeric agreement between ELISPOT and CFC results. For studies that require the detection of low-level responses, or definition of responses as positive or negative, the ELISPOT assay may be preferable. In contrast, the CFC has a greater dynamic range and allows for phenotypic discrimination of responding cells, making it the assay of choice for most other applications.

  17. Prognostic Ability of Practitioners of Traditional Arabic Medicine: Comparison with Western Methods through a Relative Patient Progress Scale

    Directory of Open Access Journals (Sweden)

    Bertrand Graz

    2010-01-01

    Full Text Available The ancient Greek medical theory based on balance or imbalance of humors disappeared in the western world, but does survive elsewhere. Is this survival related to a certain degree of health care efficiency? We explored this hypothesis through a study of classical Greco-Arab medicine in Mauritania. Modern general practitioners evaluated the safety and effectiveness of classical Arabic medicine in a Mauritanian traditional clinic, with a prognosis/follow-up method allowing the following comparisons: (i actual patient progress (clinical outcome compared with what the traditional ‘tabib’ had anticipated (= prognostic ability and (ii patient progress compared with what could be hoped for if the patient were treated by a modern physician in the same neighborhood. The practice appeared fairly safe and, on average, clinical outcome was similar to what could be expected with modern medicine. In some cases, patient progress was better than expected. The ability to correctly predict an individual's clinical outcome did not seem to be better along modern or Greco-Arab theories. Weekly joint meetings (modern and traditional practitioners were spontaneously organized with a modern health centre in the neighborhood. Practitioners of a different medical system can predict patient progress. For the patient, avoiding false expectations with health care and ensuring appropriate referral may be the most important. Prognosis and outcome studies such as the one presented here may help to develop institutions where patients find support in making their choices, not only among several treatment options, but also among several medical systems.

  18. Comparison of Droplet Digital PCR and Quantitative PCR Assays for Quantitative Detection of Xanthomonas citri Subsp. citri.

    Directory of Open Access Journals (Sweden)

    Yun Zhao

    Full Text Available Droplet digital polymerase chain reaction (ddPCR is a novel molecular biology technique providing absolute quantification of target nucleic acids without the need for an external calibrator. Despite its emerging applications in medical diagnosis, there are few reports of its use for the detection of plant pathogens. This work was designed to assess the diagnosis potential of the ddPCR for absolute quantitative detection of Xanthomonas citri subsp. citri, a quarantine plant pathogenic bacterium that causes citrus bacterial canker in susceptible Citrus species. We transferred an established quantitative PCR (qPCR assay for citrus bacterial canker diagnosis directly to the ddPCR format and compared the performance of the two methods. The qPCR assay has a broader dynamic range compared to the ddPCR assay and the ddPCR assay has a significantly higher degree of sensitivity compared to the qPCR assay. The influence of PCR inhibitors can be reduced considerably in the ddPCR assay because the collection of end-point fluorescent signals and the counting of binomial events (positive or negative droplets are associated with a Poisson algorithm. The ddPCR assay also shows lower coefficient of variation compared to the qPCR assay especially in low target concentration. The linear association of the measurements by ddPCR and qPCR assays is strong (Pearson correlation = 0.8633; P<0.001. Receiver operating characteristic analysis indicates the ddPCR methodology is a more robust approach for diagnosis of citrus bacterial canker. In summary, the results demonstrated that the ddPCR assay has the potential for the quantitative detection of X. citri subsp. citri with high precision and accuracy as compared with the results from qPCR assay. Further studies are required to evaluate and validate the value of ddPCR technology in the diagnosis of plant disease and quarantine applications.

  19. Comparison of two real-time RT-PCR assays for differentiation of C-strain vaccinated from classical swine fever infected pigs and wild boars.

    Science.gov (United States)

    Widén, F; Everett, H; Blome, S; Fernandez Pinero, J; Uttenthal, A; Cortey, M; von Rosen, T; Tignon, M; Liu, L

    2014-10-01

    Classical swine fever is one of the most important infectious diseases for the pig industry worldwide due to its economic impact. Vaccination is an effective means to control disease, however within the EU its regular use is banned owing to the inability to differentiate infected and vaccinated animals, the so called DIVA principle. This inability complicates monitoring of disease and stops international trade thereby limiting use of the vaccine in many regions. The C-strain vaccine is safe to use and gives good protection. It is licensed for emergency vaccination in the EU in event of an outbreak. Two genetic assays that can distinguish between wild type virus and C-strain vaccines have recently been developed. Here the results from a comparison of these two real-time RT-PCR assays in an interlaboratory exercise are presented. Both assays showed similar performance.

  20. Comparison of Intact PTH and Bio-Intact PTH Assays Among Non-Dialysis Dependent Chronic Kidney Disease Patients.

    Science.gov (United States)

    Einbinder, Yael; Benchetrit, Sydney; Golan, Eliezer; Zitman-Gal, Tali

    2017-09-01

    The third-generation bio-intact parathyroid hormone (PTH) (1-84) assay was designed to overcome problems associated with the detection of C-terminal fragments by the second-generation intact PTH assay. The two assays have been compared primarily among dialysis populations. The present study evaluated the correlations and differences between these two PTH assays among patients with chronic kidney disease (CKD) stages 3 to 5 not yet on dialysis. Blood samples were collected from 98 patients with CKD stages 3 to 5. PTH concentrations were measured simultaneously by using the second-generation - PTH intact-STAT and third-generation bio-intact 1-84 PTH assays. Other serum biomarkers of bone mineral disorders were also assessed. CKD stage was calculated by using the CKD-Epidemiology Collaboration (EPI) formula. Serum bio-intact PTH concentrations were strongly correlated but significantly lower than the intact PTH concentrations (r=0.963, Pbio-intact PTH) positively correlated with urea (r=0.523, r=0.504; P=0.002, respectively), phosphorus (r=0.532, r=0.521; Pbio-intact PTH assay detected significantly lower PTH concentrations compared with intact PTH assay. Additional studies that correlate the diagnosis and management of CKD mineral and bone disorders with bone histomorphometric findings are needed to determine whether bio-intact PTH assay results are better surrogate markers in these early stages of CKD.

  1. Bench-top validation testing of selected immunological and molecular Renibacterium salmoninarum diagnostic assays by comparison with quantitative bacteriological culture

    Science.gov (United States)

    Elliott, D.G.; Applegate, L.J.; Murray, A.L.; Purcell, M.K.; McKibben, C.L.

    2013-01-01

    No gold standard assay exhibiting error-free classification of results has been identified for detection of Renibacterium salmoninarum, the causative agent of salmonid bacterial kidney disease. Validation of diagnostic assays for R. salmoninarum has been hindered by its unique characteristics and biology, and difficulties in locating suitable populations of reference test animals. Infection status of fish in test populations is often unknown, and it is commonly assumed that the assay yielding the most positive results has the highest diagnostic accuracy, without consideration of misclassification of results. In this research, quantification of R. salmoninarum in samples by bacteriological culture provided a standardized measure of viable bacteria to evaluate analytical performance characteristics (sensitivity, specificity and repeatability) of non-culture assays in three matrices (phosphate-buffered saline, ovarian fluid and kidney tissue). Non-culture assays included polyclonal enzyme-linked immunosorbent assay (ELISA), direct smear fluorescent antibody technique (FAT), membrane-filtration FAT, nested polymerase chain reaction (nested PCR) and three real-time quantitative PCR assays. Injection challenge of specific pathogen-free Chinook salmon, Oncorhynchus tshawytscha (Walbaum), with R. salmoninarum was used to estimate diagnostic sensitivity and specificity. Results did not identify a single assay demonstrating the highest analytical and diagnostic performance characteristics, but revealed strengths and weaknesses of each test.

  2. A prospective comparison of molecular assay and touch imprint cytology for intraoperative evaluation of sentinel lymph nodes

    Institute of Scientific and Technical Information of China (English)

    CHEN Jia-jian; SHEN Zhen-zhou; WANG Yong-sheng; WU Jiong; YANG Ben-long; CHEN Jia-ying; ZHANG Jia-xin; LI Da-li; XU Wei-ping; XU Xiao-li; YANG Wen-tao; SHAO Zhi-min

    2011-01-01

    Background Accurate intraoperative diagnosis of sentinel lymph node (SLN) metastases enables the selection of patients for axillary lymph node dissections during the same operation, reducing the need for a second operation. The present study aimed to prospectively compare the GeneSearchTM Breast Lymph Node (BLN) Assay with touch imprint cytology (TIC) for intraoperative evaluation of SLNs.Methods SLNs were sectioned in 1.5-3.0 mm pieces. TIC was performed on all pieces and the BLN Assay and postoperative histology evaluations were performed on different alternating node pieces. Overall performance of the BLN Assay was compared with that of TIC relative to the postoperative histology results.Results A total of 90 patients enrolled in the study. Complete intraoperative data for both the BLN Assay and TIC were collected in 86 patients. The sensitivity, specificity, and overall accuracy of the BLN Assay were 82%, 97%, and 92%,respectively on a per patient basis compared with those of TIC which were 67%, 100%, and 90%.Conclusions Performance of the BLN Assay was superior to that of TIC and the additional application of TIC did not help improve the total sensitivity and accuracy of the intraoperative assessment. The existence of ectopic breast tissue might be a possible cause of false positive for the BLN assay. In addition, the BLN Assay complements histopathology assessment and can minimize sampling error without increasing pathologists' workload.

  3. Bench-top validation testing of selected immunological and molecular Renibacterium salmoninarum diagnostic assays by comparison with quantitative bacteriological culture.

    Science.gov (United States)

    Elliott, D G; Applegate, L J; Murray, A L; Purcell, M K; McKibben, C L

    2013-09-01

    No gold standard assay exhibiting error-free classification of results has been identified for detection of Renibacterium salmoninarum, the causative agent of salmonid bacterial kidney disease. Validation of diagnostic assays for R. salmoninarum has been hindered by its unique characteristics and biology, and difficulties in locating suitable populations of reference test animals. Infection status of fish in test populations is often unknown, and it is commonly assumed that the assay yielding the most positive results has the highest diagnostic accuracy, without consideration of misclassification of results. In this research, quantification of R. salmoninarum in samples by bacteriological culture provided a standardized measure of viable bacteria to evaluate analytical performance characteristics (sensitivity, specificity and repeatability) of non-culture assays in three matrices (phosphate-buffered saline, ovarian fluid and kidney tissue). Non-culture assays included polyclonal enzyme-linked immunosorbent assay (ELISA), direct smear fluorescent antibody technique (FAT), membrane-filtration FAT, nested polymerase chain reaction (nested PCR) and three real-time quantitative PCR assays. Injection challenge of specific pathogen-free Chinook salmon, Oncorhynchus tshawytscha (Walbaum), with R. salmoninarum was used to estimate diagnostic sensitivity and specificity. Results did not identify a single assay demonstrating the highest analytical and diagnostic performance characteristics, but revealed strengths and weaknesses of each test.

  4. Comparison of in vivo and in vitro reporter gene assays for short-term screening of estrogenic activity

    NARCIS (Netherlands)

    Legler, J.; Zeinstra, L.M.; Schuitemaker, F.; Lanser, P.H.; Bogerd, J.; Brouwer, A.; Vethaak, A.D.; Voogt, de P.; Murk, A.J.; Burg, van der B.

    2002-01-01

    Functional in vitro and in vivo reporter gene assays have recently been developed for the rapid determination of exposure to (xeno)estrogens. The in vitro estrogen receptor (ER)-mediated chemically activated luciferase gene expression (ER-CALUX) assay uses T47D human breast cancer cells stably trans

  5. Comparison of the latex agglutination test with the hemagglutination inhibition test, enzyme-linked immunosorbent assay, and neutralization test for detection of antibodies to rubella virus.

    OpenAIRE

    Meegan, J M; Evans, B. K.; Horstmann, D. M.

    1982-01-01

    The ability of a rapid, latex agglutination test to diagnose rubella infection and to measure immune status was evaluated by comparison with the hemagglutination-inhibition (HAI) test, enzyme-linked immunosorbent assay (ELISA), and the neutralization (NT) test. The latex agglutination test accurately detected serological conversions in 74 pairs of sera representing 21 natural infections and 53 immunizations. The antibody levels of 276 sera from the general population were determined by latex ...

  6. Comparison of two real-time PCR assays for the detection of malaria parasites from hemolytic blood samples - Short communication.

    Science.gov (United States)

    Hagen, Ralf Matthias; Hinz, Rebecca; Tannich, Egbert; Frickmann, Hagen

    2015-06-01

    We compared the performance of an in-house and a commercial malaria polymerase chain reaction (PCR) assay using freeze-thawed hemolytic blood samples. A total of 116 freeze-thawed ethylenediamine tetraacetic acid (EDTA) blood samples of patients with suspicion of malaria were analyzed by an in-house as well as by a commercially available real-time PCR. Concordant malaria negative PCR results were reported for 39 samples and malaria-positive PCR results for 67 samples. The in-house assay further detected one case of Plasmodium falciparum infection, which was negative in the commercial assay as well as five cases of P. falciparum malaria and three cases of Plasmodium vivax malaria, which showed sample inhibition in the commercial assay. The commercial malaria assay was positive in spite of a negative in-house PCR result in one case. In all concordant results, cycle threshold values of P. falciparum-positive samples were lower in the commercial PCR than in the in-house assay. Although Ct values of the commercial PCR kit suggest higher sensitivity in case of concordant results, it is prone to inhibition if it is applied to hemolytic freeze-thawed blood samples. The number of misidentifications was, however, identical for both real-time PCR assays.

  7. PCR assay detects Mannheimia haemolytica in culture-negative pneumonic lung tissues of bighorn sheep (Ovis canadensis) from outbreaks in the western USA, 2009-2010.

    Science.gov (United States)

    Shanthalingam, Sudarvili; Goldy, Andrea; Bavananthasivam, Jegarubee; Subramaniam, Renuka; Batra, Sai Arun; Kugadas, Abirami; Raghavan, Bindu; Dassanayake, Rohana P; Jennings-Gaines, Jessica E; Killion, Halcyon J; Edwards, William H; Ramsey, Jennifer M; Anderson, Neil J; Wolff, Peregrine L; Mansfield, Kristin; Bruning, Darren; Srikumaran, Subramaniam

    2014-01-01

    Mannheimia haemolytica consistently causes severe bronchopneumonia and rapid death of bighorn sheep (Ovis canadensis) under experimental conditions. However, Bibersteinia trehalosi and Pasteurella multocida have been isolated from pneumonic bighorn lung tissues more frequently than M. haemolytica by culture-based methods. We hypothesized that assays more sensitive than culture would detect M. haemolytica in pneumonic lung tissues more accurately. Therefore, our first objective was to develop a PCR assay specific for M. haemolytica and use it to determine if this organism was present in the pneumonic lungs of bighorns during the 2009-2010 outbreaks in Montana, Nevada, and Washington, USA. Mannheimia haemolytica was detected by the species-specific PCR assay in 77% of archived pneumonic lung tissues that were negative by culture. Leukotoxin-negative M. haemolytica does not cause fatal pneumonia in bighorns. Therefore, our second objective was to determine if the leukotoxin gene was also present in the lung tissues as a means of determining the leukotoxicity of M. haemolytica that were present in the lungs. The leukotoxin-specific PCR assay detected leukotoxin gene in 91% of lung tissues that were negative for M. haemolytica by culture. Mycoplasma ovipneumoniae, an organism associated with bighorn pneumonia, was detected in 65% of pneumonic bighorn lung tissues by PCR or culture. A PCR assessment of distribution of these pathogens in the nasopharynx of healthy bighorns from populations that did not experience an all-age die-off in the past 20 yr revealed that M. ovipneumoniae was present in 31% of the animals whereas leukotoxin-positive M. haemolytica was present in only 4%. Taken together, these results indicate that culture-based methods are not reliable for detection of M. haemolytica and that leukotoxin-positive M. haemolytica was a predominant etiologic agent of the pneumonia outbreaks of 2009-2010.

  8. Seroprevalence of anti-hepatitis E virus (HEV in a Korean population: comparison of two commercial anti-HEV assays

    Directory of Open Access Journals (Sweden)

    Park Hyun

    2012-06-01

    Full Text Available Abstract Background Hepatitis E virus (HEV has emerged as an important cause of epidemic and sporadic acute viral hepatitis worldwide. This study investigated the seroprevalence of anti-HEV in a Korean population and compared the performance of two commercially available anti-HEV assays. Methods A total 147 health-check examinees were randomly sampled as matched to the age- and sex- adjusted standard population based on the Korean National Census of 2007. Serum immunoglobulin G anti-HEV was determined by using the Genelabs assay (Genelabs, Singapore and the Wantai assay (Wantai, Beijing, China. Results The overall anti-HEV seroprevalence was 23.1% (95% CI, 16.1-30.1% using the Wantai assay and 14.3% (95% CI, 8.3-20.3% using the Genelabs assay. Only 12 samples (8.1% were positive for anti-HEV as measured by both assays; agreement between the two assays was poor (kappa value of 0.315. The anti-HEV seroprevalence increased with age from 2% and 3% in the people younger than 20-years-of-age to 34.6% and 42.3% in those over 59-years-of-age by the Genelabs and Wantai assay, respectively. Conclusions The HEV seroprevalence in Korean population is about 20% overall, with seroprevalence increasing in this population with increasing age. There was poor concordance in the results of the Genelabs and Wantai assays, which warrants further study concerning a reliable diagnostic test for the diagnosis of hepatitis E.

  9. HPV16 seropositivity and subsequent HPV16 infection risk in a naturally infected population: comparison of serological assays.

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    Shih-Wen Lin

    Full Text Available BACKGROUND: Several serological assays have been developed to detect antibodies elicited against infections with oncogenic human papillomavirus (HPV type 16. The association between antibody levels measured by various assays and subsequent HPV infection risk may differ. We compared HPV16-specific antibody levels previously measured by a virus-like particle (VLP-based direct enzyme-linked immunoassay (ELISA with levels measured by additional assays and evaluated the protection against HPV16 infection conferred at different levels of the assays. METHODOLOGY/PRINCIPAL FINDINGS: Replicate enrollment serum aliquots from 388 unvaccinated women in the control arm of the Costa Rica HPV vaccine trial were measured for HPV16 seropositivity using three serological assays: a VLP-based direct ELISA; a VLP-based competitive Luminex immunoassay (cLIA; and a secreted alkaline phosphatase protein neutralization assay (SEAP-NA. We assessed the association of assay seropositivity and risk of subsequent HPV16 infection over four years of follow-up by calculating sampling-adjusted odds ratios (OR and HPV16 seropositivity based on standard cutoff from the cLIA was significantly associated with protection from subsequent HPV16 infection (OR = 0.48, CI = 0.27-0.86, compared with seronegatives. Compared with seronegatives, the highest seropositive tertile antibody levels from the direct ELISA (OR = 0.53, CI = 0.28-0.90 as well as the SEAP-NA (OR = 0.20, CI = 0.06, 0.64 were also significantly associated with protection from HPV16 infection. CONCLUSIONS/SIGNIFICANCE: Enrollment HPV16 seropositivity by any of the three serological assays evaluated was associated with protection from subsequent infection, although cutoffs for immune protection were different. We defined the assays and seropositivity levels after natural infection that better measure and translate to protective immunity.

  10. A comparison of two measures of HIV diversity in multi-assay algorithms for HIV incidence estimation.

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    Matthew M Cousins

    Full Text Available Multi-assay algorithms (MAAs can be used to estimate HIV incidence in cross-sectional surveys. We compared the performance of two MAAs that use HIV diversity as one of four biomarkers for analysis of HIV incidence.Both MAAs included two serologic assays (LAg-Avidity assay and BioRad-Avidity assay, HIV viral load, and an HIV diversity assay. HIV diversity was quantified using either a high resolution melting (HRM diversity assay that does not require HIV sequencing (HRM score for a 239 base pair env region or sequence ambiguity (the percentage of ambiguous bases in a 1,302 base pair pol region. Samples were classified as MAA positive (likely from individuals with recent HIV infection if they met the criteria for all of the assays in the MAA. The following performance characteristics were assessed: (1 the proportion of samples classified as MAA positive as a function of duration of infection, (2 the mean window period, (3 the shadow (the time period before sample collection that is being assessed by the MAA, and (4 the accuracy of cross-sectional incidence estimates for three cohort studies.The proportion of samples classified as MAA positive as a function of duration of infection was nearly identical for the two MAAs. The mean window period was 141 days for the HRM-based MAA and 131 days for the sequence ambiguity-based MAA. The shadows for both MAAs were <1 year. Both MAAs provided cross-sectional HIV incidence estimates that were very similar to longitudinal incidence estimates based on HIV seroconversion.MAAs that include the LAg-Avidity assay, the BioRad-Avidity assay, HIV viral load, and HIV diversity can provide accurate HIV incidence estimates. Sequence ambiguity measures obtained using a commercially-available HIV genotyping system can be used as an alternative to HRM scores in MAAs for cross-sectional HIV incidence estimation.

  11. Study of a viral-dual infection in rainbow trout (Oncorhynchus mykiss) by seroneutralization, western blot and polymerase chain reaction assays.

    Science.gov (United States)

    Rodríguez, S; Vilas, M P; Alonso, M; Pérez, S I

    1995-12-01

    Viral-dual infections in fish are of interest to aquaculture practices but they are rarely described and studied. Several methods were applied in this work to demonstrate a case of coinfection in a reared rainbow trout (Oncorhynchus mykiss) population. Inoculation in cell cultures and cross-neutralization tests were the standard procedures that made it possible to isolate and identify a birnavirus, the infectious pancreatic necrosis virus (IPNV), and suspect of a second virus. Western blotting with both polyclonal and monoclonal antibodies, and reverse transcriptional-polymerase chain reaction (RT-PCR) demonstrate coexistence of both, IPNV and a rhabdovirus.

  12. Comparison of in vitro eye irritation potential by bovine corneal opacity and permeability (BCOP) assay to erythema scores in human eye sting test of surfactant-based formulations.

    Science.gov (United States)

    Cater, Kathleen C; Harbell, John W

    2008-01-01

    The bovine corneal opacity and permeability (BCOP) assay can be used to predict relative eye irritation potential of surfactant-based personal care formulations relative to a corporate benchmark. The human eye sting test is typically used to evaluate product claims of no tears/no stinging for children's bath products. A preliminary investigation was conducted to test a hypothesis that the BCOP assay could be used as a prediction model for relative ranking of human eye irritation responses under conditions of a standard human eye sting test to surfactant-based formulations. BCOP assays and human eye sting tests were conducted on 4 commercial and 1 prototype body wash (BW) developed specifically for children or as mild bath products. In the human eye sting test, 10 mul of a 10% dosing solution is instilled into one eye of each panelist (n = 20), and the contralateral eye is dosed with sterile water as a control. Bulbar conjunctival erythema responses of each eye are graded at 30 seconds by an ophthalmologist. The BCOP assay permeability values (optical density at 490 nm [OD(490)]) for the 5 BWs ranged from 0.438 to 1.252 (i.e., least to most irritating). By comparison, the number of panelists exhibiting erythema responses (mild to moderately pink) ranged from 3 of 20 panelists for the least irritating BW to 10 of 20 panelists for the most irritating BW tested. The relative ranking of eye irritation potential of the 5 BWs in the BCOP assay compares favorably with the relative ranking of the BWs in the human eye sting test. Based on these findings, the permeability endpoint of the BCOP assay, as described for surfactant-based formulations, showed promise as a prediction model for relative ranking of conjunctival erythema responses in the human eye. Consequently, screening of prototype formulations in the BCOP assay would allow for formula optimization of mild bath products prior to investment in a human eye sting test.

  13. Airborne measurements of western U.S. wildfire emissions: Comparison with prescribed burning and air quality implications: Western U.S. Wildfire Emissions

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Xiaoxi [School of Earth and Atmospheric Sciences, Georgia Institute of Technology, Atlanta Georgia USA; Now at Cooperative Institute for Research in Environmental Sciences, University of Colorado Boulder, Boulder Colorado USA; Now at Department of Chemistry, University of Colorado Boulder, Boulder Colorado USA; Huey, L. Gregory [School of Earth and Atmospheric Sciences, Georgia Institute of Technology, Atlanta Georgia USA; Yokelson, Robert J. [Department of Chemistry, University of Montana, Missoula Montana USA; Selimovic, Vanessa [Department of Chemistry, University of Montana, Missoula Montana USA; Simpson, Isobel J. [Department of Chemistry, University of California, Irvine California USA; Müller, Markus [Department of Chemistry, University of Montana, Missoula Montana USA; Institute for Ion Physics and Applied Physics, University of Innsbruck, Innsbruck Austria; Jimenez, Jose L. [Cooperative Institute for Research in Environmental Sciences, University of Colorado Boulder, Boulder Colorado USA; Department of Chemistry, University of Colorado Boulder, Boulder Colorado USA; Campuzano-Jost, Pedro [Cooperative Institute for Research in Environmental Sciences, University of Colorado Boulder, Boulder Colorado USA; Department of Chemistry, University of Colorado Boulder, Boulder Colorado USA; Beyersdorf, Andreas J. [NASA Langley Research Center, Hampton Virginia USA; Now at Department of Chemistry, California State University, San Bernardino California USA; Blake, Donald R. [Department of Chemistry, University of California, Irvine California USA; Butterfield, Zachary [Earth and Environmental Sciences Division, Los Alamos National Laboratory, Los Alamos New Mexico USA; Now at Department of Atmospheric, Oceanic, and Space Sciences, University of Michigan, Ann Arbor Michigan USA; Choi, Yonghoon [NASA Langley Research Center, Hampton Virginia USA; Science Systems and Applications, Inc., Hampton Virginia USA; Crounse, John D. [Division of Geological and Planetary Sciences, California Institute of Technology, Pasadena California USA; Day, Douglas A. [Cooperative Institute for Research in Environmental Sciences, University of Colorado Boulder, Boulder Colorado USA; Department of Chemistry, University of Colorado Boulder, Boulder Colorado USA; Diskin, Glenn S. [NASA Langley Research Center, Hampton Virginia USA; Dubey, Manvendra K. [Earth and Environmental Sciences Division, Los Alamos National Laboratory, Los Alamos New Mexico USA; Fortner, Edward [Center for Aerosol and Cloud Chemistry, Aerodyne Research Inc., Billerica Massachusetts USA; Hanisco, Thomas F. [Atmospheric Chemistry and Dynamics Laboratory, NASA Goddard Space Flight Center, Greenbelt Maryland USA; Hu, Weiwei [Cooperative Institute for Research in Environmental Sciences, University of Colorado Boulder, Boulder Colorado USA; Department of Chemistry, University of Colorado Boulder, Boulder Colorado USA; King, Laura E. [School of Earth and Atmospheric Sciences, Georgia Institute of Technology, Atlanta Georgia USA; Kleinman, Lawrence [Environmental and Climate Sciences Department, Brookhaven National Laboratory, Upton New York USA; Meinardi, Simone [Department of Chemistry, University of California, Irvine California USA; Mikoviny, Tomas [Department of Chemistry, University of Oslo, Oslo Norway; Onasch, Timothy B. [Center for Aerosol and Cloud Chemistry, Aerodyne Research Inc., Billerica Massachusetts USA; Palm, Brett B. [Cooperative Institute for Research in Environmental Sciences, University of Colorado Boulder, Boulder Colorado USA; Department of Chemistry, University of Colorado Boulder, Boulder Colorado USA; Peischl, Jeff [Cooperative Institute for Research in Environmental Sciences, University of Colorado Boulder, Boulder Colorado USA; Earth System Research Laboratory, National Oceanic and Atmospheric Administration, Boulder Colorado USA; Pollack, Ilana B. [Cooperative Institute for Research in Environmental Sciences, University of Colorado Boulder, Boulder Colorado USA; Earth System Research Laboratory, National Oceanic and Atmospheric Administration, Boulder Colorado USA

    2017-06-14

    Wildfires emit significant amounts of pollutants that degrade air quality. Plumes from three wildfires in the western U.S. were measured from aircraft during the Studies of Emissions and Atmospheric Composition, Clouds, and Climate Coupling by Regional Surveys (SEAC4RS) and the Biomass Burning Observation Project (BBOP), both in summer 2013. This study reports an extensive set of emission factors (EFs) for over 80 gases and 5 components of submicron particulate matter (PM1) from these temperate wildfires. These include rarely, or never before, measured oxygenated volatile organic compounds and multifunctional organic nitrates. The observed EFs are compared with previous measurements of temperate wildfires, boreal forest fires, and temperate prescribed fires. The wildfires emitted high amounts of PM1 (with organic aerosol (OA) dominating the mass) with an average EF that is more than two times the EFs for prescribed fires. The measured EFs were used to estimate the annual wildfire emissions of carbon monoxide, nitrogen oxides, total nonmethane organic compounds, and PM1 from 11 western U.S. states. The estimated gas emissions are generally comparable with the 2011 National Emissions Inventory (NEI). However, our PM1 emission estimate (1530 ± 570 Gg yr-1) is over three times that of the NEI PM2.5 estimate and is also higher than the PM2.5 emitted from all other sources in these states in the NEI. This study indicates that the source of OA from BB in the western states is significantly underestimated. In addition, our results indicate prescribed burning may be an effective method to reduce fine particle emissions.

  14. Comparison of in vivo and in vitro reporter gene assays for short-term screening of estrogenic activity.

    Science.gov (United States)

    Legler, Juliette; Zeinstra, Laura M; Schuitemaker, Femke; Lanser, Peter H; Bogerd, Jan; Brouwer, Abraham; Vethaak, A Dick; De Voogt, Pim; Murk, Albertinka J; Van der Burg, Bart

    2002-10-15

    Functional in vitro and in vivo reporter gene assays have recently been developed for the rapid determination of exposure to (xeno)estrogens. The in vitro estrogen receptor (ER)-mediated chemically activated luciferase gene expression (ER-CALUX) assay uses T47D human breast cancer cells stably transfected with an ER-mediated luciferase gene construct. In the in vivo assay, transgenic zebrafish are used in which the same luciferase construct has been stably introduced. In both assays, luciferase reporter gene activity can be easily quantified following short-term exposure to chemicals activating endogenous estrogen receptors. The objective of this study was to compare responses by known (xeno)estrogenic compounds in both assays. Exposure to the (xeno)estrogens estradiol (E2), estrone, ethynylestradiol (EE2), o,p'-DDT, nonylphenol (NP), and di(2-ethylhexyl)phthalate (DEHP) revealed that EE2 was the most potent (xeno)estrogen tested and was 100 times more potent than E2 in the transgenic zebrafish assay, whereas in the in vitro ER-CALUX assay, EE2 and E2 were equipotent Although the xenoestrogens o,p'-DDT and NP were full estrogen agonists in the in vitro ER-CALUX assay, only o,p'-DDT demonstrated weak dose-related estrogenic activity in vivo. To determine if differences in reporter gene activity may be explained by differential affinity of (xeno)estrogens to human and zebrafish ERs, full-length sequences of the zebrafish ER subtypes alpha, beta, and gamma were cloned, and transactivation by (xeno)estrogens was compared to human ERalpha and ERbeta. Using transiently transfected recombinant ER and reporter gene constructs, EE2 also showed relatively potent activation of zebrafish ERalpha and ERbeta compared to human ERalpha and ERbeta. Zebrafish ERbeta and ERgamma showed higher transactivation by (xeno)estrogens relative to E2 than human ERbeta.

  15. Comparison of three different commercial PCR assays for the detection of pathogens in critically ill sepsis patients.

    Science.gov (United States)

    Schreiber, J; Nierhaus, A; Braune, S A; de Heer, G; Kluge, S

    2013-05-01

    The high mortality rate associated with sepsis necessitates a timely identification of the causative organism in order to optimize antimicrobial therapy. PCR assays are increasingly being used for this purpose. The aim of this study was to compare three commercially available PCR systems for the diagnosis of systemic infections. In a prospective observational study, a broad-range (SepsiTest®; Molzym, Bremen, Germany) and two multiplex PCR assays (VYOO®; SIRS-Lab, Jena, Germany and LightCycler® SeptiFast; Roche, Mannheim, Germany) were compared to blood cultures with respect to the clinical course of 50 critically ill patients with sepsis, severe sepsis or septic shock. Pathogens were detected by PCR in 12 % (SepsiTest®), 10 % (VYOO®) and 14 % (LightCycler® SeptiFast) of samples and in 26 % by blood culture. Negative results were obtained using all four methods in 32 samples (64 %) and 3 (6 %) samples were positive in all tests. Upon consideration of additional diagnostic findings and the clinical course, eight (16 %) of the positive blood culture results were deemed clinically relevant. All three PCR assays could also identify the causative organism (or a specific gene thereof) in three of these eight positive blood cultures, whereas for five of the eight, all three PCR assays were negative. In one patient with a negative blood culture, the SepsiTest®, VYOO® and LightCycler® SeptiFast assays were positive for Streptococcus species. The PCR assays appeared to be less susceptible than blood cultures to false-positive results arising from contamination with coagulase-negative staphylococcal organisms. There was some variability between the three PCR assays tested and the corresponding blood cultures with regards to the type of pathogen detected. The three PCR assays appeared to be less susceptible to false-positive results than blood cultures.

  16. Results from an inter-laboratory comparison of pneumococcal serotype-specific IgG measurement and critical parameters that affect assay performance.

    Science.gov (United States)

    Balloch, A; Licciardi, P V; Leach, A; Nurkka, A; Tang, M L K

    2010-02-01

    Quantitation of specific IgG to polysaccharides (serotypes) of Streptococcus pneumoniae provides the basis for evaluating vaccine efficacy. Different enzyme-linked immunosorbent assay (ELISA) methods are used internationally, making comparisons between laboratories difficult. We undertook an inter-laboratory comparison between two international laboratories performing serotype-specific IgG ELISAs using a panel of well-characterized serum samples: the Murdoch Childrens Research Institute Pneumococcal Laboratory (Melbourne, Australia) and the Vaccine Immunology Laboratory, National Public Health Institute (Helsinki, Finland). While good agreement was found for the inter-laboratory comparison for most serotypes, differences in ELISA methodology influenced specific IgG measurement. Therefore, use of the World Health Organization (WHO)-based ELISA methods for measurement of serotype-specific IgG is reliable, accurate and provides consistent results between international laboratories.

  17. Data-driven constraints on ice sheet model boundary conditions and comparison to borehole observations in Western Greenland

    Science.gov (United States)

    Meierbachtol, T. W.; Harper, J. T.; Johnson, J. V.; Humphrey, N. F.; Brinkerhoff, D. J.

    2013-12-01

    The utility of ice sheet models as prognostic tools relies on an accurate assessment of initial conditions. Ice sheet models reaching an initial state using assimilation techniques are inherently sensitive to the description of processes governing behavior at the ice-air and ice-bed boundaries. Propagation of uncertainty in these boundary condition effects exerts a strong control on the ice sheet thermal profile, which in turn impacts the basal thermal regime and partitioning of surface velocity into deformational and sliding components. With this in mind, correct implementation of boundary conditions when simulating ice flow is critical. Here, using the higher order numerical ice sheet model VarGlaS, we investigate the sensitivity of model output to field-based adjustments in surface and basal boundary conditions, using full thickness thermal profiles in the ice sheet ablation zone as a metric for comparison. Our measured temperature profiles provide a unique constraint by permitting evaluation of the integrated effect of necessary model assumptions and boundary conditions over long spatial scales greater than 100 km. We implement the study over a three-dimensional catchment of the Greenland ice sheet extending from the land terminating outlet glacier Isunnguata Sermia, east to the ice sheet divide. An initial reference case is generated from the surface and basal boundary fields of the SeaRise dataset. We then drive surface boundary changes using near-surface temperature measurements spanning 2 years in the ablation zone, and by scaling measurements of firn warming in western Greenland in the accumulation zone. Basal heat flux corrections follow direct measurement in a bedrock borehole adjacent to the study domain. Results show the downstream impact of substantial warming in the accumulation zone is limited by the ice sheet flow field, resulting in small changes to model temperatures in the vicinity of measured profiles.

  18. Comparison of the performance of IFA, CFA, and ELISA assays for the serodiagnosis of acute Q fever by quality assessment.

    Science.gov (United States)

    Herremans, Tineke; Hogema, Boris M; Nabuurs, Marrigje; Peeters, Marcel; Wegdam-Blans, Marjolijn; Schneeberger, Peter; Nijhuis, Carla; Notermans, Daan W; Galama, Joep; Horrevorts, Anton; van Loo, Inge H M; Vlaminckx, Bart; Zaaijer, Hans L; Koopmans, Marion P; Berkhout, Hanneke; Socolovschi, Cristina; Raoult, Didier; Stenos, John; Nicholson, William; Bijlmer, Henk

    2013-01-01

    The indirect immunofluorescence assay (IFA) is considered the reference method for diagnosing Q fever, but serology is also performed by complement fixation assay (CFA) or enzyme-linked immunosorbent assay (ELISA). However, comparability between these assays is not clear, and therefore a quality assessment was performed. A total of 25 serum samples from negative controls, Q fever patients, and a serial diluted high-positive sample were analyzed in 10 Dutch laboratories. Six laboratories performed CFA, 5 performed IFA, and 5 performed ELISAs. Three international reference laboratories from Australia, France, and the USA also participated in this study. Qualitative values between laboratories using the same methods were within close range, and all 3 methods correctly identified acute Q fever patients. The IFA, ELISA, and CFA are all suitable serodiagnostic assays to diagnose acute Q fever, but the IFA remains an important tool in the follow-up of patients and in identifying patients at risk for developing chronic Q fever. Copyright © 2013 Elsevier Inc. All rights reserved.

  19. Comparison of competitive ligand-binding assay and bioassay formats for the measurement of neutralizing antibodies to protein therapeutics.

    Science.gov (United States)

    Finco, Deborah; Baltrukonis, Daniel; Clements-Egan, Adrienne; Delaria, Kathy; Gunn, George R; Lowe, John; Maia, Mauricio; Wong, Teresa

    2011-01-25

    Administration of biological therapeutic proteins can lead to unwanted immunogenicity in recipients of these products. The assessment and characterization of such immune reactions can be helpful to better understand their clinical relevance and how they relate to patient safety and therefore, have become an integral part of a product development program for biological therapeutics. Testing for anti-drug antibodies (ADA) to biological/biotechnology-derived therapeutic proteins generally follows a tiered approach. Samples are initially screened for binding antibodies; presumptive positives are then confirmed in a confirmatory assay; subsequently, confirmed-positive samples may be further characterized by titration and with a neutralizing antibody (NAb) assay. Regulatory guidances on immunogenicity state that assessing the neutralizing capacity of antibodies should preferably be done using functional bioassays, while recognizing that competitive ligand-binding (CLB) assays may be substituted when neutralizing bioassays are inadequate or not feasible. This manuscript describes case studies from four companies in which CLB assays and functional bioassays were compared for their ability to detect neutralizing ADA against a variety of biotechnology-derived therapeutic proteins. Our findings indicate that CLB assays are comparable to bioassays for the detection of NAbs, in some cases offering better detection sensitivity, lower variability, and less matrix interference.

  20. Comparison of two multiplex PCR assays for the detection of Listeria spp. and Listeria monocytogenes in biological samples

    Directory of Open Access Journals (Sweden)

    Budniak Sylwia

    2016-12-01

    Full Text Available Introduction: The aim of the study was to optimise and compare two multiplex PCR assays for the detection of Listeria spp. and Listeria monocytogenes in biological samples including the liver, brain, and blood. Material and Methods: Three strains of L. monocytogenes and single strains of each of the species: L. ivanovii, L. innocua, L. grayi, L. welshimeri, and L. seeligeri were used. Additionally, five other species of bacterium were used to evaluate the specificity of the tests. Results: Specific amplification products were obtained for both multiplex PCR assays, which confirmed the tested strains as Listeria spp. and L. monocytogenes, respectively. Isolates of other species did not yield PCR products. Conclusion: Both multiplex PCR assays proved to be significantly sensitive and highly-specific methods for the detection of Listeria strains.

  1. Comparison of refractometry and biuret assay for measurement of total protein concentration in canine abdominal and pleural fluid specimens.

    Science.gov (United States)

    Rose, Alexandra; Funk, Deborah; Neiger, Reto

    2016-04-01

    To compare total protein (TP) concentrations in canine pleural and abdominal fluid specimens as measured by refractometry and biuret assay. Diagnostic test evaluation. Data regarding 92 pleural and 148 abdominal fluid specimens from dogs with various diseases. TP concentrations in fluid specimens as measured by refractometry and biuret assay were recorded. Strength of association between sets of measurements was assessed by Spearman rank correlations and Bland-Altman plots. Optimal concentration cutoff for diagnostic discrimination between exudate and nonexudate was identified by construction of receiver operating characteristic curves. Median TP concentration in pleural fluid specimens was 2.7 g/dL (range, 0.3 to 4.8 g/dL) for refractometry and 2.9 g/dL (range, 0.7 to 5.8 g/dL) for biuret assay. Median TP concentration in abdominal fluid specimens was 3.5 g/dL (range, 0.1 to 6.0 g/dL) for refractometry and 3.5 g/dL (range, 0.6 to 5.7 g/dL) for biuret assay. Correlation was significant between refractometric and biuret results for pleural (ρ = 0.921) and abdominal (ρ = 0.908) fluid. Bland-Altman plots revealed bias of -0.18 g/dL for pleural fluid and -0.03 g/dL for abdominal fluid for refractometry versus biuret assay. With a TP concentration of ≥ 3 g/dL used to distinguish exudate from nonexudate, sensitivity of refractometry was 77% for pleural fluid and 80% for abdominal fluid. Specificity was 100% and 94%, respectively. Refractometry yielded acceptable results for measurement of TP concentration in canine pleural and abdominal fluid specimens, providing a more rapid and convenient method than biuret assay.

  2. Comparison of Limulus assay, standard plate count, and total coliform count for microbiological assessment of renovated wastewater.

    Science.gov (United States)

    Jorgensen, J H; Lee, J C; Alexander, G A; Wolf, H W

    1979-05-01

    The Limulus endotoxin assay was compared to the standard plate count and total coliform count for assessment of the bacteriological quality of reclaimed wastewater. A total of 48 water samples from an advanced waste treatment plant in Dallas, Tex. were examined by the three techniques. Limulus assays were technically simpler to perform and provided results much sooner than conventional culture methods. However, the endotoxin values did not correlate extremely well with determinations of viable bacterial numbers. This lack of correlation may have been due to alterations in the normal ratio of viable gram-negative cells to endotoxin caused by water reclamation procedures.

  3. Comparison of Real-Time Multiplex Human Papillomavirus (HPV) PCR Assays with INNO-LiPA HPV Genotyping Extra Assay▿

    OpenAIRE

    Else, Elizabeth A.; Swoyer, Ryan; Zhang, Yuhua; Taddeo, Frank J.; Bryan, Janine T.; Lawson, John; Van Hyfte, Inez; Roberts, Christine C.

    2011-01-01

    Real-time type-specific multiplex human papillomavirus (HPV) PCR assays were developed to detect HPV DNA in samples collected for the efficacy determination of the quadrivalent HPV (type 6, 11, 16, and 18) L1 virus-like particle (VLP) vaccine (Gardasil). Additional multiplex (L1, E6, and E7 open reading frame [ORF]) or duplex (E6 and E7 ORF) HPV PCR assays were developed to detect high-risk HPV types, including HPV type 31 (HPV31), HPV33, HPV35, HPV39, HPV45, HPV51, HPV52, HPV56, HPV58, and H...

  4. A novel method for the determination of chemical purity and assay of menaquinone-7. Comparison with the methods from the official USP monograph.

    Science.gov (United States)

    Jedynak, Łukasz; Jedynak, Maria; Kossykowska, Magdalena; Zagrodzka, Joanna

    2017-02-20

    An HPLC method with UV detection and separation with the use of a C30 reversed phase analytical column for the determination of chemical purity and assay of menaquinone-7 (MK7) in one chromatographic run was developed. The method is superior to the methods published in the USP Monograph in terms of selectivity, sensitivity and accuracy, as well as time, solvent and sample consumption. The developed methodology was applied to MK7 samples of active pharmaceutical ingredient (API) purity, MK7 samples of lower quality and crude MK7 samples before purification. The comparison of the results revealed that the use of USP methodology could lead to serious overestimation (up to a few percent) of both purity and MK7 assay in menaquinone-7 samples. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Effects of currently used pesticides in the AhR-CALUX assay: comparison between the human TV101L and the rat H4IIE cell line

    DEFF Research Database (Denmark)

    Long, Manhai; Laier, Peter; Vinggaard, Anne Marie

    2003-01-01

    to be a rapid and sensitive assay for assessing the potency of AhR-activating compounds. We have used the AhR-CALUX assay to investigate the AhR-mediated activity of the persistent organochlorine insecticide dieldrin and twenty-two pesticides currently used in Denmark by employing the rat H4IIE and the human TV......101L hepatoma cell lines. In comparison the results indicated that the rat H4IIE cell line is more sensitive than the human TV101L for detection of TCDD inducing AhR-CALUX activity. The pesticides iprodione, chlorpyrifos and prochloraz showed dose-dependent AhR agonistic effects in both cell lines...... at concentrations above 10, 1 and 1 microM, respectively. However, some pesticides (methiocarb, chlorothalonil, tribenuron-methyl, paclobutrazol and tolchlofos-methyl) elicited differential responses in the two cell lines....

  6. Diagnostic accuracy and comparison of two assays for Borrelia-specific IgG and IgM antibodies

    DEFF Research Database (Denmark)

    Dessau, Ram

    2013-01-01

    Two assays (Liaison, Diasorin; IDEIA, Oxoid) for detection of Borrelia-specific antibodies were compared. A case-control design using patients with neuroborreliosis (n = 48), laboratory defined by a positive Borrelia-specific antibody index in the spinal fluid, was available and was intended...

  7. High performance liquid chromatography with two simultaneous on-line antioxidant assays: Evaluation and comparison of espresso coffees

    Energy Technology Data Exchange (ETDEWEB)

    Barnett, Neil W [ORNL; Gritti, Fabrice [University of Tennessee, Knoxville (UTK); Guiochon, Georges A [ORNL; Shalliker, R. Andrew [University of Western Sydney, Australia

    2010-01-01

    The antioxidant profiles of various espresso coffees were established using HPLC with UV-absorbance detection and two rapid, simultaneous, on-line chemical assays that enabled the relative reactivity of sample components to be screened. The assays were based on (i) the colour change associated with reduction of the 2,2'-diphenyl-1-picrylhydrazyl radical (DPPH{sm_bullet}); and (ii) the emission of light (chemiluminescence) upon reaction with acidic potassium permanganate. Results from the two approaches were similar and reflected the complex array of antioxidant species present in the samples. However, some differences in selectivity were observed. Chromatograms generated with the chemiluminescence assay contained more peaks, which was ascribed to the greater sensitivity of the reagent towards minor, readily oxidisable sample components. The three coffee samples produced closely related profiles, signifying their fundamentally similar chemical compositions and origin. Nevertheless, the overall intensity and complexity of the samples in both UV absorption and antioxidant assay chromatograms were aligned with the manufacturers description of flavour intensity and character.

  8. Comparison of interferon-gamma release assays and adenosine deaminase of pleural fluid for the diagnosis of pleural tuberculosis

    Institute of Scientific and Technical Information of China (English)

    刘菲

    2014-01-01

    Objective To compare the diagnostic performance of interferon gamma releasing assays(T-SPOT.TB)and adenosine deaminase(ADA)in pleural tuberculosis,and therefore to evaluate the value of T-SPOT.TB in a high tuberculosis burden country.Methods From June 2011to November 2012,111 patients with pleural fluid in Beijing Chest Hospital,Capital Medical University were

  9. Comparison of analytical and clinical performance of CLART HPV2 genotyping assay to Linear Array and Hybrid Capture 2

    DEFF Research Database (Denmark)

    Ejegod, Ditte Møller; Rebolj, Matejka; Bonde, Jesper

    2015-01-01

    prone to inter-observer variability. The reading of test results on the CLART HPV2 genotyping assay is, on the other hand, automated. The aim of our study was to directly compare the detection of HPV genotypes and high-grade cervical intraepithelial neoplasia (CIN) by CLART, Linear Array (LA...

  10. Determination of free insulin-like growth factor-I in human serum: comparison of ultrafiltration and direct immunoradiometric assay

    DEFF Research Database (Denmark)

    Frystyk, J; Ivarsen, P; Støving, R K;

    2001-01-01

    Two fundamentally different methods are currently used for the determination of free insulin-like growth factor-I (IGF-I): ultrafiltration by centrifugation (UF) and direct immunoradiometric assay (IRMA). The aim was to evaluate a commercial IRMA (DSL, Webster, TX, USA) and to compare it with UF....

  11. CONFIRMATIONAL IDENTIFICATION OF ESCHERICHIA COLI, A COMPARISON OF GENOTYPIC AND PHENOTYPIC ASSAYS FOR GLUTAMATE DECARBOXYLASE AND B-D-GLUCURONIDASE

    Science.gov (United States)

    Genotypic and phenotypic assays for glutamate decarboxylase (GAD) and B-D-glucuronidase (GUD) were compared for their abilities to detect various strains of Escherichia coli and to discriminate among other bacterial species. Test strains included nonpathogenic E.coli, three major...

  12. Direct comparison of Xpert MTB/RIF assay with liquid and solid mycobacterial culture for quantification of early bactericidal activity

    NARCIS (Netherlands)

    Kayigire, X.A.; Friedrich, S.O.; Venter, A.; Dawson, R.; Gillespie, S.H.; Boeree, M.J.; Heinrich, N.; Hoelscher, M.; Diacon, A.H.; Aarnoutse, R.

    2013-01-01

    The early bactericidal activity of antituberculosis agents is usually determined by measuring the reduction of the sputum mycobacterial load over time on solid agar medium or in liquid culture. This study investigated the value of a quantitative PCR assay for early bactericidal activity determinatio

  13. Transgenic Animal Mutation Assays

    Institute of Scientific and Technical Information of China (English)

    Tao Chen; Ph.D.D.A.B.T.

    2005-01-01

    @@ The novel transgenic mouse and rat mutation assays have provided a tool for analyzing in vivo mutation in any tissue, thus permitting the direct comparison of cancer incidence with mutant frequency.

  14. Comparison of mRNA splicing assay protocols across multiple laboratories: recommendations for best practice in standardized clinical testing.

    Science.gov (United States)

    Whiley, Phillip J; de la Hoya, Miguel; Thomassen, Mads; Becker, Alexandra; Brandão, Rita; Pedersen, Inge Sokilde; Montagna, Marco; Menéndez, Mireia; Quiles, Francisco; Gutiérrez-Enríquez, Sara; De Leeneer, Kim; Tenés, Anna; Montalban, Gemma; Tserpelis, Demis; Yoshimatsu, Toshio; Tirapo, Carole; Raponi, Michela; Caldes, Trinidad; Blanco, Ana; Santamariña, Marta; Guidugli, Lucia; de Garibay, Gorka Ruiz; Wong, Ming; Tancredi, Mariella; Fachal, Laura; Ding, Yuan Chun; Kruse, Torben; Lattimore, Vanessa; Kwong, Ava; Chan, Tsun Leung; Colombo, Mara; De Vecchi, Giovanni; Caligo, Maria; Baralle, Diana; Lázaro, Conxi; Couch, Fergus; Radice, Paolo; Southey, Melissa C; Neuhausen, Susan; Houdayer, Claude; Fackenthal, Jim; Hansen, Thomas Van Overeem; Vega, Ana; Diez, Orland; Blok, Rien; Claes, Kathleen; Wappenschmidt, Barbara; Walker, Logan; Spurdle, Amanda B; Brown, Melissa A

    2014-02-01

    Accurate evaluation of unclassified sequence variants in cancer predisposition genes is essential for clinical management and depends on a multifactorial analysis of clinical, genetic, pathologic, and bioinformatic variables and assays of transcript length and abundance. The integrity of assay data in turn relies on appropriate assay design, interpretation, and reporting. We conducted a multicenter investigation to compare mRNA splicing assay protocols used by members of the ENIGMA (Evidence-Based Network for the Interpretation of Germline Mutant Alleles) consortium. We compared similarities and differences in results derived from analysis of a panel of breast cancer 1, early onset (BRCA1) and breast cancer 2, early onset (BRCA2) gene variants known to alter splicing (BRCA1: c.135-1G>T, c.591C>T, c.594-2A>C, c.671-2A>G, and c.5467+5G>C and BRCA2: c.426-12_8delGTTTT, c.7988A>T, c.8632+1G>A, and c.9501+3A>T). Differences in protocols were then assessed to determine which elements were critical in reliable assay design. PCR primer design strategies, PCR conditions, and product detection methods, combined with a prior knowledge of expected alternative transcripts, were the key factors for accurate splicing assay results. For example, because of the position of primers and PCR extension times, several isoforms associated with BRCA1, c.594-2A>C and c.671-2A>G, were not detected by many sites. Variation was most evident for the detection of low-abundance transcripts (e.g., BRCA2 c.8632+1G>A Δ19,20 and BRCA1 c.135-1G>T Δ5q and Δ3). Detection of low-abundance transcripts was sometimes addressed by using more analytically sensitive detection methods (e.g., BRCA2 c.426-12_8delGTTTT ins18bp). We provide recommendations for best practice and raise key issues to consider when designing mRNA assays for evaluation of unclassified sequence variants.

  15. A comparison of intradermal testing and detection of allergen-specific immunoglobulin E in serum by enzyme-linked immunosorbent assay in horses affected with skin hypersensitivity.

    Science.gov (United States)

    Morgan, Erin E; Miller, William H; Wagner, Bettina

    2007-12-15

    Skin hypersensitivities (allergies) in horses are often diagnosed using clinical signs only. Intradermal testing or serological assays are diagnostic options to confirm the allergic nature of the disease and to identify the allergen(s). Our objective was to develop an allergen-specific enzyme-linked immunosorbent assay (ELISA) using a monoclonal antibody specific for horse IgE and to examine its potential for allergen detection in serum in comparison to intradermal testing. Intradermal testing with 61 allergen extracts was performed on 10 horses affected with skin hypersensitivity. Their sera were analyzed by ELISA for IgE antibodies to the same allergens. The kappa test of concordance was used for comparison of the results of both tests. Out of 61 allergen extracts, only two (Timothy and Quack) had kappa values greater than 0.60, suggesting a substantial agreement between skin testing and IgE ELISA. The statistical comparison of the remaining 59 allergens showed little or no concordance between the tests beyond chance. To identify parameters that may influence the sensitivity of the ELISA, the assay was modified to detect allergen-specific IgGb and IgG(T) in serum, and the protein content in all allergen extracts was determined by SDS-PAGE. The commercial allergen extracts revealed a high variation in detectable protein. High concentrations of allergen-specific IgG in horse serum were found to compete with IgE for binding to the plates. In conclusion, an ELISA using whole serum and crude allergen preparations provides limited diagnostic information in horses. The reliable diagnosis of allergens in equine skin hypersensitivity is essential to improve allergen-specific treatments, such as hyposensitization, or the development of allergy vaccines.

  16. A comparison of the analytical level of agreement of nine treponemal assays for syphilis and possible implications for screening algorithms.

    Science.gov (United States)

    Jost, Heather; Castro, Arnold; Cox, David; Fakile, Yetunde; Kikkert, Susan; Tun, Ye; Zaidi, Akbar; Park, Mahin

    2013-09-19

    The serological diagnosis of syphilis requires the detection of two distinct antibodies, the non-treponemal and trepomenal. Center for Disease Control and Prevention (CDC) recommends screening first with a non-treponemal test such as (Rapid Plasma Reagin/Venereal Disease Research Laboratory), and then confirming those results with one of the several treponemal tests (Fluorescent Treponemal Antibody-Absorption (FTA-ABS), Enzyme Immunoassay, chemiluminescence, treponema pallidum particle agglutination (TP-PA) or Point of Care). Owing to the high volume of samples processed by some laboratories using automated systems, the screening with treponemal assays and confirming with non-treponemal tests is becoming the established norm. The purpose of this study was to evaluate eight treponemal assays using TP-PA as the predicate assay. 290 stored serum samples were tested qualitatively according to the manufacturer's directions. Concordance with specimens tested as reactive or non-reactive using TP-PA was: FTA-ABS 94.5-100%, Trep-Sure 100-98.9%, BioELISA 100-98.9%, INNO-LIA 99.1-99.4%, BIOLINE 100-98.9%, CAPTIA IgG 100-97.2%, Trep-ID 100-100% and LIAISON 100-99.4%. In order to properly evaluate the performance of these assays, the analytical sensitivity was determined by endpoint titration of serial dilutions of the reactive serum samples in normal sera. The median endpoint titre varied from 1:4 for FTA-ABS to 1:512 for Trep-Sure. The performance of the treponemal serological assays was comparable while using medium and high-titre sera. However, the varying performance on specimen dilutions suggests that there may be differences in sensitivity with low-titre sera that are more prevalent in primary and late syphilis cases.

  17. Comparison of nucleic acid amplification assays with BD affirm VPIII for diagnosis of vaginitis in symptomatic women.

    Science.gov (United States)

    Cartwright, Charles P; Lembke, Bryndon D; Ramachandran, Kalpana; Body, Barbara A; Nye, Melinda B; Rivers, Charles A; Schwebke, Jane R

    2013-11-01

    A commercially available, nonamplified, nucleic acid probe-based test system (BD Affirm VPIII) was compared with nucleic acid amplification (NAA)-based assays for determining the etiology of vaginitis in a cohort of 323 symptomatic women. First, a semiquantitative, multiplexed PCR assay (BV-PCR) and the Affirm VPIII Gardnerellavaginalis test were compared with a unified bacterial-vaginosis (BV) reference standard incorporating both Nugent Gram stain scores and Amsel clinical criteria. In the evaluable population of 305 patients, BV-PCR was 96.9% (191/197) sensitive and 92.6% specific (100/108) for BV, while Affirm VPIII was 90.1% sensitive (179/197) and 67.6% specific (73/108). Second, a multiplexed PCR assay detecting Candida albicans and Candida glabrata (CAN-PCR) was compared with the Affirm VPIII Candida test using a reference standard for vulvovaginal candidiasis (VVC) of yeast culture plus exclusion of alternate vaginitis etiologies. In the population evaluated (n = 102), CAN-PCR was 97.7% sensitive (42/43) and 93.2% specific (55/59) and Affirm VP III was 58.1% sensitive (25/43) and 100% specific (59/59) for VVC. Finally, the results of a commercial NAA test (GenProbe Aptima Trichomonas vaginalis assay; ATV) for T. vaginalis were compared with the Affirm VPIII Trichomonas vaginalis test. In the absence of an independent reference standard for trichomonal vaginitis (TV), a positive result in either assay was deemed to represent true infection. In the evaluable cohort of 388 patients, the sensitivity of ATV was 98.1% (53/54) versus 46.3% (25/54) for Affirm VPIII. The diagnostic accuracy of the combined NAA-based test construct was approximately 20 to 25% higher than that of the Affirm VPIII when modeled in populations with various prevalences of infectious vaginitis.

  18. Effects of currently used pesticides in the AhR-CALUX assay: comparison between the human TV101L and the rat H4IIE cell line

    DEFF Research Database (Denmark)

    Long, M.; Laier, Peter; Vinggaard, Anne;

    2003-01-01

    The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that mediates many of the biologic and toxicological effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and related compounds. The in vitro chemically activated luciferase expression (CALUX) assay has been proven...... TV101L hepatoma cell lines. In comparison the results indicated that the rat H4IIE cell line is more sensitive than the human TV101L for detection of TCDD inducing AhR-CALUX activity. The pesticides iprodione, chlorpyrifos and prochloraz showed dose-dependent AhR agonistic effects in both cell lines...

  19. A review of abortion laws in Western-European countries. A cross-national comparison of legal developments between 1960 and 2010.

    Science.gov (United States)

    Levels, Mark; Sluiter, Roderick; Need, Ariana

    2014-10-01

    The extent to which women have had access to legal abortions has changed dramatically in Western-Europe between 1960 and 2010. In most countries, abortion laws developed from completely banning abortion to allowing its availability on request. Both the timing and the substance of the various legal developments differed dramatically between countries. Existing comparative studies on abortion laws in Western-European countries lack detail, usually focus either on first-trimester abortions or second trimester abortions, cover a limited time-span and are sometimes inconsistent with one another. Combining information from various primary and secondary sources, we show how and when the conditions for legally obtaining abortion during the entire gestation period in 20 major Western-European countries have changed between 1960 and 2010. We also construct a cross-nationally comparable classification of procedural barriers that limit abortion access. Our cross-national comparison shows that Western-Europe witnessed a general trend towards decreased restrictiveness of abortion laws. However, legal approaches to regulating abortion are highly different in detail. Abortion access remains limited, sometimes even in countries where abortion is legally available without restrictions relating to reasons. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  20. Identification of Giardia lamblia and the human infectious-species of Cryptosporidium in drinking water resources in Western Saudi Arabia by nested-PCR assays.

    Science.gov (United States)

    Hawash, Y; Ghonaim, M; Hussein, Y; Alhazmi, A; Alturkistani, A

    2015-06-01

    The presence of Cryptosporidium and/or Giardia in drinking water represents a major public health problem. This study was the first report concerned with the occurrence of these protozoa in drinking water in Saudi Arabia. The study was undertaken in Al-Taif, a high altitude region, Western Saudi Arabia. Eight underground wells water, six desalinated water and five domestic brands of bottled water samples, 10 liter each, were monthly collected between May 2013 and April 2014. All samples (n = 228), were processed using an automated wash/elution station (IDEXX Laboratories, Inc.). Genomic DNA was directly isolated and purified from samples concentrates with QIAamp® Stool Mini Kit (Qiagen). The target protozoan DNA sequences were amplified using two previously published nested-PCR protocols. Of all the analyzed water, 31 samples (≈14%) were found contaminated with the target protozoa. Giardia lamblia was detected in ≈10% (7/72) of desalinated water and in ≈9% (9/96) of wells water. On the other hand, Cryptosporidium was identified in ≈8% (8/72) of desalinated water and in ≈7% (7/96) of wells water. All bottled water samples (n = 60) were (oo)cysts-free. Protozoan (oo)cysts were more frequently identified in water samples collected in the spring than in other seasons. The methodology established in our study proved sensitive, cost-effective and is amenable for future automation or semi-automation. For better understanding of the current situation that represent an important health threat to the local inhabitants, further studies concerned with (oo)cyst viability, infectivity, concentration and genotype identification are recommended.

  1. Evaluation of the dot enzyme-linked immunosorbent assay in comparison with standard ELISA for the immunodiagnosis of human toxocariasis

    Directory of Open Access Journals (Sweden)

    Roldán William

    2006-01-01

    Full Text Available A dot enzyme-linked immunosorbent assay (dot-ELISA was standardized using excretory-secretory antigens of Toxocara canis for the rapid immunodiagnosis of human toxocariasis. Thirty patients with clinical signs of toxocariasis, 20 cases with other parasitic diseases, and 40 healthy subjects were tested. A total of 0.2 ng of antigen per dot, serum dilution of 1:160 and dilution conjugate of 1:1000 were found optimal. The sensitivity and specificity of the assay were 100 and 95%, respectively. Comparable sensitivity of dot-ELISA and the standard ELISA was obtained, but only 3 cross-reactions occurred in the dot-ELISA, compared with 6 in the standard ELISA. Dot-ELISA is simple to perform, rapid, and low cost. Large-scale screening studies should be done to evaluate its usefulness under field conditions.

  2. Comparison of solid-phase and eluate assays to gauge the ecotoxicological risk of organic wastes on soil organisms.

    Science.gov (United States)

    Domene, Xavier; Alcañiz, Josep M; Andrés, Pilar

    2008-02-01

    Development of methodologies to assess the safety of reusing polluted organic wastes in soil is a priority in Europe. In this study, and coupled with chemical analysis, seven organic wastes were subjected to different aquatic and soil bioassays. Tests were carried out with solid-phase waste and three different waste eluates (water, methanol, and dichloromethane). Solid-phase assays were indicated as the most suitable for waste testing not only in terms of relevance for real situations, but also because toxicity in eluates was generally not representative of the chronic effects in solid-phase. No general correlations were found between toxicity and waste pollutant burden, neither in solid-phase nor in eluate assays, showing the inability of chemical methods to predict the ecotoxicological risks of wastes. On the contrary, several physicochemical parameters reflecting the degree of low organic matter stability in wastes were the main contributors to the acute toxicity seen in collembolans and daphnids.

  3. Pulmonary toxicity of nanomaterials: a critical comparison of published in vitro assays and in vivo inhalation or instillation studies.

    Science.gov (United States)

    Landsiedel, Robert; Sauer, Ursula G; Ma-Hock, Lan; Schnekenburger, Jürgen; Wiemann, Martin

    2014-11-01

    To date, guidance on how to incorporate in vitro assays into integrated approaches for testing and assessment of nanomaterials is unavailable. In addressing this shortage, this review compares data from in vitro studies to results from in vivo inhalation or intratracheal instillation studies. Globular nanomaterials (ion-shedding silver and zinc oxide, poorly soluble titanium dioxide and cerium dioxide, and partly soluble amorphous silicon dioxide) and nanomaterials with higher aspect ratios (multiwalled carbon nanotubes) were assessed focusing on the Organisation for Economic Co-Operation and Development (OECD) reference nanomaterials for these substances. If in vitro assays are performed with dosages that reflect effective in vivo dosages, the mechanisms of nanomaterial toxicity can be assessed. In early tiers of integrated approaches for testing and assessment, knowledge on mechanisms of toxicity serves to group nanomaterials thereby reducing the need for animal testing.

  4. Genomewide Linkage Analysis of Bipolar Disorder by Use of a High-Density Single-Nucleotide–Polymorphism (SNP) Genotyping Assay: A Comparison with Microsatellite Marker Assays and Finding of Significant Linkage to Chromosome 6q22

    Science.gov (United States)

    Middleton, F. A.; Pato, M. T.; Gentile, K. L.; Morley, C. P.; Zhao, X.; Eisener, A. F.; Brown, A.; Petryshen, T. L.; Kirby, A. N.; Medeiros, H.; Carvalho, C.; Macedo, A.; Dourado, A.; Coelho, I.; Valente, J.; Soares, M. J.; Ferreira, C. P.; Lei, M.; Azevedo, M. H.; Kennedy, J. L.; Daly, M. J.; Sklar, P.; Pato, C. N.

    2004-01-01

    We performed a linkage analysis on 25 extended multiplex Portuguese families segregating for bipolar disorder, by use of a high-density single-nucleotide–polymorphism (SNP) genotyping assay, the GeneChip Human Mapping 10K Array (HMA10K). Of these families, 12 were used for a direct comparison of the HMA10K with the traditional 10-cM microsatellite marker set and the more dense 4-cM marker set. This comparative analysis indicated the presence of significant linkage peaks in the SNP assay in chromosomal regions characterized by poor coverage and low information content on the microsatellite assays. The HMA10K provided consistently high information and enhanced coverage throughout these regions. Across the entire genome, the HMA10K had an average information content of 0.842 with 0.21-Mb intermarker spacing. In the 12-family set, the HMA10K-based analysis detected two chromosomal regions with genomewide significant linkage on chromosomes 6q22 and 11p11; both regions had failed to meet this strict threshold with the microsatellite assays. The full 25-family collection further strengthened the findings on chromosome 6q22, achieving genomewide significance with a maximum nonparametric linkage (NPL) score of 4.20 and a maximum LOD score of 3.56 at position 125.8 Mb. In addition to this highly significant finding, several other regions of suggestive linkage have also been identified in the 25-family data set, including two regions on chromosome 2 (57 Mb, NPL = 2.98; 145 Mb, NPL = 3.09), as well as regions on chromosomes 4 (91 Mb, NPL = 2.97), 16 (20 Mb, NPL = 2.89), and 20 (60 Mb, NPL = 2.99). We conclude that at least some of the linkage peaks we have identified may have been largely undetected in previous whole-genome scans for bipolar disorder because of insufficient coverage or information content, particularly on chromosomes 6q22 and 11p11. PMID:15060841

  5. Comparison of a rapid ATP bioluminescence assay and standard plate count methods for assessing microbial contamination of consumers' refrigerators.

    Science.gov (United States)

    Chen, Fur-Chi; Godwin, Sandria L

    2006-10-01

    The feasibility of using an ATP bioluminescence assay for assessing microbial contamination of home refrigerators was evaluated and compared with the standard culture methods. Samples of refrigerator surfaces were collected from 123 households by swabbing an area of 100 cm2 on three locations in the refrigerator with premoisturized sterile swabs. Microbial contaminations were determined by aerobic plate count (APC; incubated at 35 degrees C for 48 h) and psychrotrophic plate count (PPC; incubated at 7 degrees C for 10 days) on plate count agar. The results were compared to the readings from the microbial ATP (mATP) bioluminescence assay. The correlation coefficient (r) between mATP and PPC (r = 0.851) was slightly higher than that between mATP and APC (r = 0.823). Our results indicated a potential discrepancy in the population of mesophilic and psychrotrophic bacteria in the refrigerator samples. Nevertheless, mATP appeared to be a reliable indication of the average of APC and PPC (r = 0.895). The mATP bioluminescence assay would provide a rapid and convenient test for researchers in field studies to assess microbial contamination in refrigerators.

  6. Comparison of ante-mortem assays to assess progression/regression of paratuberculosis in individual dairy animals.

    Science.gov (United States)

    Click, Robert E; Van Kampen, Craig L

    2010-01-01

    Johne's disease, caused by Mycobacterium avium, subspecies paratuberculosis (MAP) is becoming increasingly widespread on dairy farms worldwide, due in part, to the absence of vaccine/drug or curative modalities.  This spread is of concern since MAP is at the center of a controversy as to its role in Crohn's disease.  None of the methods presently available to define paratuberculosis in cattle have been examined for their ability to assess progression/regression of any treatment or intervention of this disease   The research presented herein, therefore was designed to assess the reliability and accuracy of available ante-mortem assays to predict disease change of individual animals undergoing a probiotic, potentially therapeutic, treatment.  Paratuberculosis positive (n = 75) and negative (n = 10) animals were longitudinally monitored over their natural lifetimes with specific serum antibody and fecal shedding assays, and for development of end-stage clinical disease.  Longitudinal, increasing/decreasing serum ELISA values were associated with, and predictive of, progression/regression of disease.  Changes in fecal shedding and serum AGID were of value at only specific stages.  Documentation that ELISA-positive animals were positive for paratuberculosis was done by a compilation of ELISA-independent assays--succumbing with end-stage clinical disease, autopsy, AGID, and MAP fecal shedding.

  7. Diagnosis of preeclampsia with soluble Fms-like tyrosine kinase 1/placental growth factor ratio: an inter-assay comparison.

    Science.gov (United States)

    Andersen, Louise Bjørkholt; Frederiksen-Møller, Britta; Work Havelund, Kathrine; Dechend, Ralf; Jørgensen, Jan Stener; Jensen, Boye L; Nielsen, Jan; Lykkedegn, Sine; Barington, Torben; Christesen, Henrik Thybo

    2015-02-01

    The angiogenic factor ratio soluble Fms-kinase 1 (sFlt-1)/placental growth factor (PlGF) is a novel diagnostic tool for preeclampsia. We compared the efficacy of the KRYPTOR (BRAHMS) automated assays for sFlt-1 and PlGF with the Elecsys (Roche) assays in a routine clinical setting. Preeclamptic women (n = 39) were included shortly after the time of diagnosis. Normotensive control pregnancies were matched by gestational age (n = 76). The KRYPTOR assays performed comparably or superior to Elecsys (sFlt-1/PlGF area under the curve 0.746 versus 0.735; P = .09; for non-obese 0.820 versus 0.805, P = .047). For early-onset preeclampsia, KRYPTOR area under the curve increased to 0.929 with a 100% specificity for preeclampsia at cut-off 85 and an 88.9% sensitivity for preeclampsia at cut-off 33. For women with preeclampsia and preterm delivery or Hemolysis, Elevated Liver enzymes, Low Platelet count (HELLP) syndrome, the KRYPTOR sFlt-1/PlGF ratio was manifold increased (P < .01). The sFlt-1/PlGF ratio proved especially useful in early-onset preeclampsia, preeclampsia with preterm delivery or HELLP, and among non-obese women.

  8. Comparison of targeted peptide quantification assays for reductive dehalogenases by selective reaction monitoring (SRM) and precursor reaction monitoring (PRM).

    Science.gov (United States)

    Schiffmann, Christian; Hansen, Rasmus; Baumann, Sven; Kublik, Anja; Nielsen, Per Halkjær; Adrian, Lorenz; von Bergen, Martin; Jehmlich, Nico; Seifert, Jana

    2014-01-01

    Targeted absolute protein quantification yields valuable information about physiological adaptation of organisms and is thereby of high interest. Especially for this purpose, two proteomic mass spectrometry-based techniques namely selective reaction monitoring (SRM) and precursor reaction monitoring (PRM) are commonly applied. The objective of this study was to establish an optimal quantification assay for proteins with the focus on those involved in housekeeping functions and putative reductive dehalogenase proteins from the strictly anaerobic bacterium Dehalococcoides mccartyi strain CBDB1. This microbe is small and slow-growing; hence, it provides little biomass for comprehensive proteomic analysis. We therefore compared SRM and PRM techniques. Eleven peptides were successfully quantified by both methods. In addition, six peptides were solely quantified by SRM and four by PRM, respectively. Peptides were spiked into a background of Escherichia coli lysate and the majority of peptides were quantifiable down to 500 amol absolute on column by both methods. Peptide quantification in CBDB1 lysate resulted in the detection of 15 peptides using SRM and 14 peptides with the PRM assay. Resulting quantification of five dehalogenases revealed copy numbers of <10 to 115 protein molecules per cell indicating clear differences in abundance of RdhA proteins during growth on hexachlorobenzene. Our results indicated that both methods show comparable sensitivity and that the combination of the mass spectrometry assays resulted in higher peptide coverage and thus more reliable protein quantification.

  9. A Comparison of Nested PCR Assay with Conventional Techniques for Diagnosis of Intestinal Cryptosporidiosis in AIDS Cases from Northern India

    Directory of Open Access Journals (Sweden)

    Beena Uppal

    2014-01-01

    Full Text Available Cryptosporidiosis is a very important opportunistic infection and is responsible for significant morbidity and mortality in HIV/AIDS patients. Although current laboratory methods are generally considered adequate to detect high concentrations of oocysts, they fail to detect cases of cryptosporidiosis in many immunocompromised patients. The present study was done to determine the diagnostic efficacy of modified Ziehl-Neelsen (ZN, antigen detection ELISA, and a nested PCR assay for detection of Cryptosporidium in 58 adult AIDS cases with diarrhea from the ART clinic of Lok Nayak Hospital, New Delhi. Cryptosporidium was detected in 17 (29.4%, 39 (67.3%, and 45 (77.5% cases by modified ZN staining, antigen ELISA, and nested PCR assay, respectively. Taking nested PCR as the gold standard, specificity of both modified ZN staining and Cryptosporidium antigen detection ELISA was 100% while the sensitivity of the tests was 37.8% and 86.6%, respectively. PCR was more sensitive than the other two diagnostic modalities but required a more hands-on time per sample and was more expensive than microscopy. PCR, however, was very adaptable to batch analysis, reducing the costs considerably. This assay can therefore have considerable advantages in the treatment of immunosuppressed individuals like AIDS patients, allowing their early diagnosis and decreasing the morbidity and the mortality.

  10. Comparison of two molecular assays for detection of cytomegalovirus DNA in whole blood and plasma samples from transplant recipients.

    Science.gov (United States)

    Costa, Cristina; Sidoti, Francesca; Mantovani, Samantha; Gregori, Gabriella; Proietti, Alex; Ghisetti, Valeria; Cavallo, Rossana

    2016-07-01

    In immunosuppressed patients, pre-emptive therapy and a strict follow-up of CMV infection are the standard of care for the prevention of CMV disease. Several real-time PCR assays for CMV DNA quantification on whole blood (WB) and plasma (PL) are commercially available. This study compared and correlated CMV viral loads obtained by the Cobas AmpliPrep/Cobas TaqMan (CAP/CTM) platform on plasma specimens with those obtained on corresponding whole blood specimens by the real-time PCR assay (ELITe MGB-CMV) in 185 sequential samples from 41 immunosuppressed patients. Correlation between the two assays was good. Kinetics of CMV DNA within the same patient was similar, but PL viral load was constantly 1 log lower than WB. In patients under antiviral therapy, low level of CMV DNA persisted in WB, while it was absent in PL. The good correlation between CMV DNA detected on both PL and WB supports the reliability of the two matrices for viral monitoring and the therapeutic management of CMV infection. Nevertheless, due to significant quantification differences between PL and WB CMV DNA, the same biological specimen should be used for a sequential and reliable follow-up of patients at high risk of CMV infection.

  11. Comparison of nested and ELISA based polymerase chain reaction assays for detecting Chlamydia trachomatis in pregnant women with preterm complications.

    Science.gov (United States)

    Sulaiman, S; Chong, P P; Mokhtarudin, R; Lye, M S; Wan Hassan, W H

    2014-03-01

    Identification of pregnant women infected with Chlamydia trachomatis is essential to allow early antibiotic treatment in order to prevent adverse pregnancy outcomes. In this study, two nucleic acid amplification tests (NAAT) namely nested PCR (BioSewoom, Korea) and Amplicor CT/NG (Roche Diagnostic, USA) were evaluated in terms of sensitivity and specificity for the detection of C. trachomatis DNA in pregnant women with preterm complications. A cross-sectional study was carried out in two public hospitals in Southern Selangor, Malaysia. Endocervical swabs obtained were subjected to DNA amplification using nested PCR (BioSewoom, Korea) and Amplicor CT/NG (Roche Diagnostic, USA). A total of 83 endocervical swabs obtained from pregnant women of less than 37 weeks gestation and presented with preterm complications were subjected to chlamydial DNA detection using both assays. The study shows that Amplicor CT/NG assay is more effective in the detection of C. trachomatis DNA from endocervical swabs compared to Biosewoom nested PCR kit. Agreement between the two assays were poor (kappa=0.094) with nested PCR showing a low sensitivity of 10.81% and a 97.83% specificity when compared to Amplicor CT/NG. The results obtained indicated that BioSewoom nested PCR was less sensitive than Amplicor CT/ NG for detecting C. trachomatis in endocervical specimens and that another more reliable test is required for confirmatory result.

  12. Anti-glomerular basement membrane antibodies in the diagnosis of Goodpasture syndrome: a comparison of different assays.

    Science.gov (United States)

    Sinico, Renato Alberto; Radice, Antonella; Corace, Caterina; Sabadini, Ettore; Bollini, Bruna

    2006-02-01

    The role of anti-glomerular basement membrane (GBM) antibodies in the pathogenesis of Goodpasture syndrome (GPS) is firmly established. Untreated, the disease may follow a fulminating course. Early identification of patients has important implications in terms of management and prognosis. Therefore, a diagnostic test for the determination of circulating anti-GBM antibodies, of very high sensitivity and specificity, is necessary. A number of assays, using different antigenic substrates, are available, but studies comparing the 'performances' of the different tests are scarce. The aim of our work was to evaluate the sensitivity and specificity of four immunoassay-based anti-GBM antibodies kits. Thirty-four serum samples from 19 GPS patients, 41 pathological and 28 normal controls were studied retrospectively (the follow-up samples were not included in the analysis of performance data). Cut-off limits were derived from receiver operating characteristics curve analysis. All the assays showed a comparable good sensitivity (between 94.7 and 100.0%), whereas specificity varied considerably (from 90.9 to 100.0%). The better performance in terms of sensitivity/specificity was achieved by a fluorescence immunoassay which utilizes a recombinant antigen. All the assays have a good performance, with high sensitivity; however, the specificity may vary considerably.

  13. Spatial-temporal analysis of HIV-1 PR and RT resistance-associated mutations of nucleotide sequences from Western Europe, using vircoTYPE™ HIV-1 assay

    Directory of Open Access Journals (Sweden)

    M Govaert

    2012-11-01

    , with a SR of 59% & RR of 41% for both drugs. For NRTIs, the highest SR was found for stavudine (77%, 20889/27262 followed by tenofovir (67%, 18249/27262 with 23% (6499/27262 resistant sequences observed for stavudine & 33% (9114/27262 for tenofovir. The current analysis provides some preliminary insight into HIV mutation pattern prevalence and resistance within Western Europe, suggesting good therapeutic opportunities for regimens containing new generation PIs & NNRTIs.

  14. The Effect of Spatial Scale on Paleovegetation Data-Model Comparisons for 6 ka and 21 ka in the Western United States

    Science.gov (United States)

    Shafer, S. L.; Bartlein, P. J.; Thompson, R. S.; Strickland, L. E.

    2010-12-01

    Climate-model simulations of past climates are often evaluated using both paleovegetation proxy data (e.g., pollen) and model simulations of past vegetation. These data-model comparison efforts provide important information for evaluating paleoclimate model simulations, and also for refining interpretations of paleovegetation proxy data and improving our understanding of past climate-vegetation interactions. A common limitation of these data-model comparisons is that the spatial resolution of paleoclimate simulations is frequently coarser than the spatial scales represented by the vegetation proxy data. This difference in spatial resolution is particularly important in topographically complex regions, such as the western United States. We used climate data simulated by coupled atmosphere-ocean general circulation models for 6 ka and 21 ka from the Paleoclimate Modelling Intercomparison Project phase 2 (PMIP2) database to evaluate the effects of spatial scale on the agreement between simulated and observed paleovegetation data. The climate data were downscaled to spatial resolutions of 30-minutes and finer and used with BIOME4, an equilibrium biogeography model, to simulate equilibrium vegetation for the western United States. The simulated vegetation was compared with fossil pollen and macrofossil data for the same region from the BIOME6000 data set. The results indicate the sensitivity of different vegetation types to changes in the spatial resolution of the data-model comparison and demonstrate the importance of data aggregation and spatial scale in evaluating data-model agreement.

  15. Why, when, and how to diversify? A comparison between Western theories and the cognition of Chinese enterprises

    Institute of Scientific and Technical Information of China (English)

    JIA Liangding; ZHANG Junjun; QIAN Haiyan; CUI Rongjun; CHEN Yongxia

    2007-01-01

    This paper uses as research samples 140 papers on enterprise diversification published in top-notch Western journals, and public statements from 30 influential contemporary Chinese CEOs on enterprise diversification. Both the qualitative open coding and the qualitative factor analysis are employed to analyze the two samples respectively, and then the corresponding analysis is utilized to explore the differences between Western theories and the cognition of Chinese enterprises on the motivation (why), timing (when) and industry choice (how) of enterprise diversification. Results show that, first, both consider the motivation of diversification mainly from the perspectives of resource-based view and asset portfolio theory. However, Western theories pay more attention to the factors related to the perspectives of the resource-based theory, transaction cost theory and agency theory, while Chinese enterprises put more emphasis on those factors associated with the asset portfolio theory, government policies and institutional theory. Second, on the cognition of the timing of diversification, Western theories insist that enterprises should diversify when they meet threats, while the practice of Chinese enterprises insists that diversification should take place when enterprises have enough strength. Third, Western theories focus more on the interrelationship between the original industry and the intended industry than on the attractiveness of the intended industry, while Chinese enterprises pay more attention to attractiveness than interrelationship.

  16. Baseline morning cortisol level as a predictor of pituitary-adrenal reserve: a comparison across three assays.

    Science.gov (United States)

    Sbardella, Emilia; Isidori, Andrea M; Woods, Conor P; Argese, Nicola; Tomlinson, Jeremy W; Shine, Brian; Jafar-Mohammadi, Bahram; Grossman, Ashley B

    2017-02-01

    The short ACTH stimulation test (250 μg) is the dynamic test most frequently used to assess adrenal function. It is possible that a single basal cortisol could be used to predict the dynamic response, but research has been hampered by the use of different assays and thresholds. To propose a morning baseline cortisol criterion of three of the most commonly used modern cortisol immunoassays - Advia Centaur (Siemens), Architect (Abbott) and the Roche Modular System (Roche) - that could predict adrenal sufficiency. Observational, retrospective cross-sectional study at two centres. Retrospective analysis of the results of 1019 Short Synacthen tests (SSTs) with the Advia Centaur, 449 SSTs with the Architect and 2050 SSTs with the Roche Modular System assay. Serum cortisol levels were measured prior to injection of 250 μg Synacthen and after 30 min. Overall, we were able to collate data from a total of 3518 SSTs in 3571 patients. Using receiver-operator curve analysis, baseline cortisol levels for predicting passing the SST with 100% specificity were 358 nmol/l for Siemens, 336 nmol/l for Abbott and 506 nmol/l for Roche. Utilizing these criteria, 589, 158 and 578 SSTs, respectively, for Siemens, Abbott and Roche immunoassays could have been avoided. We have defined assay-specific morning cortisol levels that are able to predict the integrity of the hypothalamo-pituitary-adrenal axis. We propose that this represents a valid tool for the initial assessment of adrenal function and has the potential to obviate the need for dynamic testing in a significant number of patients. © 2016 John Wiley & Sons Ltd.

  17. Comparison of real-time PCR and antigen assays for detection of hepatitis E virus in blood donors.

    Science.gov (United States)

    Vollmer, T; Knabbe, C; Dreier, J

    2014-06-01

    Hepatitis E virus (HEV) infection is recognized as an emerging and often undiagnosed disease in industrialized countries, with asymptomatic infections actually occurring in blood donors. Sensitive detection of HEV-RNA is crucial for diagnosis and monitoring of disease progression. We evaluated the analytical sensitivity and performance of three HEV RT-PCR assays (RealStar HEV reverse transcription-PCR [RT-PCR], hepatitis@ceeramTools, and ampliCube HEV RT-PCR) for screening of individuals for HEV infections (ID-nucleic acid amplification technology [ID-NAT]) and for blood donor pool screening (minipool-NAT [MP-NAT]). RNA was extracted using NucliSens easyMAG (ID-NAT) and a high-volume extraction protocol (4.8 ml, chemagic Viral 5K, MP-NAT). Three NAT assays were evaluated for ID-NAT but only two assays for MP-NAT due to inhibition of the ampliCube HEV RT-PCR kit using the corresponding RNA extract. Assays provided good analytical sensitivity, ranging from 37.8 to 180.1 IU/ml (ID-NAT) and from 4.7 to 91.2 IU/ml (MP-NAT). The applicability of HEV antigen (HEV-Ag) screening was compared to that of RT-PCR screening and detection of HEV-IgM antibodies using seroconversion panels of 10 HEV genotype 3-infected individuals. Four individuals revealed a positive HEV-Ag detection result, with corresponding viremias ranging from 1.92 E + 03 to 2.19 E + 05 IU/ml, while the progression of HEV-Ag followed that of HEV viremia. The other six individuals showed no presence of HEV-Ag although the corresponding viremias were also in the range of >1.0 E + 03. Anti-HEV-IgM antibodies were detectable in seven donors; one donor presented parallel positivities of HEV-Ag and anti-HEV IgM. The evaluated NAT methods present powerful tools providing sensitive HEV detection. Application of HEV-Ag or anti-HEV IgM screening is currently inferior for the early detection of HEV infection due to the decreased sensitivity compared to NAT methods.

  18. Comparison of real-time SYBR green dengue assay with real-time taqman RT-PCR dengue assay and the conventional nested PCR for diagnosis of primary and secondary dengue infection

    OpenAIRE

    Damodar Paudel; Richard Jarman; Kriengsak Limkittikul; Chonticha Klungthong; Supat Chamnanchanunt; Ananda Nisalak; Robert Gibbons; Watcharee Chokejindachai

    2011-01-01

    Background : Dengue fever and dengue hemorrhagic fever are caused by dengue virus. Dengue infection remains a burning problem of many countries. To diagnose acute dengue in the early phase we improve the low cost, rapid SYBR green real time assay and compared the sensitivity and specificity with real time Taqman® assay and conventional nested PCR assay. Aims: To develop low cost, rapid and reliable real time SYBR green diagnostic dengue assay and compare with Taqman real-time assay and conven...

  19. Comparison of the Diagnostic Value Between Real-Time Reverse Transcription-Polymerase Chain Reaction Assay and Histopathologic Examination in Sentinel Lymph Nodes for Patients With Gastric Carcinoma.

    Science.gov (United States)

    Kwak, Yoonjin; Nam, Soo Kyung; Shin, Eun; Ahn, Sang-Hoon; Lee, Hee Eun; Park, Do Joong; Kim, Woo Ho; Kim, Hyung-Ho; Lee, Hye Seung

    2016-05-01

    Sentinel lymph node (SLN)-based diagnosis in gastric cancers has shown varied sensitivities and false-negative rates in several studies. Application of the reverse transcription-polymerase chain reaction (RT-PCR) in SLN diagnosis has recently been proposed. A total of 155 SLNs from 65 patients with cT1-2, N0 gastric cancer were examined. The histopathologic results were compared with results obtained by real-time RT-PCR for detecting molecular RNA (mRNA) of cytokeratin (CK)19, carcinoembryonic antigen (CEA), and CK20. The sensitivity and specificity of the multiple marker RT-PCR assay standardized against the results of the postoperative histological examination were 0.778 (95% confidence interval [CI], 0.577-0.914) and 0.781 (95% CI, 0.700-0.850), respectively. In comparison, the sensitivity and specificity of intraoperative diagnosis were 0.819 (95% CI, 0.619-0.937) and 1.000 (95% CI, 0.972-1.000), respectively. The positive predictive value of the multiple-marker RT-PCR assay was 0.355 (95% CI, 0.192-0.546) for predicting non-SLN metastasis, which was lower than that of intraoperative diagnosis (0.813, 95% CI, 0.544-0.960). The real-time RT-PCR assay could detect SLN metastasis in gastric cancer. However, the predictive value of the real-time RT-PCR assay was lower than that of precise histopathologic examination and did not outweigh that of our intraoperative SLN diagnosis. © American Society for Clinical Pathology, 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  20. Evaluation of DNA Recombinant Methodologies for the Diagnosis of Plasmodium falciparum and their Comparison with the Microscopy Assay

    Directory of Open Access Journals (Sweden)

    L Urdaneta

    1998-09-01

    Full Text Available Since 1984, DNA tests based on the highly repeated subtelomeric sequences of Plasmodium falciparum (rep 20 have been frequently used in malaria diagnosis. Rep 20 is very specific for this parasite, and is made of 21 bp units, organized in repeated blocks with direct and inverted orientation. Based in this particular organization, we selected a unique consensus oligonucleotide (pf-21 to drive a PCR reaction coupled to hybridization to non-radioactive labeled probes. The pf-21 unique oligo PCR (pf-21-I assay produced DNA amplification fingerprints when was applied on purified P. falciparum DNA samples (Brazil and Colombia, as well as in patient's blood samples from a large area of Venezuela. The performance of the Pf-21-I assay was compared against Giemsa stained thick blood smears from samples collected at a malaria endemic area of the Bolívar State, Venezuela, at the field station of Malariología in Tumeremo. Coupled to non-radioactive hybridization the pf-21-I performed better than the traditional microscopic method with a r=1.7:1. In the case of mixed infections the r value of P. falciparum detection increased to 2.5:1. The increased diagnostic sensitivity of the test produced with this homologous oligonucleotide could provide an alternative to the epidemiological diagnosis of P. falciparum being currently used in Venezuela endemic areas, where low parasitemia levels and asymptomatic malaria are frequent. In addition, the DNA fingerprint could be tested in molecular population studies

  1. Antidepressant utilization after hospitalization with depression: a comparison between non-Western immigrants and Danish-born residents

    DEFF Research Database (Denmark)

    Wallach-Kildemoes, Helle; Thomsen, Louise Thirstrup; Kriegbaum, Margit

    2014-01-01

    BACKGROUND: Antidepressant (AD) therapy is recommended for patients 4-12 months after remission from depression. The aim was to examine whether immigrants (refugees or family reunited immigrants) from non-Western countries are at greater risk than Danish-born residents of 1) not initiating AD the...... income had only minor impact on these associations.\

  2. A Vertical and Horizontal Comparison of Sustainable Development Status in Western China from the Perspective of Ecological Footprint

    Institute of Scientific and Technical Information of China (English)

    2011-01-01

    On the basis of ecological footprint theory, the ecological footprint and ecological carrying capacity of western China and 12 provinces and cities in 2009 are calculated. From the perspective of ecological footprint, the sustainable development status of west China and 12 provinces and cities in 2009 and 1999 are compared vertically and horizontally. Results assume that no matter weighting by using local or national ecological carrying capacity, the west area laid in the unsustainable development state in 2009; in 1999, the west area was on the unsustainable development stage taking the local and national level as the reference and it was at the sustainable stage at the global level. In 2009, most provinces in western China laid on the unsustainable stage at the local, national and global level. Compared with that in 2009,few provinces in western China were on the unsustainable stage at different levels. In general, the sustainable developmental momentum of western China in 2009 was weaker than that in ten years ago and many provinces and cities decreased to unsustainable state.

  3. Challenging the Western Approach to Cultural Comparisons: Young Pupils' Affective Structures Regarding Mathematics in Finland and Chile

    Science.gov (United States)

    Tuohilampi, Laura; Hannula, Markku S.; Varas, Leonor; Giaconi, Valentina; Laine, Anu; Näveri, Liisa; i Nevado, Laia Saló

    2015-01-01

    Large-scale studies measure mathematics-related affect using questionnaires developed by researchers in primarily English-based countries and according to Western-based theories. Influential comparative conclusions about different cultures and countries are drawn based on such measurements. However, there are certain premises involved in these…

  4. Evaluation and comparison of radical scavenging properties of solvent extracts from Justicia adhatoda leaf using DPPH assay.

    Science.gov (United States)

    Jha, Deepak Kumar; Panda, Likun; Ramaiah, Sudha; Anbarasu, Anand

    2014-12-01

    2,2-Diphenyl-1-picrylhydrazyl (DPPH) method is routinely practiced for the assessment of antioxidant activity of compounds and their mixtures. The method is based on the spectrophotometric measurement of DPPH(·) concentration that changes resulting from the DPPH radical reaction with an antioxidant. The amount of remaining DPPH(·) in the examined system is a measure of the antioxidant activity of compounds. Our study aims at exploring the antioxidant properties of Justicia adhatoda leaf extract and comparing the results in terms of effective concentration which scavenges 50 % radical (EC50). The correlation of the activities for both cold and Soxhlet methanolic extracts is reported with DPPH assay. The antioxidant capacity of the methanolic extract derived by two different methods is positively correlated. Correlation between antioxidant capacity and phenolic content of methanolic extract in both the cases indicates the efficiency of the extraction procedure. Positive correlation and p value <0.05 validate the efficiency of the procedures and results.

  5. A rapid and simple assay for lamotrigine in serum/plasma by HPLC, and comparison with an immunoassay.

    Science.gov (United States)

    Morgan, Phillip E; Fisher, Danielle S; Evers, Richard; Flanagan, Robert J

    2011-07-01

    Monitoring serum/plasma concentrations of lamotrigine may be useful under certain circumstances. An HPLC column packed with strong cation-exchange (SCX)-modified microparticulate silica together with a 100% methanol eluent containing an ionic modifier permits direct injection of sample extracts. An HPLC-UV method developed using this principle for the measurement of serum/plasma lamotrigine is simple, sensitive and selective. The analysis time is less than 5 min. Intra- and inter-assay precision and accuracy meet acceptance criteria, and sample stability, and potential interferences from other compounds have been evaluated. There was good agreement with consensus mean results from external quality assessment samples (n = 32). Analysis of patient samples (n = 115) using the HPLC method and the Seradyn QMS® Lamotrigine immunoassay showed that the immunoassay over-estimated lamotrigine by 21% on average.

  6. Comparison of two commercial broadrange PCR and sequencing assays for identification of bacteria in culture-negative clinical samples

    DEFF Research Database (Denmark)

    Stavnsbjerg, Camilla; Frimodt-Moller, Niels; Moser, Claus Ernst

    2017-01-01

    Background Culturing has long been the gold standard for detecting aetiologic agents in bacterial infections. In some cases, however, culturing fails to detect the infection. To further investigate culture-negative samples, amplification and subsequent sequencing of the 16S rRNA gene is often...... applied. The aim of the present study was to compare the current method used at our Department of Clinical Microbiology, based on the MicroSeq ID system (Applied Biosystems, USA) with the Universal Microbe Detection (UMD) SelectNA kit (Molzym, Germany). Methods 76 culture-negative samples were first...... in a real-time PCR and sequenced. Results 22 of 76 samples (28.9%) were positive for bacteria with the UMD SelectNA, which was significantly more (p = 0.0055) than the MicroSeq ID where 11 of 76 samples (14.5%) were positive. The UMD SelectNA assay identified more relevant bacterial pathogens than the Micro...

  7. Comparison of the antiinflammatory effects of Drosera rotundifolia and Drosera madagascariensis in the HET-CAM assay.

    Science.gov (United States)

    Paper, Dietrich H; Karall, Elisabeth; Kremser, Michaela; Krenn, Liselotte

    2005-04-01

    The antiinflammatory effects of ethanol and aqueous extracts from Drosera rotundifolia and from Drosera madagascariensis were compared in vivo in the HET-CAM assay. Both extracts from D. rotundifolia and the ethanol extract from D. madagascariensis showed remarkable efficacy at doses of 500 microg/pellet. The inhibition of the inflammation by the extracts was stronger than that by 50 microg hydrocortisone/pellet. In contrast, there was only a very weak effect observed at a dose of 500 microg/pellet of the water extract from D. madagascariensis. The chemical analyses of the extracts showed that the effect cannot be attributed to naphthoquinones, but might be due to flavonoids. Ellagic acid obviously plays an important role in the antiangiogenic effect of the Drosera extracts. (c) 2005 John Wiley & Sons, Ltd.

  8. A comparison of a solid phase IRC assay and the PSIFT for detection of antibodies to platelets.

    Science.gov (United States)

    Häcker-Shahin, B; Giannitsis, D J

    1992-01-01

    A rapid solid phase indicator red cells assay (IRCA) for detection of platelet antibodies was developed and its sensitivity compared with PSIFT. Platelets were attached to the surface of polystyrene microtitre plate wells by means of a sodium carbonate buffer and centrifugation. Uncovered areas were blocked by a gelatin blocking buffer. After serum incubation bound platelet-specific antibodies were made visible by anti-IgG-coated indicator red cells and a brief centrifugation. A positive result, meaning the presence of an anti-platelet antibody was indicated by red cell adherence over the reaction surface. In the absence of serum antibodies to platelets the indicator red cells formed a pellet. The IRCA showed a high sensitivity; the anti-platelet antibody Thrombocyte was detectable until a dilution of 1:1,600 whereas the same antibody in the PSIFT could only be detected until a dilution of 1:400.

  9. Comparison of murine fibroblast cell response to fluor-hydroxyapatite composite, fluorapatite and hydroxyapatite by eluate assay.

    Science.gov (United States)

    Jantová, Sona; Letasiová, Silvia; Theiszová, Marica; Palou, M

    2009-03-01

    Fluorapatite (FA) is one of the inorganic constituents of bone or teeth used for hard tissue repairs and replacements. Fluor-hydroxyapatite (FHA) is a new synthetic composite that contains the same molecular concentration of OH(-) groups and F(-) ions. The aim of this experiment was to evaluate the cellular responses of murine fibroblast NIH-3T3 cells in vitro to solid solutions of FHA and FA and to compare them with the effect of hydroxyapatite (HA). We studied 24, 48 and 72 h effects of biomaterials on cell morphology, proliferation and cell cycle of NIH-3T3 cells by eluate assay. Furthermore, we examined the ability of FHA, FA and HA to induce cell death and DNA damage. Our cytotoxic/antiproliferative studies indicated that any of tested biomaterials did not cause the total inhibition of cell division. Biomaterials induced different antiproliferative effects increasing in the order HA < FHA < FA which were time- and concentration-dependent. None of the tested biomaterials induced necrotic/apoptotic death of NIH-3T3 cells. On the other hand, after 72 h we found that FHA and FA induced G0/G1 arrest of NIH-3T3 cells, while HA did not affect any cell cycle phases. Comet assay showed that while HA demonstrated weaker genotoxicity, DNA damage induced by FHA and FA caused G0/G1 arrest of NIH-3T3 cells. Fluoridation of hydroxyapatite and different FHA and FA structure caused different cell response of NIH-3T3 cells to biomaterials.

  10. Comparison of invasiveness among surface-adherent variants of Listeria monocytogenes in Caco-2 cell culture assays.

    Science.gov (United States)

    Kushwaha, Kalpana; Muriana, Peter M

    2010-03-31

    The persistence of Listeria monocytogenes in meat processing plants is believed to be partly linked with their adherence properties to abiotic surfaces that may allow contamination of ready-to-eat (RTE) foods. Isolates of raw meats, RTE meats and environmental samples from RTE meat processing facilities that demonstrated differences in adherence to abiotic surfaces (glass, plastic, stainless steel, and rubber) were tested for cellular adherence and invasion in the Caco-2 human cell line. Strains of L. monocytogenes were classified into strong (CW50, 99-38, CW77, SM5 and CW62) and weakly adherent (CW34, CW35, CW72, SM3, J7 and J126) based on a microplate adherence assay using the fluorescing substrate, 5,6-CFDA. These strains were tested for adherence and invasion in cell culture assays with Caco-2 cells. At long incubation time (2h) and high multiplicity of infection (MOI, 100:1) we observed equivalent cellular adherence and invasiveness. After optimizing conditions for time of infection (15-, 30-, 60-, 90-, and 120-min) and MOI (100:1, 10:1, 1:1, and 0.1:1), we found that under the conditions of 15-min infection time at an MOI of 10:1 (Listeria:Caco-2 cells), we again observed equivalent cellular adherence by the two distinct surface-adherent groups, but we only recovered invading Listeria from the group that was strongly adherent to environmental surfaces. The data indicate that a surface factor that provides for strong environmental surface adherence may also be involved with internal cellular virulence and survival and implicates strongly adherent strains as possibly being of greater invasiveness than weakly adherent strains.

  11. Inter-laboratory comparison to validate the dicentric assay as a cytogenetic triage tool for medical management of radiation accidents

    Energy Technology Data Exchange (ETDEWEB)

    Beinke, Christina, E-mail: christinabeinke@bundeswehr.org [Bundeswehr Institute of Radiobiology Affiliated to the University of Ulm, Neuherbergstrasse 11, 80937 Munich (Germany); Oestreicher, Ursula [Federal Office for Radiation Protection, Neuherberg (Germany); Riecke, Armin [Department for Internal Medicine, Federal Armed Forces Hospital, Ulm (Germany); Kulka, Ulrike [Federal Office for Radiation Protection, Neuherberg (Germany); Meineke, Viktor [Bundeswehr Institute of Radiobiology Affiliated to the University of Ulm, Neuherbergstrasse 11, 80937 Munich (Germany); Romm, Horst [Federal Office for Radiation Protection, Neuherberg (Germany)

    2011-09-15

    Radiation accidents with exposure of human beings can assume huge dimensions concerning occurring health impairments and essential medical resources such as personnel, patient care management and appropriate medical facilities. Particularly in mass-casualty events, a rapid sorting and allocation of victims to treatment is needed and their classification in medical treatment groups has to be conducted as fast as possible. For triage purposes several approaches can be considered. Clinical signs and symptoms are extremely helpful in estimating radiation effects on an organ-based level, whereas the assessment of radiation effects based on cytogenetic biodosimetry tools is the alternative approach. For both systems there are pros and cons with respect to the usefulness for specific applications, such as individual cases versus mass-casualty screening or whole- versus partial-body exposures. Among the biodosimetry tools the dicentric chromosome assay (DCA) is considered as the 'gold standard' for biodosimetry after an acute radiation exposure. Recently, steady progress in standardization and harmonization of the DCA has occurred, in order to enable the validated performance of the DCA in the frame of cooperative response of biodosimetry networks during a large scale radiological scenario. Using the DCA in triage mode which allows the stratification of radiation exposed victims into broad 1.0 Gy categories only 20-50 metaphase cells per subject are scored instead of the 500-1000 scored for routine analysis. Our data show that there are significant differences between the dicentric yields after 1.0 Gy and 3.0 Gy {gamma}-ray ex vivo exposure of blood suggesting this assay as suitable for the distinction between high and low dosed exposed individuals. These preliminary findings indicate the usefulness of the DCA also for therapeutic decision making.

  12. Comparison of four functionalization methods of gold nanoparticles for enhancing the enzyme-linked immunosorbent assay (ELISA)

    Science.gov (United States)

    Ciaurriz, Paula; Fernández, Fátima; Tellechea, Edurne; Moran, Jose F

    2017-01-01

    The enzyme-linked immunosorbent assay (ELISA) technique is based on the specific recognition ability of the molecular structure of an antigen (epitope) by an antibody and is likely the most important diagnostic technique used today in bioscience. With this methodology, it is possible to diagnose illness, allergies, alimentary fraud, and even to detect small molecules such as toxins, pesticides, heavy metals, etc. For this reason, any procedures that improve the detection limit, sensitivity or reduce the analysis time could have an important impact in several fields. In this respect, many methods have been developed for improving the technique, ranging from fluorescence substrates to methods for increasing the number of enzyme molecules involved in the detection such as the biotin–streptavidin method. In this context, nanotechnology has offered a significant number of proposed solutions, mainly based on the functionalization of nanoparticles from gold to carbon which could be used as antibody carriers as well as reporter enzymes like peroxidase. However, few works have focused on the study of best practices for nanoparticle functionalization for ELISA enhancement. In this work, we use 20 nm gold nanoparticles (AuNPs) as a vehicle for secondary antibodies and peroxidase (HRP). The design of experiments technique (DOE) and four different methods for biomolecule loading were compared using a rabbit IgG/goat anti-rabbit IgG ELISA model (adsorption, directional, covalent and a combination thereof). As a result, AuNP probes prepared by direct adsorption were the most effective method. AuNPs probes were then used to detect gliadin, one of the main components of wheat gluten, the protein composite that causes celiac disease. With this optimized approach, our data showed a sensitivity increase of at least five times and a lower detection limit with respect to a standard ELISA of at least three times. Additionally, the assay time was remarkably decreased. PMID:28243563

  13. Safety analysis of FOLFOX4 treatment in colorectal cancer patients: a comparison between two Asian studies and four Western studies.

    Science.gov (United States)

    Sugihara, Kenichi; Ohtsu, Atsushi; Shimada, Yasuhiro; Mizunuma, Nobuyuki; Lee, Po-Huang; de Gramont, Aimery; Goldberg, Richard M; Rothenberg, Mace L; André, Thierry; Brienza, Silvano; Gomi, Katsushige

    2012-06-01

    Oxaliplatin-based therapy, notably FOLFOX4 (5-fluorouracil, leucovorin, and oxaliplatin), is a standard regimen approved globally for the treatment of metastatic colorectal cancer, and as adjuvant treatment of colon cancer. As part of the Japanese submission for the adjuvant indication, the safety profile of FOLFOX4 regimen was compared in Asian and Western patients. A total of 3359 patients with colorectal cancer treated with the FOLFOX4 regimen were included in the analyses: 1515 from 2 Asian studies (Japanese Post Marketing Surveillance and Multicenter Asia Study in Adjuvant Treatment of Colon Cancer with Oxaliplatin/5-FU/LV), and 1844 from 4 Western studies (EFC2962, N9741, EFC4584, and Multicenter International Study of Oxaliplatin/5-Fluorouracil/Leucovorin in the Adjuvant Treatment of Colon Cancer). Doses administered and safety parameters were analyzed by using common definitions and programs. Demographic and baseline characteristics were comparable between Asian and Western patients. Patients received FOLFOX4 for a median of 6-12 cycles, which ranged from 16 to 28 weeks. Median dose intensities of oxaliplatin and of 5-fluorouracil, bolus and infusion, were within the ranges of 33 to 36 mg/m(2)/week, 297 to 338 mg/m(2)/week, and 467 to 510 mg/m(2)/week, respectively. Most frequently reported adverse events (AE) included hematologic, gastrointestinal, and neurosensory adverse events (NSAE). The incidence of grade ≥3 neutropenia ranged from 37% (422 of 1134) to 52% (83 of 159) in Asian and 41% (455 of 1108) to 56% (144 of 259) in Western studies; of diarrhea, ranged from 1.4% (3 of 222) to 6.3% (10 of 159) and 11% (30 of 268 or 120 of 1108) to 14% (36 of 259); of NSAEs, from 1.9% (21 of 1134) to 4.4% (7 of 159) and 9.3% (25 of 268) to 19% (39 of 209); and of allergic reactions, from 0.6% (7 of 1134) to 3.1% (5 of 159) and 1.1% (3 of 268) to 3.0% (33 of 1108), respectively. The probability of grade ≥3 NSAEs and diarrhea was statistically significantly lower

  14. Comparison of air-sea fluxes of CO2 in the Southern Ocean and the western Arctic Ocean

    Institute of Scientific and Technical Information of China (English)

    CHEN Liqi; GAO Zhongyong; YANG Xulin; WANG Weiqiang

    2004-01-01

    The data were collected during Chinese Arctic and Antarctic Expeditions in the western Arctic Ocean and the marginal sea ice zone (MSIZ) of the Southern Ocean, respectively in the boreal summer from July to September of 1999 and in the austral summer from December of 1999 to January of 2000. The concentrations of CO2 in surface water of the survey regions would mostly present lower than those in the atmosphere. A significant biological driving force could also been observed in summer waters in both of the above oceans. Air to sea CO2 fluxes were also calculated to compare oceanic uptake capacity of CO2 in both oceans with the world oceans using Liss, Wanninkhof,and Jacobs' s methods. The averaged CO2 fluxes of air to sea in the western Arctic Ocean or in the MSIZ of the Southern Ocean doubled that in the world oceans.

  15. Growth ages of ferromanganese crusts from the western and central Pacific: Comparison between nannofossil analysis and 10Be dating

    Institute of Scientific and Technical Information of China (English)

    CHENG Zhenbo; SHI Fengdeng; M.Hiroyuki; SHI Xuefa; WU Yonghua; SU Xin; LI Xiaoyan; WANG Kunshan; YANG Yongliang; GE Shulan; JU Xiaohua

    2006-01-01

    Based on results of nannofossil analysis and 10Be dating in ferromanganese crusts M1-1 and A1-1 (no nannofossils were found in it), from the western and central Pacific respectively, it is found that the crust growth ages from nannofossil biostratigraphy agree well with those based on 10Be isotope analysis. Both crusts have three growth layers, and the oldest layer was deposited in Miocene at about 12.80 Ma. The maximum, minimum, and average growth rates of crust A1-1 (from the central Pacific)are 8.11, 1.92 and 3.47 mm/Ma, and those of crust M1-1 (from the western Pacific) are 2.93, 0.47, and0.94 mm/Ma.

  16. A Comparison of Typeface Design between Chinese and Western Characters%中西文字体设计比较

    Institute of Scientific and Technical Information of China (English)

    徐雪松; 杨钊

    2014-01-01

    随着中西方文化交流增多和数字化技术的发展,汉字的研究和字体设计开发方兴未艾。由于中西文字发展历史和东西方文化的不同,汉字与拉丁文字在字体造型设计和审美等方面存在诸多差异。文章从分析中西文字体的演进入手,探究中西文字体设计的不同特征和共同点,阐述了现代汉字字体设计的发展趋向,旨在通过对中西文字体设计的系统比较,对汉字字体设计形成有效借鉴,促进汉字设计艺术的创新发展。%With the increasing communication between Chinese and western culture and the development of digital technology, the Chinese character typeface design is growing rapidly. Because of the different history of Chinese character and Latin alphabet, also the differences between eastern and western culture, there are many differences in their typography and aesthetic. From the analysis of the evolution of Chinese and western characters, this paper explored the similarities and differences between typeface design of these two characters, and analyzed the development trend of modern Chinese typeface design. It systematically compared the typeface design characteristics between Chinese and western characters , aiming to give effective reference and promote the innovation and development of Chinese typography.

  17. Particle counting assay for anti-toxoplasma IgG antibodies. Comparison with four automated commercial enzyme-linked immunoassays.

    Science.gov (United States)

    Galanti, L M; Dell'Omo, J; Wanet, B; Guarin, J L; Jamart, J; Garrino, M G; Masson, P L; Cambiaso, C L

    1997-09-24

    An assay for anti-toxoplasma IgG antibodies based on agglutination of latex particles was set up and compared with commercial immunoassays. The reaction was measured by instrumental counting of particles remaining unagglutinated. The running time was 45 min. This test (PaC) was compared using 243 serum samples with four automated commercial immunoassays: the Enzymum test Toxo IgG (ES300, Boehringer), the Vidas Toxo IgG (Biomérieux), the IMX Toxo IgG (Abbott), the Magia Toxoplasma gondii IgG (Merck). The mean values (+/- SD) obtained by IMX (25 IU +/- 68) and ES300 (45 IU +/- 142) were significantly lower than the values obtained by Vidas (73 IU +/- 237, p Magia (80 IU +/- 300, p < 10(-4) and p = 0.0005) and by PaC (70 IU +/- 260, p < 10(-4) and p = 0.0126). The correlations between PaC and Toxo IgG Boehringer, Biomérieux, Abbott, Merck were r = 0.97, r = 0.98, r = 0.94, r = 0.98, respectively. The correlation coefficients between the enzyme-immunoassays ranged from 0.96 to 0.99. All positive samples by PaC were found to be positive by enzyme-immunoassays except for eight sera which were doubtful positives by the Enzymum test ToxoIgG from Boehringer. No negative sample by PaC was found positive by any of the enzyme-immunoassays. In PaC, when two latex preparations coated with different antigen were compared, the correlation was rather weak (r = 0.93) suggesting that the selection of the antigen can be critical. In conclusion, the four automated commercial immunoassays now available gave similar results. However, the discrepancies observed in this study underlined the importance of clinical and biological follow-up of the patients and the necessity to confirm the result. The introduction of a new technique such as PaC, which is now available for a large variety of assays in Clinical Chemistry and Microbiology, is justified by its intrinsic advantage of homogeneity. Therefore, automation is easy as well as the control of possible interference.

  18. British Attitudes Towards Sexuality in Men and Women with Intellectual Disabilities: A Comparison Between White Westerners and South Asians.

    Science.gov (United States)

    Sankhla, Deepak; Theodore, Kate

    Although sexuality is a fundamental aspect of human existence, public attitudes towards the sexuality of people with intellectual disabilities may vary. In particular, different ethnic communities may have different perspectives. These differing perspectives may impact on the opportunities and support available for people with intellectual disabilities to express sexuality within 'normalized' life experiences. Despite the South Asian population being one of the largest minority ethnic groups residing within the UK, few studies have aimed to understand how South Asian attitudes towards the sexuality of people with intellectual disabilities may differ from White Western perspectives. This study used an online questionnaire to investigate public attitudes towards the sexuality of people with intellectual disabilities within a UK sample (n = 331). Attitudes between people from White Western (n = 184) and South Asian backgrounds (n = 147) were compared with the use of five scales measuring attitudes towards sexuality. Whilst overall attitudes appeared to be generally positive, South Asian participants were found to have significantly more negative attitudes towards the sexual control and sexual rights of people with intellectual disabilities compared to White Westerners. These differences remained significant even after factors known to influence such attitudes were taken into consideration. These findings implicate the need to develop culturally sensitive interventions to improve knowledge and awareness of sexual needs of people with intellectual disabilities. This paper discusses these implications further, the limitations of the present study and suggested directions for future research.

  19. [Nursing-home-acquired pneumococcal pneumonia--comparison of sputum cultures with Binax NOW Streptococcus pneumoniae urinary antigen assay].

    Science.gov (United States)

    Rikimaru, Toru; Nishiyama, Mamoru; Yonemitsu, Junko; Nagabuchi, Masako; Shimada, Akiko; Koga, Takeharu; Aizawa, Hisamichi

    2008-11-01

    To clarify the clinical significance of Pneumococcal pneumonia in nursing-home-acquired pneumonia, we examined the positive disease rate of using sputum cultures and the Binax NOW Streptococcus pneumoniae urinary antigen assay in 154 nursing-home patients with pneumonia. These included 54 males and 100 females with a mean age of 86.2 years. Bacteriological findings for sputum culture in 130 patients showed Streptococcus pneumoniae to be cultured in 11 cases (8%). In 72 in whom the Streptococcus pneumoniae-urinary antigen test (Binax NOW) was done, the urinary-antigen-positive rate (26/72 ; 36%) was higher than the culture positive rate for S. pneumoniae. Both examinations were done in 64 patients, among whom 5 in whom S. pneumoniae was cultured also had positive results for the urinary antigen test. Almost half of those undergoing percutaneous endoscopic gastroscopy (PEG) tube nutrition had positive results for the urinary antigen test, but not all such patients had positive cultures for S. pneumoniae. Although the culture-positive rate for S. pneumoniae in sputum was low, we concluded that S. pneumoniae was frequently linked to nursing-home-acquired pneumonia, especially in "total-care" patients.

  20. High-throughput fluorescence screening assay for the identification and comparison of antimicrobial peptides' activity on various yeast species.

    Science.gov (United States)

    Kodedová, Marie; Sychrová, Hana

    2016-09-10

    New antifungal compounds that circumvent the resistance of the pathogen by directly damaging yeast cell surface structures are promising agents for the treatment of fungal infections, due to their different mechanism of action from current clinically used antifungal drugs. We present here a rapid and cost-effective fluorescence method suitable for identifying new potent drugs that directly target yeast cell surface structures, causing cell permeabilization and thus bypassing the multidrug resistance mechanisms of pathogens. The fluorescence assay enabled us to detect with high sensitivity damage to the Candida plasma membrane (its hyperpolarization and permeabilization) as a result of short-term exposure to the antifungal compounds. Results can be obtained in 1-2h with minimal effort and consumption of the tested compounds, also 96 samples can be analysed simultaneously. We used this method to study antimicrobial peptides isolated from the venom of bees and their synthetic analogs, compare the potency of the peptides and determine their minimal effective concentrations. The antimicrobial peptides were able to kill yeast cells at low concentrations within a 15-min treatment, the LL-III peptide exhibited a broad spectrum of antifungal activity on various Saccharomyces, pathogenic Candida and osmotolerant yeast species.

  1. Comparison of agar gel immunodiffusion test, enzyme-linked immunosorbent assay and PCR in diagnostics of enzootic bovine leukosis

    Directory of Open Access Journals (Sweden)

    Malovrh Tadej

    2005-01-01

    Full Text Available Bovine leukaemia virus (BLV is a retrovirus that induces a chronic infection in cattle. Once infected, cattle remain virus carriers for life and start to show an antibody response within a few weeks after infection. Eradication and control of the disease are based on early diagnostics and segregation of the carriers. The choice of a diagnostic method depends on the eradication programme, money resources and characteristics of the herd to be analysed. The agar gel immunodiffusion (AGID test has been the serological test of choice for routine diagnosis of serum samples. Nevertheless, in more recent years, the enzyme-linked immunosorbent assay (ELISA has replaced the AGID for large scale testing. For this purpose, commercially available BLV-ELISA kits were compared to the AGID and to the polymerase chain reaction (PCR method performed with two sets of primers, amplifying env region. The ELISA kit based on the p24 core protein was found to be less specific and served as a screening test. The ELISA kit based on the envelope glycoprotein (gpSI served as a verification test and gave a good correlation with the AGID test and PCR method. However, ELISA showed a higher sensitivity than AGID. The p24 based ELiSA was useful for screening a large number of samples, whereas gp51 based ELISA, AGID and PCR were more important for detecting the antibody response against the individual BLV-proteins and therefore for verification of the infection with BLV.

  2. Comparison of enzyme-linked immunosorbent assay and radioimmunoassay for prostate-specific acid phosphatase in prostatic disease

    Energy Technology Data Exchange (ETDEWEB)

    Griffiths, J.; Rippe, D.F.; Panfili, P.R.

    1982-01-01

    Results of an enzyme-linked immunosorbent assay (ELISA) are compared with those of a standard radioimmunoassay (RIA) for detection and quantitation of prostate-specific acid phosphatase (EC 3.1.3.2) in serum. Control subjects, patients with benign prostatic hyperplasia, and patients in all four clinical stages of prostatic adenocarcinoma were tested. The upper limit of normal (95%of the population) by the ELISA was 2.0 ..mu..g/L, and by the RIA was 2.2 ..mu..g/L. In prostatic a denocarcinoma stage I (not detectable by digital rectal examination), ELISA was slightly more sensitive than RIA, but sensitivity was still relatively low (20%). As tumor mass increased (stages II through IV), the frequency of increased concentrations of prostatic acid phosphatase in serum also increased. We confirmed this increase in circulating enzyme in some cases of benign prostatic hyperplasia and suggest that this finding is related to either acinar cytolysis or an increase in acini size and number. Although prostate-specific acid phosphatase is not a cancer-specific enzyme, we conclude that its measurement may be of considerable value in monitoring prostatic disease.

  3. Bluetongue in Bosnia: comparisons of competitive enzyme-linked immunosorbent assay and standard agar gel immunodiffusion tests.

    Science.gov (United States)

    Velić, L; Velić, R; Bajrović, T; Dukić, B; Camo, D

    2004-01-01

    At the end of August 2002, clinical symptoms of bluetongue (BT) (fever between 39 degrees C and 41 degrees C, muco-purulent or bloody nasal discharge, oedema of the lips and the intramandibular space, foot lesions including laminitis and coronitis in some cases, diarrhoea and dysentery) were recorded in Pramenka sheep flocks in north-east Bosnia in August 2002. A total of 9 599 serum samples (ovine: 8 967; bovine: 632) from 40 communities of Bosnia and Herzegovina were tested for the presence of anti-bluetongue virus (BTV) antibodies using competitive enzyme-linked immunosorbent assay (c-ELISA) and the standard agar gel immunodiffusion (AGID) test. The c-ELISA revealed BTV-seropositive reactions in 187 (1.94%) samples and the AGID test detected 141 (1.53%) cases. Complete agreement was recorded between the c-ELISA and AGID test results for bovine sera. These results indicate that the ability of c-ELISA to detect anti-bluetongue virus antibodies in ovine sera was superior to that of the AGID. All positive sera were collected from animals in the river areas of Bosnia and Herzegovina.

  4. Comparison of enzyme-linked immunosorbent assays and virus neutralization test for detection of antibodies to avian pneumovirus.

    Science.gov (United States)

    Alkahalaf, A N; Halvorson, D A; Saif, Y M

    2002-01-01

    Two different whole-virus enzyme-linked immunosorbent assays (ELISAs), developed in Ohio (OH) with APV/Minnesota/turkey/2a/97 and in Minnesota (MN) with APV/Colorado/turkey/97, and the virus neutralization (VN) test were used to test 270 turkey serum samples from 27 Minnesota turkey flocks for avian pneumovirus (APV) antibodies. In addition, 77 turkey serum samples and 128 ostrich serum samples from Ohio were tested. None of the turkey samples from Ohio had antibodies to APV by the VN test and OH ELISA. The ostrich samples were only tested with the VN test and were all negative for antibodies to APV. For the Minnesota serum samples, 107, 115, and 120 were positive by the VN test, the OH ELISA, and the MN ELISA, respectively. The Kappa values of 0.938 and 0.825 showed excellent agreement between the VN test and the OH ELISA and the MN ELISA, respectively, for detection of antibodies to the APV. The OH ELISA and MN ELISA had sensitivities of 1.0 and 0.953, specificities of 0.950 and 0.889, and accuracies of 0.970 and 0.914, respectively. Our results indicate that the 3 methods are sensitive and specific for diagnosis of the APV infection.

  5. Non-invasive monitoring of physiological stress in the Western lowland gorilla (Gorilla gorilla gorilla): validation of a fecal glucocorticoid assay and methods for practical application in the field.

    Science.gov (United States)

    Shutt, Kathryn; Setchell, Joanna M; Heistermann, Michael

    2012-11-01

    Enzymeimmunoassays (EIAs) allow researchers to monitor stress hormone output via measurement of fecal glucocorticoid metabolites (FGCMs) in many vertebrates. They can be powerful tools which allow the acquisition of otherwise unobtainable physiological information from both captive animals and wild animals in remote forest habitats, such as great apes. However, methods for hormone measurement, extraction and preservation need to be adapted and validated for field settings. In preparation for a field study of Western lowland gorillas (Gorilla gorilla gorilla) in the Central African Republic we used samples from captive gorillas collected around opportunistic stressful situations to test whether four different glucocorticoid EIAs reflected adrenocortical activity reliably and to establish the lag-time from the stressor to peak excretion. We also validated a field extraction technique and established a simple, non-freezer-reliant method to preserve FGCMs in extracts long-term. We determined the rate of FGCM change over 28 days when samples cannot be extracted immediately and over 12h when feces cannot be preserved immediately in alcohol. Finally, we used repeat samples from identified individuals to test for diurnal variation in FGCM output. Two group-specific assays measuring major cortisol metabolites detected the predicted FGCM response to the stressor reliably, whereas more specific cortisol and corticosterone assays were distinctly less responsive and thus less useful. We detected a lag time of 2-3 days from stressor to peak FGCM excretion. Our field extraction method performed as well as an established laboratory extraction method and FGCMs in dried extracts stored at ambient temperatures were as stable as those at -20 °C over 1 yr. Hormones in non-extracted feces in alcohol were stable up to 28 days at ambient temperatures. FGCMs in un-fixed gorilla feces deteriorated to almost 50% of the original values within 6h under field conditions. We detected no diurnal

  6. Comparison of indirect immunofluorescence assays for diagnosis of scrub typhus and murine typhus using venous blood and finger prick filter paper blood spots.

    Science.gov (United States)

    Phetsouvanh, Rattanaphone; Blacksell, Stuart D; Jenjaroen, Kemajittra; Day, Nicholas P J; Newton, Paul N

    2009-05-01

    We performed indirect immunofluorescence assays (IFAs) to compare levels of IgM and IgG antibodies to Orientia tsutsugamushi and Rickettsia typhi in admission-phase serum samples and filter paper blood spots (assayed immediately and stored at 5.4 degrees C and 29 degrees C for 30 days) collected on the same day from 53 adults with suspected scrub typhus and murine typhus admitted to Mahosot Hospital Vientiane, Lao People's Democratic Republic. The sensitivities and specificities of admission-phase filter paper blood spots in comparison to paired sera were between 91% and 95% and 87% and 100%, respectively, for the diagnosis of scrub typhus and murine typhus. The classification of patients as having or not having typhus did not significantly differ after storage of the blood spots for 30 days (P > 0.4) at 5.4 degrees C and 29 degrees C. Because filter paper blood samples do not require sophisticated and expensive storage and transport, they may be an appropriate specimen collection technique for the diagnosis of rickettsial disease in the rural tropics.

  7. Enzyme-linked immunosorbent assay for group A Streptococcal anti-DNase B in human sera, using recombinant proteins - Comparison to the DNA methyl green micromethod.

    Science.gov (United States)

    Das, Sarita; Dileepan, T; Johnson, D R; Kaplan, E L; Patrick Cleary, P

    2017-09-19

    Among the four known Streptococcal nucleases comprising of DNase A, B, C and D; DNase B is the most common, and determination of the levels of antibody to DNase B (ADB) is often used to confirm a clinical diagnosis of Streptococcus pyogenes/group A Streptococcal (GAS) infection. The commonly used assays for antibodies that neutralize DNase B or streptolysin O activity use partially purified antigens that often fail to detect antibody changes subsequent to culture documented infections. Therefore, an enzyme-linked immunosorbent assay (ELISA) was developed employing his-tagged recombinant DNase B as plate antigen for comparison to the commonly used DNA methyl green micromethod (DMGM). DNAs from various Streptococcal species were screened for presence of dnaseB gene by PCR. Measurements of ADB in sera collected from subjects belonging to different ages, and ethnic groups were used to compare the two methods. dnaseB was not detected by PCR in DNA samples isolated from different strains of group B (GBS), C (GCS) and G (GGS) Streptococci. The ADB based ELISA proved to be highly sensitive and more responsive to changes in antibody concentration than DMGM. Use of recombinant DNase B eliminates the variability associated with the enzyme, partially purified from Streptococcal culture supernatants from various commercial sources and may provide a more reliable source of antigen to a wider group of laboratories concerned with GAS diagnosis. Copyright © 2017. Published by Elsevier B.V.

  8. Analysis of chromameter results obtained from corticosteroid-induced skin blanching assay: comparison of visual and chromameter data.

    Science.gov (United States)

    Schwarb, F P; Smith, E W; Haigh, J M; Surber, C

    1999-05-01

    In a Guidance document, the American FDA recommends the use of a Minolta chromameter rather than the human eye for the quantitative assessment of the pharmacodynamic blanching response produced by topical application of corticosteroids. The purpose of this study was to compare the appropriateness of the human eye and two models of chromameter for the estimation of skin blanching, in terms of the quality of the data generated by each method. The corticosteroid-induced skin blanching from four different betamethasone 17-valerate cream formulations was compared in a typical human skin blanching trial. The optimized assay methodology routinely practised in our laboratories was utilized. The blanching responses were assessed visually by three trained, independent observers and recorded by two chromameters (Minolta model CR-200 and model CR-300). The topical availability of the four creams was determined using visual scoring and chromameter measurements. All data were manipulated in such a manner as to produce a blanching response versus time profile from which AUBC analysis could be performed. Good correlation was observed between the visual assessments made by three independent observers. In contrast, moderate correlation was determined between visual, CR-200 and CR-300 measurements. Surprisingly, no direct linear relationship between the AUBCs produced by the two chromameters was observed indicating that the quality of the data obtained from the two instruments may not be equal. This investigation also indicated that the use of the chromameter is not completely objective. Visual scoring and chromameter measurement produce data sets that differ in quality. Each procedure needs to be validated and investigators have to be trained for both visual assessment and the operation of the chromameter, particularly with regard to the manipulation of the measuring head of the instrument.

  9. Comparison of seven commercial enzyme-linked immunosorbent assays for the detection of anti-diphtheria toxin antibodies.

    Science.gov (United States)

    Zasada, A A; Rastawicki, W; Śmietańska, K; Rokosz, N; Jagielski, M

    2013-07-01

    Determination of immune status of patients to diphtheria toxin is based mainly on the results of commercially available ELISA kits. The aim of the present study was to compare the results obtained by ELISAs from seven different manufacturers: Mikrogen, Immunolab, Sekisui Virotech, NovaTec, Virion\\Serion, IBL International and Euroimmun. All assays were performed according to the manufacturers' instructions. The concentrations of the anti-diphtheria toxin antibodies in 72 serum samples were calculated on the basis of curves constructed from standards supplied by manufacturers and the new reference material-International Standard for Diphtheria Antitoxin (10/262). The repeatability and reproducibility of all the ELISA kits tested were good. Number of sera with concentrations of the anti-diphtheria toxin antibodies below the WHO-recommended level of protection (0.1 IU/ml) were dependent on the ELISA used: Mikrogen, 20/72 samples (27.7%); Immunolab, 11/72 samples (15.3%); Sekisui Virotech, 0/72 samples (0%); NovaTec 18/72 samples (25.0%); Serion 12/72 samples (16.7%); IBL International, 7/72 samples (9.7 %); and Euroimmun, 17/72 samples (23.6%). However, the results obtained in particular ELISAs, with the exception of Sekisui Virotech, were much more consistent when the concentrations of the anti-diphtheria toxin antibodies in 72 sera measured by using curves constructed from the International Standard 10/262. The data obtained clearly demonstrated that manufacturer-dependent differences between anti-diphtheria IgG ELISA kits exist. The differences in recommendations accepted by the individual manufacturers together with differences shown in our studies in sensitivity greatly affect the clinical interpretation of results.

  10. Comparison of Two MicroRNA Quantification Methods for Assaying MicroRNA Expression Proifles in Wheat (Triticum aestivum L.)

    Institute of Scientific and Technical Information of China (English)

    HAN Ran; YAN Yan; ZHOU Peng; ZHAO Hui-xian

    2014-01-01

    Two microRNA (miRNA) quantification methods, namely, poly(A) reverse transcription (RT)-quantitative real-time polymerase chain reaction (qPCR) and stem-loop RT-qPCR, have been developed for quantifying miRNA expression. In the present study, ifve miRNAs, including miR166, miR167, miR168, miR159, and miR396, with different sequence frequencies, were selected as targets to compare their expression proifles in ifve wheat tissues by applying the two methods and deep sequencing. The study aimed to determine a simple, reliable and high-throughput method for detecting miRNA expressions in wheat tissues. Results showed that the miRNA expression proifles determined by poly(A) RT-qPCR were more consistent with those obtained by deep sequencing. Further analysis indicated that the correlation coefifcients of the data obtained by poly(A) RT-qPCR and deep sequencing (0.739, P 0.01) were higher than those obtained by stem-loop RT-qPCR and deep sequencing (0.535, P 0.01). The protocol used for poly(A) RT-qPCR is simpler than that for stem-loop RT-qPCR. Thus, poly(A) RT-qPCR was a more suitable high-throughput assay for detecting miRNA expression proifles. To the best of our knowledge, this study was the ifrst to compare these two miRNA quantiifcation methods. We also provided useful information for quantifying miRNA in wheat or other plant species.

  11. School based screening for tuberculosis infection in Norway: comparison of positive tuberculin skin test with interferon-gamma release assay

    Science.gov (United States)

    Winje, Brita Askeland; Oftung, Fredrik; Korsvold, Gro Ellen; Mannsåker, Turid; Ly, Ingvild Nesthus; Harstad, Ingunn; Dyrhol-Riise, Anne Margarita; Heldal, Einar

    2008-01-01

    Background In Norway, screening for tuberculosis infection by tuberculin skin test (TST) has been offered for several decades to all children in 9th grade of school, prior to BCG-vaccination. The incidence of tuberculosis in Norway is low and infection with M. tuberculosis is considered rare. QuantiFERON®TB Gold (QFT) is a new and specific blood test for tuberculosis infection. So far, there have been few reports of QFT used in screening of predominantly unexposed, healthy, TST-positive children, including first and second generation immigrants. In order to evaluate the current TST screening and BCG-vaccination programme we aimed to (1) measure the prevalence of QFT positivity among TST positive children identified in the school based screening, and (2) measure the association between demographic and clinical risk factors for tuberculosis infection and QFT positivity. Methods This cross-sectional multi-centre study was conducted during the school year 2005–6 and the TST positive children were recruited from seven public hospitals covering rural and urban areas in Norway. Participation included a QFT test and a questionnaire regarding demographic and clinical risk factors for latent infection. All positive QFT results were confirmed by re-analysis of the same plasma sample. If the confirmatory test was negative the result was reported as non-conclusive and the participant was offered a new test. Results Among 511 TST positive children only 9% (44) had a confirmed positive QFT result. QFT positivity was associated with larger TST induration, origin outside Western countries and known exposure to tuberculosis. Most children (79%) had TST reactions in the range of 6–14 mm; 5% of these were QFT positive. Discrepant results between the tests were common even for TST reactions above 15 mm, as only 22 % had a positive QFT. Conclusion The results support the assumption that factors other than tuberculosis infection are widely contributing to positive TST results in

  12. Airborne Measurements of Western U.S. Wildfire Emissions: Comparison with Prescribed Burning and Air Quality Implications

    Science.gov (United States)

    Liu, Xiaoxi; Huey, L. Gregory; Yokelson, Robert J.; Selimovic, Vanessa; Simpson, Isobel J.; Mueller, Markus; Jimenez, Jose L.; Campuzano-Jost, Pedro; Beyersdorf, Andreas J.; Blake, Donald R.; hide

    2017-01-01

    Wildfires emit significant amounts of pollutants that degrade air quality. Plumes from three wildfires in the western U.S. were measured from aircraft during the Studies of Emissions and Atmospheric Composition, Clouds and Climate Coupling by Regional Surveys (SEAC4RS) and the Biomass Burning Observation Project (BBOP), both in summer 2013. This study reports an extensive set of emission factors (EFs) for over 80 gases and 5 components of submicron particulate matter (PM1) from these temperate wildfires. These include rarely, or never before, measured oxygenated volatile organic compounds and multifunctional organic nitrates. The observed EFs are compared with previous measurements of temperate wildfires, boreal forest fires, and temperate prescribed fires. The wildfires emitted high amounts of PM1 (with organic aerosol (OA) dominating the mass) with an average EF that is more than 2 times the EFs for prescribed fires. The measured EFs were used to estimate the annual wildfire emissions of carbon monoxide, nitrogen oxides, total non methane organic compounds, and PM1 from 11 western U.S. states. The estimated gas emissions are generally comparable with the 2011 National Emissions Inventory (NEI). However, our PM1 emission estimate (1530 +/- 570 Gg/yr) is over 3 times that of the NEI PM2.5 estimate and is also higher than the PM2.5 emitted from all other sources in these states in the NEI. This study indicates that the source of OA from biomass burning in the western states is significantly underestimated. In addition, our results indicate that prescribed burning may be an effective method to reduce fine particle emissions.

  13. Direct Comparison of the Histidine-rich Protein-2 Enzyme-linked Immunosorbent Assay (HRP-2 ELISA) and Malaria SYBR Green I Fluorescence (MSF) Drug Sensitivity Tests in Plasmodium falciparum Reference Clones and Fresh ex vivo Field Isolates from Cambodia

    Science.gov (United States)

    2013-07-12

    RESEARCH Open Access Direct comparison of the histidine -rich protein-2 enzyme-linked immunosorbent assay (HRP-2 ELISA) and malaria SYBR green I...Walsh1, David L Saunders1 and Charlotte A Lanteri1* Abstract Background: Performance of the histidine -rich protein-2 enzyme-linked immunosorbent... histidine -rich protein-2 enzyme-linked immunosorbent assay (HRP-2 ELISA) and malaria SYBR green I fluorescence (MSF) drug sensitivity tests in

  14. Comparison of differences between dicentric assay and translocation analysis for biodosimetry in cultured peripheral blood lymphocytes of Korean individuals

    Energy Technology Data Exchange (ETDEWEB)

    Park, Hyun Jin; Park, Mi Young; Seo, Min Ji; Kwon, Hee Kyung; Lee, Su Jae; Lee, Yun Sil; Ji, Young Hoon; Choi, Soo Yong; Cho, Chul Koo; Kim, Tae Hwan; Kang, Chang Mo [Korea Institute of Radiological and Medical Sciences, Seoul (Korea, Republic of)

    2004-07-01

    Chromosome aberrations are considered to be important indicators of induced DNA damage and genomic instability. For this reason, they constitute the main parameter used to monitor individuals exposed to radiation. Biological dosimetry using the analysis of dicentrics in human lymphocytes is well established, especially in case of acute exposure, when the blood samples are taken within a few weeks. However, dicentric analysis is not an adequate parameter in case of chronic exposure, because these aberrations are unstable with time, and have a limited use for dose assessment of past exposures. In contrast to dicentrics, however, translocations are considered stable in cell division and so the yield should not fall with time. In the present study, using FISH-chromosome painting analysis with the dose-response curve for chromosome aberrations, we monitored the stable and unstable chromosome aberrations of 2 Korean's periperal blood lymphocytes irradiated in vitro with {gamma}-rays from {sup 137}Cs (doses between 0.0 and 2.0 Gy). By using the dose-response curve for chromosome aberration, our aim was to estimate the absorbed doses, and then establish comparison with the results obtained by conventional dicentric analysis, thus taking the opportunity to test the validity of chromosome aberration analysis by FISH painting method for retrospective biodosimetry in Korean individual.

  15. A Comparison of Microscopy and Enzyme Linked Immunosorbent Assay for Diagnosis of Giardia lamblia in Human Faecal Specimens.

    Science.gov (United States)

    Jahan, Noor; Khatoon, Razia; Ahmad, Siraj

    2014-11-01

    Giardia lamblia, a flagellate protozoa, is a common causative agent of parasitic diarrhoeal diseases of humans. Laboratory diagnosis mainly consists of direct microscopic examination of stool specimen for trophozoite and cysts of Giardia. However, due to intermittent faecal excretion of parasite, the case may be miss diagnosed and the patient may continue excreting the parasite and infecting others. Therefore, other mode of diagnosis should be looked for, which overcome the above drawbacks of microscopy used alone for diagnosis. The present study was done to evaluate the efficacy of RIDASCREEN Giardia (ELISA) test in comparison to direct microscopy in the diagnosis of Giardia lamblia in stool specimens from patients with diarrhea and other gastrointestinal symptoms. A total of 1680 patients were included in the study and three faecal specimens were taken from each patient which was divided into two parts. One part was used for direct wet mount examination and second part was used to put ELISA by using RIDASCREEN Giardia test. Out of 1680 stool samples, 380 specimens (22.6%) were found to be positive for Giardia lamblia. Maximum cases were detected by RIDASCREEN Giardia (ELISA) test with sensitivity of 100% and specificity of 91.5%. Maximum cases of giardiasis were detected in children less than 10 y of age (12.8%). RIDASCREEN Giardia test is a rapid and effective method with high sensitivity and specificity and detects Giardia antigens in stool specimens even when the count of parasite is low, thus reducing the chances of missing even the asymptomatic cases.

  16. Radioimmunoassay for somatomedin C: comparison with radioreceptor assay in patients with growth-hormone disorders, hypothyroidism, and renal failure

    Energy Technology Data Exchange (ETDEWEB)

    Baxter, R.C.; Brown, A.S.; Turtle, J.R.

    1982-03-01

    An antiserum (Tr4) was raised in rabbits against a basic somatomedin C-like peptide preparation. Using high-immunoreactivity somatomedin C tracer, we compared the performance of radioimmunoassays in which we used the Tr4 antiserum distributed by the National Pituitary Agency (NPA) with that of the human placental-membrane somatomedin radioreceptor asay (RRA). In their cross reactivity towards various somatomedin-like and unrelated peptides, the two radioimmunoassay methods were almost identical, although NPA antiserum, with about fourfold higher titer than Tr4 antiserum, showed a slightly greater sensitivity for most peptides tested. Radioimmunoassay of acid-ethanol-extracted plasma samples from normal persons and acromegalic, hypopituitary, hypothyroid, and renal-failure patients revealed no analytical differences between the antisera (for 122 samples, r = 0.979 between methods). Somatomedin values for acromegalic and hypopituitary samples showed no overlap with normals. Values for hypothyroid and pre-dialysis renal-failure samples were significantly lower than normal. By comparison, the RRA showed greater cross reactivity towards some somatomedin-like peptides and gave significantly lower values than radioimmunoassay for acromegalic and hypothyroid plasma extracts, and significantly higher values for hypopituitary and renal-failure samples. We conclude that the radioimmunoassay methods clearly are of greater diagnostic value than RRA for clinical somatomedin measurement.

  17. Determination of free insulin-like growth factor-I in human serum: comparison of ultrafiltration and direct immunoradiometric assay.

    Science.gov (United States)

    Frystyk, J; Ivarsen, P; Støving, R K; Dall, R; Bek, T; Hagen, C; Ørskov, H

    2001-04-01

    Two fundamentally different methods are currently used for the determination of free insulin-like growth factor-I (IGF-I): ultrafiltration by centrifugation (UF) and direct immunoradiometric assay (IRMA). The aim was to evaluate a commercial IRMA (DSL, Webster, TX, USA) and to compare it with UF. In the IRMA it is recommended that samples be incubated for 2 h at 5;C. When comparing samples (n = 8) incubated for 1 and 2 h, levels increased by 27 +/- 5% (P< 0.0001). When incubating samples at 22;C instead of 5;C, levels increased by 192 +/- 32% (P< 0.0001). Addition of IGF-binding protein-1 (IGFBP-1) to normal sera (n = 6) dose-dependently decreased ultrafiltered free IGF-I only (P< 0.0007). Similarly, UF was more sensitive than IRMA to addition of IGFBP-2 (P< 0.05). In healthy subjects (n = 35) IRMA yielded 20% higher levels than UF (1.09 +/- 0.09 vs 0.91 +/- 0.12 microg/L; P< 0.0001). IRMA and UF yielded similar results in healthy subjects treated with IGF-I (n = 5) or growth hormone (n = 7) and in acromegalic patients (n = 6) before and after somatostatin analogue treatment. However, marked differences were observed in conditions with elevated IGFBP-1 and -2. In type-1 diabetics (n = 23) ultrafiltered free IGF-I was more reduced than IRMA free IGF-I (38 +/- 9 vs 76 +/- 7% of matched controls (n = 13); P< 0.0001). In patients with chronic renal failure (n = 25), IRMA free IGF-I was identical to control levels (n = 13), whereas ultrafiltered free IGF-I was decreased by 51 +/- 7% (P< 0.0001). Similarly, women with anorexia nervosa (n = 9) studied before and after weight gain showed significant changes in ultrafiltered free IGF-I only (P< 0.03). In conclusion, IRMA was not very robust with respect to variations in sample incubation and this may bias results. IRMA generally yielded higher levels than UF, in accordance with the knowledge that IRMA measures free plus readily dissociable IGF-I. IRMA was less affected than UF by added IGFBP-1 and -2, and reductions in free

  18. Colorimetric protein assay techniques.

    Science.gov (United States)

    Sapan, C V; Lundblad, R L; Price, N C

    1999-04-01

    There has been an increase in the number of colorimetric assay techniques for the determination of protein concentration over the past 20 years. This has resulted in a perceived increase in sensitivity and accuracy with the advent of new techniques. The present review considers these advances with emphasis on the potential use of such technologies in the assay of biopharmaceuticals. The techniques reviewed include Coomassie Blue G-250 dye binding (the Bradford assay), the Lowry assay, the bicinchoninic acid assay and the biuret assay. It is shown that each assay has advantages and disadvantages relative to sensitivity, ease of performance, acceptance in the literature, accuracy and reproducibility/coefficient of variation/laboratory-to-laboratory variation. A comparison of the use of several assays with the same sample population is presented. It is suggested that the most critical issue in the use of a chromogenic protein assay for the characterization of a biopharmaceutical is the selection of a standard for the calibration of the assay; it is crucial that the standard be representative of the sample. If it is not possible to match the standard with the sample from the perspective of protein composition, then it is preferable to use an assay that is not sensitive to the composition of the protein such as a micro-Kjeldahl technique, quantitative amino acid analysis or the biuret assay. In a complex mixture it might be inappropriate to focus on a general method of protein determination and much more informative to use specific methods relating to the protein(s) of particular interest, using either specific assays or antibody-based methods. The key point is that whatever method is adopted as the 'gold standard' for a given protein, this method needs to be used routinely for calibration.

  19. Comparison of real-time SYBR green dengue assay with real-time taqman RT-PCR dengue assay and the conventional nested PCR for diagnosis of primary and secondary dengue infection

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    Damodar Paudel

    2011-01-01

    Full Text Available Background : Dengue fever and dengue hemorrhagic fever are caused by dengue virus. Dengue infection remains a burning problem of many countries. To diagnose acute dengue in the early phase we improve the low cost, rapid SYBR green real time assay and compared the sensitivity and specificity with real time Taqman® assay and conventional nested PCR assay. Aims: To develop low cost, rapid and reliable real time SYBR green diagnostic dengue assay and compare with Taqman real-time assay and conventional nested PCR (modified Lanciotti. Materials and Methods: Eight cultured virus strains were diluted in tenth dilution down to undetectable level by the PCR to optimize the primer, temperature (annealing, and extension and to detect the limit of detection of the assay. Hundred and ninety three ELISA and PCR proved dengue clinical samples were tested with real time SYBR® Green assay, real time Taqman® assay to compare the sensitivity and specificity. Results: Sensitivity and specificity of real time SYBR® green dengue assay (84% and 66%, respectively was almost comparable to those (81% and 74% of Taqman real time PCR dengue assay. Real time SYBR® green RT-PCR was equally sensitive in primary and secondary infection while real time Taqman was less sensitive in the secondary infection. Sensitivity of real time Taqman on DENV3 (87% was equal to SYBR green real time PCR dengue assay. Conclusion: We developed low cost rapid diagnostic SYBR green dengue assay. Further study is needed to make duplex primer assay for the serotyping of dengue virus.

  20. Comparison of using polyurethane foam passive samplers and tree bark samples from Western China to determine atmospheric organochlorine pesticide.

    Science.gov (United States)

    Li, Qiuxu; Lu, Yao; Jin, Jun; Li, Guangyao; Li, Peng; He, Chang; Wang, Ying

    2016-03-01

    Polyurethane foam (PUF) passive samplers were deployed and tree bark samples were collected at 15 sites across western China in 2013, and the organochlorine pesticide (OCP) concentrations in the samples were determined. Dichlorodiphenyltrichloroethane and its degradation products (collectively called DDTs), hexachlorocyclohexanes (HCHs), and hexachlorobenzene (HCB) were the dominant OCPs in the PUF samples and tree bark samples. The mean DDTs, HCHs and HCB concentrations were 33, 22 and 18ng/sample in the PUF samples, and 428, 74, and 43ng/(g lipid weight (lw)) in the tree bark, respectively. The OCP concentrations in the air, calculated using PUF-air and tree-bark-air partitioning models, were of the same order of magnitude. Both sample types showed that relatively fresh inputs of DDT and HCHs to the environment have occurred in western China. Meanwhile, PUF passive samplers were compared with the use of tree bark samples as passive samplers. The OCP compositions in the PUF and tree bark samples were different. Only the relatively stable OCPs (such as HCB, β-HCH and p,p'-dichlorodiphenyldichloro-ethylene (DDE)) were consistent in the PUF and tree bark samples.

  1. Comparison of a flow assay for brucellosis antibodies with the reference cELISA test in West African Bos indicus.

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    Barend M deC Bronsvoort

    Full Text Available Brucellosis is considered by the Food and Agricultural Organisation and the World Health Organisation as one of the most widespread zoonoses in the world. It is a major veterinary public health challenge as animals are almost exclusively the source of infection for people. It is often undiagnosed in both human patients and the animal sources and it is widely acknowledged that the epidemiology of brucellosis in humans and animals is poorly understood, particularly in sub-Saharan Africa. It is therefore important to develop better diagnostic tools in order to improve our understanding of the epidemiology and also for use in the field for disease control and eradication. As with any new diagnostic test, it is essential that it is validated in as many populations as possible in order to characterise its performance and improve the interpretation of its results. This paper describes a comparison between a new lateral flow assasy (LFA for bovine brucellosis and the widely used cELISA in a no gold standard analysis to estimate test performance in this West African cattle population. A Bayesian formulation of the Hui-Walter latent class model incorporated previous studies' data on sensitivity and specificity of the cELISA. The results indicate that the new LFA is very sensitive (approximately 87% and highly specific (approximately 97%. The analysis also suggests that the current cut-off of the cELSIA may not be optimal for this cattle population but alternative cut-offs did not significantly change the estimates of the LFA. This study demonstrates the potential usefulness of this simple to use test in field based surveillance and control which could be easily adopted for use in developing countries with only basic laboratory facilities.

  2. Secular variation in Western Europe during the first millennium BC New full vector data and comparison with geomagnetic models

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    Hervé, G.; Chauvin, A.; Lanos, P.

    2011-12-01

    Archaeological structures in Western Europe are the most-used material to estimate the secular variation of the geomagnetic field during the last millennia. However there is still a lack of data especially for archaeointensities beyond the transition BC/AD, whereas already published data suggest very strong secular variation during the first millennium BC. This study presents 37 new archaeodirections and 18 new archaeointensities from France for the last 1500 years BC. Studied materials are kilns, hearths and two sets of pottery collections. Usual rock magnetism methods have been carried out to characterize magnetic grains. Archaeodirections were obtained by thermal and alternating fields demagnetization and they were corrected for thermal remanent magnetization anisotropy effects. Archaeointensities were determined with the classical Thellier-Thellier protocol with pTRM checks and take account of anisotropy and cooling rate effects. New Bayesian Western Europe secular variation curves for archaeodirection and archaeointensity were built with this new dataset and previously published data selected following reliability criteria. New curves present small variations of inclination during the last 1500 years BC. However for declination a very sharp maximum is observed around 800-750BC. Our new high-quality data set reveals also a regular decrease of archaeointensity between 800BC and the end of the first millennium BC. Our secular variation curves for France are very coherent with predicted directions computed with ARCH3K_cst.1 constrained model (Korte et al., 2009), but we note some discrepancies for archaeointensity between data and predicted values. ARCH3K_cst.1 constrained model built with archaeomagnetic and volcanic data seems more efficient than CALS3k.4 model (Korte & Constable, 2011), which includes archaeomagnetic, volcanic and sedimentary records. This study demonstrates consequently the central part of high-quality archaeomagnetic and volcanic data in the

  3. Examining cultural drifts in artworks through history and development: cultural comparisons between Japanese and western landscape paintings and drawings.

    Science.gov (United States)

    Nand, Kristina; Masuda, Takahiko; Senzaki, Sawa; Ishii, Keiko

    2014-01-01

    Research on cultural products suggest that there are substantial cultural variations between East Asian and European landscape masterpieces and contemporary members' landscape artwork (Masuda et al., 2008c), and that these cultural differences in drawing styles emerge around the age of 8 (Senzaki et al., 2014b). However, culture is not static. To explore the dynamics of historical and ontogenetic influence on artistic expressions, we examined (1) 17-20th century Japanese and Western landscape masterpieces, and (2) cross-sectional adolescent data in landscape artworks alongside previous findings of elementary school-aged children, and undergraduates. The results showed cultural variations in artworks and masterpieces as well as substantial "cultural drifts" (Herskovits, 1948) where at certain time periods in history and in development, people's expressions deviated from culturally default patterns but occasionally returned to its previous state. The bidirectional influence of culture and implications for furthering the discipline of cultural psychology will be discussed.

  4. Examining cultural drifts in artworks through history and development: cultural comparisons between Japanese and western landscape paintings and drawings

    Science.gov (United States)

    Nand, Kristina; Masuda, Takahiko; Senzaki, Sawa; Ishii, Keiko

    2014-01-01

    Research on cultural products suggest that there are substantial cultural variations between East Asian and European landscape masterpieces and contemporary members' landscape artwork (Masuda et al., 2008c), and that these cultural differences in drawing styles emerge around the age of 8 (Senzaki et al., 2014b). However, culture is not static. To explore the dynamics of historical and ontogenetic influence on artistic expressions, we examined (1) 17–20th century Japanese and Western landscape masterpieces, and (2) cross-sectional adolescent data in landscape artworks alongside previous findings of elementary school-aged children, and undergraduates. The results showed cultural variations in artworks and masterpieces as well as substantial “cultural drifts” (Herskovits, 1948) where at certain time periods in history and in development, people's expressions deviated from culturally default patterns but occasionally returned to its previous state. The bidirectional influence of culture and implications for furthering the discipline of cultural psychology will be discussed. PMID:25285085

  5. Examining Cultural Drifts in Artworks through History and Development: Cultural Comparisons between Japanese and Western Landscape Paintings and Drawings.

    Directory of Open Access Journals (Sweden)

    Kristina eNand

    2014-09-01

    Full Text Available Research on cultural products suggest that there are substantial cultural variations between East Asian and European landscape masterpieces and contemporary members’ landscape artwork (Masuda et al., 2008, and that these cultural differences in drawing styles emerge around the age of 8 (Senzaki et al., 2014. However, culture is not static. To explore the dynamics of historical and ontogenetic influence on artistic expressions, we examined (1 17th to 20th century Japanese and Western landscape masterpieces, and (2 cross-sectional adolescent data in landscape artworks alongside previous findings of elementary school-aged children, and undergraduates. The results showed cultural variations in artworks and masterpieces as well as substantial cultural drifts (Herskovits, 1948 where at certain time periods in history and in development, people’s expressions deviated from culturally default patterns but occasionally returned to its previous state. The bidirectional influence of culture and implications for furthering the discipline of cultural psychology will be discussed.

  6. The dynamics and energetics of midlatitude western boundary currents: A comparison of the Kuroshio Extension and the Gulf Stream

    Science.gov (United States)

    Mitchell, James L.; Hallock, Z. R.; Hurlburt, H. E.; Johnson, D. R.; Kindle, J. C.; Teague, W. J.; Thompson, J. D.; Schmitz, W. J.

    1991-01-01

    We will use TOPEX/POSEIDON altimetry in combination with ongoing and planned efforts, including data from the European Remote Sensing satellite (ERS-1), in situ measurements designed specifically to complement satellite altimetry, and assimilation of these several data types into eddy-resolving numerical models in order to understand the dynamics and energetics of midlatitude western boundary currents (specifically, the Gulf Stream and the Kuroshio Extension). A better understanding of the recirculation of midlatitude gyres can best be undertaken in the format of such regional, eddy-resolving experiments. Such regional programs will enhance and be enhanced by the larger scale circulation studies of the World Ocean Circulation Experiment (WOCE) as well as by global-scale eddy-resolving models that we will develop prior to the TOPEX/POSEIDON mission. This effort includes participation on the TOPEDX/POSEIDON Science Team.

  7. Comparison of laryngeal mask airway use with endotracheal intubation during anesthesia of western lowland gorillas (Gorilla gorilla gorilla).

    Science.gov (United States)

    Cerveny, Shannon N; D'Agostino, Jennifer J; Davis, Michelle R; Payton, Mark E

    2012-12-01

    The laryngeal mask airway is an alternative to endotracheal intubation that achieves control of the airway by creating a seal around the larynx with an inflatable cuff. This study compared use of the laryngeal mask airway with endotracheal intubation in anesthetized western lowland gorillas (Gorilla gorilla gorilla). Eight adult gorillas were immobilized for routine and diagnostic purposes for a total of nine anesthetic events. During each anesthetic event, gorillas were either intubated (n = 4; group A) or fitted with a laryngeal mask airway (n= 5; group B). Time required to place each airway device, physiologic parameters, and arterial blood gas were measured and compared between the two groups. There were no significant differences between the two groups for time required to place airway device, heart rate, hemoglobin oxygen saturation, end-tidal carbon dioxide, arterial partial pressure of carbon dioxide, or arterial pH between the two groups. Mean arterial partial pressure of oxygen was significantly greater in group B, 15 (group A: 94 +/- 44 mm Hg; group B: 408 +/- 36 mm Hg; P= 0.0025) and 45 (group A: 104 +/- 21 mm Hg; group B: 407 +/- 77 mm Hg; P = 0.0026) min after airway device placement. Mean respiratory rate was significantly greater in group A at multiple time points. Mean arterial pressure (group A: 129 +/- 16 mm Hg; group B: 60 +/- 8 mm Hg) and diastolic blood pressure (group A: 115 +/- 21 mm Hg; group B: 36 +/- 10 mm Hg) were significantly greater in group A at the time of airway device placement. The laryngeal mask airway maintained oxygenation and ventilation effectively in all gorillas and is a useful alternative to endotracheal intubation in western lowland gorillas.

  8. Comparison of a commercial real-time PCR assay, RealCycler® PJIR kit, progenie molecular, to an in-house real-time PCR assay for the diagnosis of Pneumocystis jirovecii infections.

    Science.gov (United States)

    Guillaud-Saumur, Thibaud; Nevez, Gilles; Bazire, Amélie; Virmaux, Michèle; Papon, Nicolas; Le Gal, Solène

    2017-04-01

    We compared the RealCycler® PJIR kit (Progenie Molecular), available in Europe, to an in-house real-time PCR assay for the diagnosis of Pneumocystis jirovecii infections. Excellent agreement was found (concordance rate, 97.4%; Cohen's kappa, 0.918>0.8) showing that this commercial assay represents an alternative method for the diagnosis of P. jirovecii infections.

  9. Validation of a fluorescence polarization assay (FPA) performed in microplates and comparison with other tests used for diagnosing B. melitensis infection in sheep and goats.

    Science.gov (United States)

    Minas, A; Stournara, A; Minas, M; Stack, J; Petridou, E; Christodoulopoulos, G; Krikelis, V

    2007-03-30

    Fluorescence polarization assay (FPA) is a relatively new test for the serological diagnosis of Brucella spp. infection in animals. FPA, carried out in 96-well microplate format, was validated here for diagnosing B. melitensis infection in sheep and goats. This study included sera from 1933 sheep and goats, from animals reared in naturally infected flocks (verified by culture) and showing a positive reaction to two different tests conducted in parallel. In addition, 2154 sera originating from healthy sheep and goats, reared in areas where B. melitensis had never been isolated, were assayed. The optimum cut-off value offering the highest diagnostic sensitivity (DSn) and diagnostic specificity (DSp) was determined at 15 mP over the mean value of the buffer control used in each microplate as determined by receiver operating characteristic analysis. The DSn and DSp of the FPA for small ruminants carried out in microplates at this cut-off value were calculated to be 95.9% and 97.9% with 95% confidence intervals (95% CI) of 94.9-97.7% and 97.2-98.4%, respectively. The accuracy of the FPA, as expressed by determination of the area under the curve, was 0.991. Indirect ELISA and FPA tests offered the highest DSn when compared with the Rose Bengal test, the complement fixation test, the modified Rose Bengal test and competitive ELISA. The parallel or serial combination of FPA with indirect ELISA offers the highest DSn and DSp. As temperature can affect the results of the FPA, all reagents must be at the same temperature and the standard for comparison must always be read under the same conditions as the sera under test. FPA performed in microplates is a promising assay; the DSn and accuracy are better than those of the tests currently approved for diagnosing B. melitensis in small ruminants. Because of its simplicity, speed, and accuracy, this test can improve capacity for laboratory testing and the efficacy of an eradication program based on a test-and-slaughter policy.

  10. UPLC-MRM Mass Spectrometry Method for Measurement of the Coagulation Inhibitors Dabigatran and Rivaroxaban in Human Plasma and Its Comparison with Functional Assays.

    Directory of Open Access Journals (Sweden)

    Joachim Kuhn

    least one week. A method comparison between our UPLC-MRM MS method, the commercially available automated Direct Thrombin Inhibitor assay (DTI assay for dabigatran measurement from CoaChrom Diagnostica, as well as the automated anti-Xa assay for rivaroxaban measurement from Chromogenix both performed by ACL-TOP showed a high degree of correlation. However, UPLC-MRM MS measurement of dabigatran and rivaroxaban has a much better selectivity than classical functional assays measuring activities of various coagulation factors which are susceptible to interference by other coagulant drugs.Overall, we developed and validated a sensitive and specific UPLC-MRM MS assay for the quick and specific measurement of dabigatran and rivaroxaban in human plasma.

  11. Comparison of major antigenic proteins of six strains of the human granulocytic ehrlichiosis agent by Western immunoblot analysis.

    Science.gov (United States)

    Zhi, N; Rikihisa, Y; Kim, H Y; Wormser, G P; Horowitz, H W

    1997-10-01

    The etiologic agent of human granulocytic ehrlichiosis (HGE) is an obligate intracellular bacterium. In 1996, blood specimens from 53 patients suspected of having HGE were examined by indirect fluorescent antibody (IFA) testing with the HGE agent no. 13 isolate as the antigen, by nested PCR, and by culture. All patients resided in Westchester County, N.Y. Twelve patient specimens were positive for IFA (titer > or = 1:40). Seven of these were also positive by PCR. Of the seven specimens positive by both IFA testing and PCR, the HGE agent was isolated from four (no. 2, 3, 6, and 11) and continuously cultured in HL-60 cells. These were confirmed as the HGE agent by sequencing of 16S rDNA. Both purified whole-cell organisms and the outer membrane fractions of the new isolates were compared with no. 13 isolate and a tick (USG) isolate by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western immunoblot analysis. No. 11 and 13 isolates had identical SDS-PAGE patterns with respect to 49- and 47-kDa proteins. No. 3 and USG isolates lacked the 47-kDa protein, and no. 6 isolate lacked the 49-kDa protein. Both 49- and 47-kDa bands were absent in no. 2 isolate. Western blot results with seven different sera, including five convalescent-phase sera from these patients, one dog anti-USG isolate, and one horse anti-BDS isolate, showed that all major antigens in six isolates were recognized by all sera. However, the molecular sizes and the numbers of major antigens recognized varied among the six isolates. Overall, HGE agent no. 3, 6, 11, and 13, and USG isolates had similar patterns, with 1 or 2 major antigens with molecular masses of around 49 and 47 kDa. No. 2 isolate was quite distinct in having a major antigen of 43 kDa. This indicates that although these antigenic epitopes are all cross-reactive among strains, the HGE agent has a strain pleomorphism in its major antigenic proteins. The major antigen profiles of the outer membrane protein fractions

  12. Western psychology and Muslim psychology in dialogue: comparisons between a Qura'nic theory of personality and Freud's and Jung's ideas.

    Science.gov (United States)

    Abu-Raiya, Hisham

    2014-04-01

    In this paper, comparisons are made between a newly developed Qura'nic theory of personality and the Freudian and Jungian theories of the mind. Notable similarities were found between the Freudian id, ego, superego and neurosis and the Qura'nic nafs ammarah besoa' (evil-commanding psyche), a'ql (intellect), al-nafs al-lawammah (the reproachful psyche) and al-nafs al-marid'a (the sick psyche), respectively. Noteworthy resemblances were detected also between the Jungian concepts collective unconscious, archetypes, Self and individuation and the Qura'nic constructs roh (spirit), al-asmaa' (the names), qalb (heart), and al-nafs al-mutmainnah (the serene psyche), respectively. These parallels, as well as the departure points, between the models are thoroughly discussed and analyzed. The comparisons performed in this paper open new avenues for dialogue between western models of the psyche and their Muslim counterparts, a dialogue that can enrich both perspectives and advance the field of psychology.

  13. Comparison of GP5+/6+-PCR and SPF10-line blot assays for detection of high-risk human papillomavirus in samples from women with normal cytology results who develop grade 3 cervical intraepithelial neoplasia.

    NARCIS (Netherlands)

    Hesselink, A.T.; Ham, M.A.P.C. van; Heideman, D.A.; Groothuismink, Z.M.; Rozendaal, L.; Berkhof, J.; Kemenade, F.J. van; Massuger, L.A.; Melchers, W.J.G.; Meijer, C.J.; Snijders, P.J.L.M.

    2008-01-01

    Using a case control approach, we performed a two-way comparison study between GP5+/6+-PCR and HPV SPF(10)-Line Blot 25 (SPF(10)) assays for detection of 14 types of high-risk human papillomavirus (hrHPV) in samples from women with normal cytology results who had or developed grade 3 cervical

  14. Comparison of Ocean Dynamics with a Regional Circulation Model and Improved Altimetry in the North-Western Mediterranean

    Directory of Open Access Journals (Sweden)

    Jérôme Bouffard

    2008-01-01

    Full Text Available The spatial and temporal resolution of satellite altimetry is usually sufficient for monitoring the changes of sea surface topography in the open ocean. However, coastal ocean dynamics are much more complex, being characterized by smaller spatial and temporal scales of variability. The quality and availability of satellite-derived products along the coasts have to be improved, with a strategy optimized for coastal targets. Therefore a coastal multi-satellite altimetry dataset (TOPEX/Poseidon, Jason-1; Envisat; GFO at a 10 - 20 Hz sampling rate has been derived from routine geophysical data products using a new processing software dedicated to coastal zone applications. Improved along-track sea level variations with fine space scales are available in the North-western Mediterranean Sea from 2001 to 2003, and are compared with high-resolution numerical model elevations from the eddy-resolving model SYMPHONIE. This preparatory work emphasizes the potential of improved multi-satellite altimetry for validating coastal hydro-dynamical models and could contribute in the future to a better tuning of the boundary conditions of the simulations.

  15. Comparison of Indo-Pacific humpback dolphin (Sousa chinensis) whistles from two areas of western Peninsular Malaysia.

    Science.gov (United States)

    Hoffman, Jordan M; Ponnampalam, Louisa S; Araújo, Claryana C; Wang, John Y; Kuit, Sui Hyang; Hung, Samuel K

    2015-11-01

    Differences in the acoustic variables of whistles emitted by Indo-Pacific humpback dolphins (Sousa chinensis) from two coastal locations along western Peninsular Malaysia were investigated. Duration, frequency, and frequency modulation variables were extracted from and used to characterize recordings of free-ranging humpback dolphins that were made using a broadband towed hydrophone. A total of 960 whistles from Matang Mangroves and 823 whistles from Langkawi Island were used in analyses. The whistles of Malaysian humpback dolphins covered frequencies from 1231 to 27 120 Hz with durations from 0.010-1.575 s. Significant multivariate differences were found in whistles emitted between locations. Significant differences were also found between dolphins of the two locations in their whistle duration, frequency modulation, and all frequency variables except for minimum frequency, which is likely under morphological constraints. The differences in whistles may be related to adaptations to the local acoustic habitat or unique whistles may have developed due to social interactions within each location, or broader scale differences resulting from geographic separation between the locations.

  16. Platform margins, reef facies, and microbial carbonates; a comparison of Devonian reef complexes in the Canning Basin, Western Australia, and the Guilin region, South China

    Science.gov (United States)

    Shen, Jian-Wei; Webb, Gregory E.; Jell, John S.

    2008-05-01

    Devonian reef complexes were well developed in Western Australia and South China, but no detailed direct comparison has been made between reef building in the two regions. The regions differ in several respects, including tectonic, stratigraphic and palaeoceanographic-palaeogeographic settings, and the reef building styles reflect minor differences in reef builders and reef facies. Similarities and differences between the two reef complexes provide insights into the characteristics of platform margins, reef facies and microbial carbonates of both regions. Here we present a comparison of platform margin types from different stratigraphic positions in the Late Devonian reef complex of the Canning Basin, Western Australia and Middle and Late Devonian margin to marginal slope successions in Guilin, South China. Comparisons are integrated into a review of the reefal stratigraphy of both regions. Reef facies, reef complex architecture, temporal reef builder associations, 2nd order stratigraphy and platform cyclicity in the two regions were generally similar where the successions overlap temporally. However, carbonate deposition began earlier in South China. Carbonate complexes were also more widespread in South China and represent a thicker succession overall. Platforms in the Canning Basin grew directly on Precambrian crystalline basement or early Palaeozoic sedimentary rocks, but in South China, carbonate complexes developed conformably on older Devonian siliciclastic strata. Pre-Frasnian reef facies in South China had more abundant skeletal frameworks than in Canning Basin reefs of equivalent age, and Famennian shoaling margins containing various microbial reefs may have been more common and probably more diverse in South China. However, Late Devonian platform margin types have been documented more completely in the Canning Basin. Deep intra-platform troughs (deep depressions containing non-carbonate pelagic sediments — Nandan-type successions) that developed along

  17. Tectonically driven fluid flow and gold mineralisation in active collisional orogenic belts: comparison between New Zealand and western Himalaya

    Science.gov (United States)

    Craw, D.; Koons, P. O.; Horton, T.; Chamberlain, C. P.

    2002-04-01

    Hydrothermal activity and mesothermal-styled gold mineralisation occurs near the main topographic divide of most active or young collisional mountain belts. The Southern Alps of New Zealand is used in this study as a model for the mineralising processes. The collisional tectonics results in a two-sided wedge-shaped orogen into which rock is transported horizontally. Upper crustal rocks pass through the orogen and leave the orogen by erosion, whereas lower crustal rocks are deformed into the mountain roots. High relief drives meteoric water flow to near the brittle-ductile transition. Lower to upper greenschist facies metamorphic reactions, driven by deformation at the crustal decollement and in the root, release water-rich fluids that rise through the orogen. Intimate chemical interaction between fluid and rock results in dissolution and later precipitation of gold, arsenic and sulphur. Fluid flow and mineralisation in the topographic divide region is facilitated by a network of steeply dipping faults and associated rock damage zones where oblique strike-slip faults intersect the thrust faults that strike subparallel to the main mountain range. The Nanga Parbat massif of the western Himalaya is an example of an active collisional zone which hosts hydrothermal activity but no gold mineralisation. The lack of gold mineralisation is due to the following factors: CO 2-dominated rising metamorphic fluid in dehydrated amphibolite-granulite facies metamorphic rocks does not dissolve gold and arsenic; hot (up to 400 °C) meteoric water confined to fractures in the gneiss limits dissolution of gold and arsenic; low density of hot water/dry steam, and low reduced sulphur content of fluid, restrict solubility of gold and arsenic; absence of fracture networks in the core of the massif and the small volumes of circulating fluid limit metal concentration; and lack of reactive rock compositions limits chemically mediated metal deposition.

  18. Comparison of soil bacterial communities of Pinus patula of Nilgiris, western ghats with other biogeographically distant pine forest clone libraries.

    Science.gov (United States)

    Rohini-Kumar, M; Osborne, Jabez W; Saravanan, V S

    2013-07-01

    The bacterial community structure of the rhizosphere and non-rhizosphere soil of Pinus patula, found in the Nilgiris region of Western Ghats, was studied by constructing 16S rRNA gene clone libraries. In the rhizosphere and non-rhizosphere soil clone libraries constructed, 13 and 15 bacterial phyla were identified, respectively. The clone libraries showed the predominance of members of culturally underrepresented phyla like Acidobacteria and Verrucomicrobia. The Alphaproteobacteria and Acidobacteria clones were predominant in rhizosphere and non-rhizosphere soil samples, respectively. In rhizosphere, amongst Alphaproteobacteria members, Bradyrhizobium formed the significant proportion, whereas in non-rhizosphere, members of subdivision-6 of phylum Acidobacteria were abundant. The diversity analysis of P. patula soil libraries showed that the phylotypes (16S rRNA gene similarity cutoff, ≥97 %) of Acidobacteria and Bacteroidetes were relatively predominant and diverse followed by Alphaproteobacteria and Verrucomicrobia. The diversity indices estimated higher richness and abundance of bacteria in P. patula soil clone libraries than the pine forest clone libraries retrieved from previous studies. The tools like principal co-ordinate analysis and Jackknife cluster analysis, which were under UniFrac analysis indicated that variations in soil bacterial communities were attributed to their respective geographical locations due to the phylogenetic divergence amongst the clone libraries. Overall, the P. patula rhizosphere and non-rhizosphere clone libraries were found significantly unique in composition, evenly distributed and highly rich in phylotypes, amongst the biogeographically distant clone libraries. It was finally hypothesised that the phylogenetic divergence amongst the bacterial phylotypes and natural selection plays a pivotal role in the variations of bacterial communities across the geographical distance.

  19. The relationship between mammal faunas and climatic instability since the Last Glacial Maximum: A Nearctic vs. Western Palearctic comparison

    Science.gov (United States)

    Torres-Romero, Erik Joaquín; Varela, Sara; Fisher, Jason T.; Olalla-Tárraga, Miguel Á.

    2017-07-01

    Climate has played a key role in shaping the geographic patterns of biodiversity. The imprint of Quaternary climatic fluctuations is particularly evident on the geographic distribution of Holarctic faunas, which dramatically shifted their ranges following the alternation of glacial-interglacial cycles during the Pleistocene. Here, we evaluate the existence of differences between climatically stable and unstable regions - defined on the basis of climatic change velocity since the Last Glacial Maximum - in the geographic distribution of several biological attributes of extant terrestrial mammals of the Nearctic and Western Palearctic regions. Specifically, we use a macroecological approach to assess the dissimilarities in species richness, range size, body size, longevity and litter size of species that inhabit regions with contrasting histories of climatic stability. While several studies have documented how the distributional ranges of animals can be affected by long-term historic climatic fluctuations, there is less evidence on the species-specific traits that determine their responsiveness under such climatic instability. We find that climatically unstable areas have more widespread species and lower mammal richness than stable regions in both continents. We detected stronger signatures of historical climatic instability on the geographic distribution of body size in the Nearctic region, possibly reflecting lagged responses to recolonize deglaciated regions. However, the way that animals respond to climatic fluctuations varies widely among species and we were unable to find a relationship between climatic instability and other mammal life-history traits (longevity and litter size) in any of the two biogeographic regions. We, therefore, conclude that beyond some biological traits typical of macroecological analyses such as geographic range size and body size, it is difficult to infer the responsiveness of species distributions to climate change solely based on

  20. Comparison of a commercial ELISA and an immunoperoxidase monolayer assay to detect antibodies directed against porcine respiratory and reproductive syndrome virus

    NARCIS (Netherlands)

    Nodelijk, G.; Wensvoort, G.; Kroese, B.; Leengoed, van L.A.M.G.; Colijn, E.; Verheijden, J.H.M.

    1996-01-01

    A commercially available enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies against porcine respiratory and reproductive syndrome virus (PRRSV) was compared to an immunoperoxidase monolayer assay (IPMA). Serum samples used were collected from pigs experimentally infected with

  1. Comparison of gene expression regulation in mouse- and human embryonic stem cell assays during neural differentiation and in response to valproic acid exposure

    NARCIS (Netherlands)

    Schulpen, Sjors H. W.; Theunissen, Peter T.; Pennings, Jeroen L. A.; Piersma, Aldert H.

    2015-01-01

    Embryonic stem cell tests (EST) are considered promising alternative assays for developmental toxicity testing. Classical mouse derived assays (mEST) are being replaced by human derived assays (hEST), in view of their relevance for human hazard assessment. We have compared mouse and human neural EST

  2. Auto-antibodies to double-stranded DNA as biomarker in systemic lupus erythematosus : comparison of different assays during quiescent and active disease

    NARCIS (Netherlands)

    de Leeuw, Karina; Bungener, Laura; Roozendaal, Caroline; Bootsma, Hendrika; Stegeman, Coen A

    2017-01-01

    OBJECTIVE: Auto-antibodies directed to dsDNA (anti-dsDNA) are used in diagnosis and follow-up for SLE. However, multiple assays are used. The objective of this study was to determine the best-performing assays, especially in prediction of exacerbations. METHODS: Seven assays were compared during LN

  3. Comparison of Roche MONITOR and Organon Teknika NucliSens assays to quantify human immunodeficiency virus type 1 RNA in cerebrospinal fluid.

    Science.gov (United States)

    Spearman, P; Fiscus, S A; Smith, R M; Shepard, R; Johnson, B; Nicotera, J; Harris, V L; Clough, L A; McKinsey, J; Haas, D W

    2001-04-01

    We compared Roche MONITOR and Organon Teknika NucliSens assays for human immunodeficiency virus type 1 (HIV-1) RNA in cerebrospinal fluid (CSF). Results of 282 assays were highly correlated (r = 0.826), with MONITOR values being 0.29 +/- 0.4 log(10) copies/ml (mean +/- standard deviation) values. Both assays can reliably quantify HIV-1 RNA in CSF.

  4. Comparison of Gull Feces-specific Assays Targeting the 16S rRNA Gene of Catellicoccus Marimammalium and Streptococcus spp.

    Science.gov (United States)

    Two novel gull-specific qPCR assays were developed using 16S rRNA gene sequences from gull fecal clone libraries: a SYBR-green-based assay targeting Streptococcus spp. (i.e., gull3) and a TaqMan qPCR assay targeting Catellicoccus marimammalium (i.e., gull4). The main objectives ...

  5. Comparison of the Luminex xTAG RVP Fast assay and the Idaho Technology FilmArray RP assay for detection of respiratory viruses in pediatric patients at a cancer hospital.

    Science.gov (United States)

    Babady, N Esther; Mead, Peter; Stiles, Jeffrey; Brennan, Carrie; Li, Haijing; Shuptar, Susan; Stratton, Charles W; Tang, Yi-Wei; Kamboj, Mini

    2012-07-01

    Respiratory viruses are increasingly recognized as serious causes of morbidity and mortality in immunocompromised patients. The rapid and sensitive detection of respiratory viruses is essential for the early diagnosis and administration of appropriate antiviral therapy, as well as for the effective implementation of infection control measures. We compared the performance of two commercial assays, xTAG RVP Fast (Luminex Diagnostics, Toronto, Canada) and FilmArray RVP (FA RVP; Idaho Technology, Salt Lake City, UT), in pediatric patients at Memorial Sloan-Kettering Cancer Center. These assays detect the following viruses: respiratory syncytial virus; influenza A and B viruses; parainfluenza viruses 1, 2, 3, and 4; human metapneumovirus; adenovirus; enterovirus-rhinovirus; coronaviruses NL63, HKU1, 229E, and OC43; and bocavirus. We tested a total of 358 respiratory specimens from 173 pediatric patients previously tested by direct fluorescence assay (DFA) and viral culture. The overall detection rate (number of positive specimens/total specimens) for viruses tested by all methods was 24% for DFA/culture, 45% for xTAG RVP Fast, and 51% for FA RVP. The agreement between the two multiplex assays was 84.5%, and the difference in detection rate was statistically significant (P < 0.0001). Overall, the FA RVP assay was more sensitive than the xTAG RVP Fast assay and had a turnaround time of approximately 1 h. The sensitivity, simplicity, and random-access platform make FA RVP an excellent choice for laboratory on-demand service with low to medium volume.

  6. 新孢子虫和弓形虫ELISA及western blot检测方法的建立及应用%Establishment and application of indirect ELISA assays and western blot to detect antibodies agaist Neospora caninum and Toxoplasma gondii

    Institute of Scientific and Technical Information of China (English)

    陈亮; 闫双; 刘启生; 曹雯丽; 王真; 巴音查汗

    2012-01-01

    为建立牛新孢子虫和弓形虫的免疫学检测方法,并调查新疆部分地区牛新孢子虫和弓形虫的感染情况,本研究应用纯化的新孢子虫重组蛋白SRS2 (NcSRS2)和弓形虫重组蛋白SAG2 (TgSAG2)作为包被抗原,分别建立新孢子虫和弓形虫的ELISA和western blot血清学检测方法,并进行特异性和重复性试验,以其检测662份疑似样品,并与商品化试剂盒检测结果比较验证.特异性和重复性试验结果表明,建立的方法特异性强、重复性良好.采用建立的两种ELISA方法对662份临床样品的检测结果表明,新孢子虫和弓形虫的抗体阳性率分别为13.44%(89/662)和5.29%(35/662);与IDEXX试剂盒和永辉试剂盒的符合率分别为94.11%和95.92%.此外,western blot检测的新孢子虫和弓形虫抗体阳性率分别为5.14%(34/662)和3.17%(21/662);与建立的ELISA检测方法的符合率分别为91.69%和97.89%.本研究为分析奶牛流产的原因提供了一定的依据.%Neospora caninum and Toxoplasma gondii are closely related protozoan parasites which causes of abortion and congenital disease in ruminants. To establish the methods for detection of the antibodies against N. caninum and T. gondii in bovine, The indirect ELISAs and western blot assay were developed based on purified recombinant protein SRS2 of N. cammim and SAG2 of T. gondii, respectively. The results show that these methods were highly specificity and repeatability. A total of 662 bovine serum samples were tested and the positive rates for N. caninum and T. gondii were 13.44% (89/662) and 5.29% (35/662) detected by the developed ELISAs, which were 94.11% and 95.92% agreements with TDEXX' N. caninum Antibody Test Kit and Yong hui' T. gondii Antibody Test Kit, respectively. In addition, the positive rates for N. caninum and T. gondii were 5.14% (34/662) and 3.17% (21/662) using western blot, and the agreements with the developed ELISAs were 91.69% and 97

  7. Comparison of Mercury Mass Loading in Streams to Wet and Dry Atmospheric Deposition in Watersheds of the Western US: Evidence for Non-Atmospheric Mercury Sources

    Science.gov (United States)

    Domagalski, J. L.; Majewski, M. S.; Alpers, C. N.; Eckley, C.

    2015-12-01

    Many streams in the western United States (US) are listed as impaired by mercury (Hg), and it is important to understand the magnitudes of the various sources in order to implement management strategies. Atmospheric deposition of Hg and can be a major source of aquatic contamination, along with mine wastes, and other sources. Prior studies in the eastern US have shown that streams deliver less than 50% of the atmospherically deposited Hg on an annual basis. In this study, we compared annual stream Hg loads for 20 watersheds in the western US to measured wet and modeled dry deposition. Land use varies from undisturbed to mixed (agricultural, urban, forested, mining). Data from the Mercury Deposition Network was used to estimate Hg input from precipitation. Dry deposition was not directly measured, but can be modeled using the Community Multi-scale Air Quality model. At an undeveloped watershed in the Rocky Mountains, the ratio of stream Hg load to atmospheric deposition was 0.2 during a year of average precipitation. In contrast, at the Carson River in Nevada, with known Hg contamination from historical silver mining with Hg amalgamation, stream export exceeded atmospheric deposition by a factor of 60, and at a small Sierran watershed with gold mining, the ratio was 70. Larger watersheds with mixed land uses, tend to have lower ratios of stream export relative to atmospheric deposition suggesting storage of Hg. The Sacramento River was the largest watershed for which Hg riverine loads were available with an average ratio of stream Hg export to atmospheric deposition of 0.10. Although Hg was used in upstream historical mining operations, the downstream river Hg load is partially mitigated by reservoirs, which trap sediment. This study represents the first compilation of riverine Hg loads in comparison to atmospheric deposition on a regional scale; the approach may be useful in assessing the relative importance of atmospheric and non-atmospheric Hg sources.

  8. What is the P value of Siberian soils? Soil phosphorus status in south-western Siberia and comparison with a global data set

    Science.gov (United States)

    Brédoire, Félix; Bakker, Mark R.; Augusto, Laurent; Barsukov, Pavel A.; Derrien, Delphine; Nikitich, Polina; Rusalimova, Olga; Zeller, Bernd; Achat, David L.

    2016-04-01

    Climate change is particularly strong in northern Eurasia and substantial ecological changes are expected in this extensive region. The reshaping and migration northwards of bioclimatic zones may offer opportunities for agricultural development in western and central Siberia. However, the bioclimatic vegetation models currently employed for projections still do not consider soil fertility, in spite of this being highly critical for plant growth. In the present study, we surveyed the phosphorus (P) status in the south-west of Siberia where soils have developed on loess parent material. We selected six sites differing in pedoclimatic conditions and the soil was sampled at different depths down to 1 m in aspen (Populus tremula L.) forest as well as in grassland areas. The P status was assessed by conventional methods and by isotope dilution kinetics. We found that P concentrations and stocks, as well as their distribution through the soil profile, were fairly homogeneous on the regional scale studied, although there were some differences between sites (particularly in organic P). The young age of the soils, together with slow kinetics of soil formation processes have probably not yet resulted in a sufficiently wide range of soil physico-chemical conditions to observe a more diverging P status. The comparison of our data set with similar vegetation contexts on the global scale revealed that the soils of south-western Siberia, and more generally of northern Eurasia, often have (very) high levels of total, organic and inorganic P. The amount of plant-available P in topsoils, estimated by the isotopically exchangeable phosphate ions, was not particularly high but was intermediate on the global scale. However, large stocks of plant-available P are stored in subsurface layers which currently have low fine-root exploration intensities. These results suggest that the P resource is unlikely to constrain vegetation growth and agricultural development under the present

  9. Risk factors for inadequate TB case finding in Rural Western Kenya: a comparison of actively and passively identified TB patients.

    Directory of Open Access Journals (Sweden)

    Anna H Van't Hoog

    Full Text Available BACKGROUND: The findings of a prevalence survey conducted in western Kenya, in a population with 14.9% HIV prevalence suggested inadequate case finding. We found a high burden of infectious and largely undiagnosed pulmonary tuberculosis (PTB, that a quarter of the prevalent cases had not yet sought care, and a low case detection rate. OBJECTIVE AND METHODS: We aimed to identify factors associated with inadequate case finding among adults with PTB in this population by comparing characteristics of 194 PTB patients diagnosed in a health facility after self-report, i.e., through passive case detection, with 88 patients identified through active case detection during the prevalence survey. We examined associations between method of case detection and patient characteristics, including HIV-status, socio-demographic variables and disease severity in univariable and multivariable logistic regression analyses. FINDINGS: HIV-infection was associated with faster passive case detection in univariable analysis (crude OR 3.5, 95% confidence interval (CI 2.0-5.9, but in multivariable logistic regression this was largely explained by the presence of cough, illness and clinically diagnosed smear-negative TB (adjusted OR (aOR HIV 1.8, 95% CI 0.85-3.7. Among the HIV-uninfected passive case detection was less successful in older patients aOR 0.76, 95%CI 0.60-0.97 per 10 years increase, and women (aOR 0.27, 95%CI 0.10-0.73. Reported current or past alcohol use reduced passive case detection in both groups (0.42, 95% CI 0.23-0.79. Among smear-positive patients median durations of cough were 4.0 and 6.9 months in HIV-infected and uninfected patients, respectively. CONCLUSION: HIV-uninfected patients with infectious TB who were older, female, relatively less ill, or had a cough of a shorter duration were less likely found through passive case detection. In addition to intensified case finding in HIV-infected persons, increasing the suspicion of TB among HIV

  10. Comparison of drier- to wetter-interval estuarine roof facies in the Eastern and Western Interior coal basins, USA

    Science.gov (United States)

    Archer, A.W.; Feldman, H.R.; Kvale, E.P.; Lanier, Wendy E.

    1994-01-01

    Many of the Carboniferous coals in the eastern interior of the US are associated with siliciclastic roof facies that were deposited within a fluvio-estuarine transition. These facies include a variety of rhythmites, some of which exhibit tidal cycles. Drier-interval coals (Westphalian B-C, Stephanian) tend to be more laterally restricted and more commonly are associated with paleovalleys. Conversely, wetter-interval coals (Westphalian D) are very widespread and are not restricted to paleovalleys. Throughout the Lake Carboniferous, wet paleoclimates associated with these coals lead to valley incision during sea-level lowstand when large tropical rivers downcut older sediments deposited during previous sea-level highstands. During subsequent rise of sea level, these fluvial valleys were flooded and converted to estuaries where tidal ranges and sedimentation rates were significantly amplified. Based on modern analogs and interpretation of many examples of Carboniferous tidal rhythmites, the localized depositional rates in these settings are exceptionally high. The estuaries became sediment sinks, trapping sediment that is pumped in from both fluvial and marine sources. As a result, sedimentation readily keeps pace with rising baselevel. Extensive intertidal flats and shallow subtidal flats are created and prograde over the valley-confined mires. Thick tidal cycles and upright trees (some with attached foliage) record rapid burial of mires. This model is supported with examples of roof facies from the Westphalian B-C of the Eastern Interior Basin, and the Stephanian of the Western Interior Basin. In these areas facies within each cycle range from well-developed, extensive paleosols and coals, to widespread marine shales or limestones. Variations in both sea level and climate resulted in a complex history of valley fill during which coals could be developed at any time (except during widespread flooding). Minable, low-sulfur and low-ash coals occur, but the coals are

  11. Comparison of antibody assays for detection of autoantibodies to Ro 52, Ro 60 and La associated with primary Sjögren's syndrome.

    Science.gov (United States)

    Trier, Nicole Hartwig; Nielsen, Inger Ødum; Friis, Tina; Houen, Gunnar; Theander, Elke

    2016-06-01

    Anti-Ro(52/60) and anti-La constitute the hallmark autoantibodies in primary Sjögren's syndrome, being present in 40-70% of sera. Several anti-Ro/La assays exist, but antibody detection appears to be assay-specific, thus the aim of this study was to compare several anti-Ro/La assays. In total, 96 sera from individuals with primary Sjögren's syndrome and 114 healthy controls were tested for anti-Ro 52/60 and anti-La in 17 immunoassays. Especially the immunoassays used for detection of anti-Ro 52 differed in their sensitivity (48-79%), while only small differences in sensitivities were observed for the anti-Ro 60 (69-77%) anti-La (39-44%) assays. Concordances of 65%, 79% and 73% for the anti-Ro 52, anti-Ro 60 and anti-La assays were found, respectively. The majority of the assays yielded high specificities, primarily ranging from 97 to 100%, except from a single anti-Ro 60 assay, which yielded a specificity of 79%. Occasionally, reactivity levels were increased in a few assays, indicating that false-positive results can be obtained when applying assays of reduced specificity. In general, the commercial assays appeared to perform better than the in-house analyses. When correcting the in-house assays for background reactivity, sensitivities were reduced by approximately 7%, 17%, and 19% for anti-Ro 52, anti-Ro 60 and anti-La assays, respectively, illustrating the pitfalls when applying immunoassays for detection of autoantibodies, which in theory may apply to commercial assays as well. Finally, increased total sensitivities were obtained when combining assays. These studies contribute to clarify the clinical utility of immunoassays for detection of autoantibodies of Ro 52, Ro 60 and La and illustrate that the most efficient strategy to maximize antibody sensitivity is to combine several assays.

  12. Western Sufism

    DEFF Research Database (Denmark)

    Sedgwick, Mark

    Western Sufism is sometimes dismissed as a relatively recent "new age" phenomenon, but in this book, Mark Sedgwick argues that it actually has very deep roots, both in the Muslim world and in the West. In fact, although the first significant Western Sufi organization was not established until 1915......, the first Western discussion of Sufism was printed in 1480, and Western interest in some of the ideas that are central to Sufi thought goes back to the thirteenth century. Sedgwick starts with the earliest origins of Western Sufism in late antique Neoplatonism and early Arab philosophy, and traces later...

  13. High resolution climate and vegetation simulations of the Late Pliocene, a model-data comparison over western Europe and the Mediterranean region

    Directory of Open Access Journals (Sweden)

    A. Jost

    2009-10-01

    Full Text Available Here we perform a detailed comparison between climate model results and climate reconstructions in western Europe and the Mediterranean area for the mid-Piacenzian warm interval (ca 3 Myr ago of the Late Pliocene epoch. This region is particularly well suited for such a comparison as several quantitative climate estimates from local pollen records are available. They show evidence for temperatures significantly warmer than today over the whole area, mean annual precipitation higher in northwestern Europe and equivalent to modern values in its southwestern part. To improve our comparison, we have performed high resolution simulations of the mid-Piacenzian climate using the LMDz atmospheric general circulation model (AGCM with a stretched grid which allows a finer resolution over Europe. In a first step, we applied the PRISM2 (Pliocene Research, Interpretation, and Synoptic Mapping boundary conditions except that we used modern terrestrial vegetation. Second, we simulated the vegetation for this period by forcing the ORCHIDEE (Organizing Carbon and Hydrology in Dynamic Ecosystems dynamic global vegetation model (DGVM with the climatic outputs from the AGCM. We then supplied this simulated terrestrial vegetation cover as an additional boundary condition in a second AGCM run. This gives us the opportunity to investigate the model's sensitivity to the simulated vegetation changes in a global warming context.

    Model results and data show a great consistency for mean annual temperatures, indicating increases by up to 4°C in the study area, and some disparities, in particular in the northern Mediterranean sector, as regards winter and summer temperatures. Similar continental mean annual precipitation and moisture patterns are predicted by the model, which broadly underestimates the wetter conditions indicated by the data in northwestern Europe. The biogeophysical effects due to the changes in vegetation simulated by ORCHIDEE are weak

  14. Enzyme assays.

    Science.gov (United States)

    Reymond, Jean-Louis; Fluxà, Viviana S; Maillard, Noélie

    2009-01-07

    Enzyme assays are analytical tools to visualize enzyme activities. In recent years a large variety of enzyme assays have been developed to assist the discovery and optimization of industrial enzymes, in particular for "white biotechnology" where selective enzymes are used with great success for economically viable, mild and environmentally benign production processes. The present article highlights the aspects of fluorogenic and chromogenic substrates, sensors, and enzyme fingerprinting, which are our particular areas of interest.

  15. Validation of turbulence and convective schemes on western Africa; comparison of LAM and CRM simulations on an AMMA case study.

    Science.gov (United States)

    Pollack, David; Gueremy, Jean-Francois; Beau, Isabelle

    2010-05-01

    The aim of this work is to analyse the behaviour of turbulence and convective parameterizations included in the Météo-France ALADIN-CLIMAT Limited Area Model in the frame of a 48 hour simulation of an AMMA case study, in comparison to observations and to a CRM (Méso-NH, with a 5 km horizontal grid-mesh) simulation carried out under the same boundary forcings. This framework provides an intermediate step of parameterization validation between the Single Column Model and Global Climate Model simulation studies. The chosen case study is the 26-27th July 2006 over a 43° x 40° region centred over Burkina Faso, in continuation to a previous work done with a HAPEX-Sahel case-study. During this 2 day period, two successive mesoscale convective systems are located ahead and in phase with the trough of an African easterly Wave (AEW). Both LAM and CRM simulations have been performed over the same considered domain, using the same ECMWF boundary forcings. Sensitivity tests to resolution (both horizontal and vertical) have been first carried out with ALADIN-CLIMAT. Second, the two different convection schemes used in ALADIN-CLIMAT show two kinds of response mainly due to their different formulations of triggering (no constraints in the dry layer under the convective cloud versus continuous treatment of convection including this dry layer) and closure (moisture convergence versus CAPE). Third, the impact of convective downdrafts will be shown. Fourth, the impact of different boundary forcing fields will also be presented.

  16. Evaluation of in vitro screening system for estrogenicity: comparison of stably transfected human estrogen receptor-α transcriptional activation (OECD TG455) assay and estrogen receptor (ER) binding assay.

    Science.gov (United States)

    Lee, Hae Kyung; Kim, Tae Sung; Kim, Chang Yeong; Kang, Il Hyun; Kim, Mi Gyeong; Jung, Ki Kyung; Kim, Hyung Sik; Han, Soon Young; Yoon, Hae Jung; Rhee, Gyu Seek

    2012-01-01

    The estrogenic activity of industrial chemicals, di(2-ethylhexyl) phthalate (DEHP), di(n-butyl) phthalate (DBP), benzylbutyl phthalate (BBP), diethyl phthalate (DEP), tetrabromobisphenol A (TBBPA), bisphenol A (BPA), and nonylphenol (NP), was compared using OECD test guideline 455(TG455), stably transfected transcriptional activation (STTA) and estrogen receptor (ER) binding assays. The estrogenic activity of BBP, BPA and NP were approximately 180,000-fold (PC(50), 4.32 x 10(-6 )M), 5,000-fold (PC(50), 1.26 x 10(-7) M) and 120,000-fold (PC(50), 2.92 x 10(-6 )M) less than 17β-estradiol (PC(50), 2.43 x 10(-11)M), whereas DEHP, DBP and DEP did not show any estrogenicity activity in the STTA assay. Moreover, binding affinities to human ERα of BBP, BPA, and NP were approximately 200,000-fold (IC(50), 4.91 x 10(-4) M), 8000-fold (IC(50), 1.92 x 10(-5) M) and 1400-fold (IC(50), 3.34 x 10(-6) M) less than 17β-estradiol (IC(50), 2.45 x 10(-9) M) in competitive human ERα binding assay. The relative potencies of STTA assay were very similar to ER binding, E-screen, and Yeast screening assays. Therefore, our results suggested that OECD test guideline TG455 may be useful as a screening test for potential endocrine disruptors.

  17. Detection of Escherichia coli O157 in raw and cooked meat: comparison of conventional direct culture method and Enzyme Linked Fluorescent Assay (ELFA

    Directory of Open Access Journals (Sweden)

    Maria De Giusti

    2011-03-01

    Full Text Available

    Abstract
    Background: Verocytotoxin Escherichia coli is a frequent and important cause of diarrhea and haemolytic uremic syndrome all over the world. Consumption of ground beef, lettuce, and other kinds of food have been associated with outbreaks.
    The aim of this study was to detect the presence of E. coli O157 in meat products collected from hospital food catering services in Rome, using a rapid detection method in comparison with a standard culture method to verify the effectiveness of HACCP system.
    Methods: Three hundred and ten food samples (80 of cooked and 230 of raw meat were screened for E.coli O157 by ISO culture method and by enzyme-linked-fluorescent-assay (ELFA-based methods
    (VIDAS®system, bioMérieux. All isolates obtained were tested for VT1 and VT2 genes by PCR. The statistical analysis considered absolute frequencies and percentages. The K statistic was applied to assess agreement between direct culture method and the VIDAS system.
    Results: A total of 6 (1,9% E.coli O157 isolates were recovered from raw meat samples by the culture method; of these only four were identified by PCR as VTEC producers. A total of 9 (2,9% E.coli O157 isolates were recovered from raw meat samples by the VIDAS® system. No E.coli O157 was detected in cooked products. All comparisons between the direct culture method and the VIDAS system were
    statistically significant (K= 0,795; p<0.001.
    Conclusions: The present study showed how ELFA-based methods are highly specific and rapid for the detection of E.coli O157 in food samples compared with the direct culture method. ELFA method is useful to verify the effectiveness of the HACCP system in the risk management of potential contaminating hazards during the preparation of foods for susceptible persons.

  18. Comparison of enzyme-linked immunosorbent assay and gas chromatography procedures for the detection of cyanazine and metolachlor in surface water samples

    Science.gov (United States)

    Schraer, S.M.; Shaw, D.R.; Boyette, M.; Coupe, R.H.; Thurman, E.M.

    2000-01-01

    Enzyme-linked immunosorbent assay (ELISA) data from surface water reconnaissance were compared to data from samples analyzed by gas chromatography for the pesticide residues cyanazine (2-[[4-chloro-6-(ethylamino)-l,3,5-triazin-2-yl]amino]-2-methylpropanenitrile ) and metolachlor (2-chloro-N-(2-ethyl-6-methylphenyl)-N-(2-methoxy-1-methylethyl)acetamide). When ELISA analyses were duplicated, cyanazine and metolachlor detection was found to have highly reproducible results; adjusted R2s were 0.97 and 0.94, respectively. When ELISA results for cyanazine were regressed against gas chromatography results, the models effectively predicted cyanazine concentrations from ELISA analyses (adjusted R2s ranging from 0.76 to 0.81). The intercepts and slopes for these models were not different from 0 and 1, respectively. This indicates that cyanazine analysis by ELISA is expected to give the same results as analysis by gas chromatography. However, regressing ELISA analyses for metolachlor against gas chromatography data provided more variable results (adjusted R2s ranged from 0.67 to 0.94). Regression models for metolachlor analyses had two of three intercepts that were not different from 0. Slopes for all metolachlor regression models were significantly different from 1. This indicates that as metolachlor concentrations increase, ELISA will over- or under-estimate metolachlor concentration, depending on the method of comparison. ELISA can be effectively used to detect cyanazine and metolachlor in surface water samples. However, when detections of metolachlor have significant consequences or implications it may be necessary to use other analytical methods.

  19. The comet assay with multiple mouse organs: comparison of comet assay results and carcinogenicity with 208 chemicals selected from the IARC monographs and U.S. NTP Carcinogenicity Database.

    Science.gov (United States)

    Sasaki, Y F; Sekihashi, K; Izumiyama, F; Nishidate, E; Saga, A; Ishida, K; Tsuda, S

    2000-11-01

    The comet assay is a microgel electrophoresis technique for detecting DNA damage at the level of the single cell. When this technique is applied to detect genotoxicity in experimental animals, the most important advantage is that DNA lesions can be measured in any organ, regardless of the extent of mitotic activity. The purpose of this article is to summarize the in vivo genotoxicity in eight organs of the mouse of 208 chemicals selected from International Agency for Research on Cancer (IARC) Groups 1, 2A, 2B, 3, and 4, and from the U.S. National Toxicology Program (NTP) Carcinogenicity Database, and to discuss the utility of the comet assay in genetic toxicology. Alkylating agents, amides, aromatic amines, azo compounds, cyclic nitro compounds, hydrazines, halides having reactive halogens, and polycyclic aromatic hydrocarbons were chemicals showing high positive effects in this assay. The responses detected reflected the ability of this assay to detect the fragmentation of DNA molecules produced by DNA single strand breaks induced chemically and those derived from alkali-labile sites developed from alkylated bases and bulky base adducts. The mouse or rat organs exhibiting increased levels of DNA damage were not necessarily the target organs for carcinogenicity. It was rare, in contrast, for the target organs not to show DNA damage. Therefore, organ-specific genotoxicity was necessary but not sufficient for the prediction of organ-specific carcinogenicity. It would be expected that DNA crosslinkers would be difficult to detect by this assay, because of the resulting inhibition of DNA unwinding. The proportion of 10 DNA crosslinkers that was positive, however, was high in the gastrointestinal mucosa, stomach, and colon, but less than 50% in the liver and lung. It was interesting that the genotoxicity of DNA crosslinkers could be detected in the gastrointestinal organs even though the agents were administered intraperitoneally. Chemical carcinogens can be classified

  20. Ectomycorrhizal fungi associated with ponderosa pine and Douglas-fir: a comparison of species richness in native western North American forests and Patagonian plantations from Argentina.

    Science.gov (United States)

    Barroetaveña, C; Cázares, E; Rajchenberg, M

    2007-07-01

    The putative ectomycorrhizal fungal species registered from sporocarps associated with ponderosa pine and Douglas-fir forests in their natural range distribution (i.e., western Canada, USA, and Mexico) and from plantations in south Argentina and other parts of the world are listed. One hundred and fifty seven taxa are reported for native ponderosa pine forests and 514 taxa for native Douglas-fir forests based on available literature and databases. A small group of genera comprises a high proportion of the species richness for native Douglas-fir (i.e., Cortinarius, Inocybe, and Russula), whereas in native ponderosa pine, the species richness is more evenly distributed among several genera. The comparison between ectomycorrhizal species richness associated with both trees in native forests and in Patagonia (Argentina) shows far fewer species in the latter, with 18 taxa for the ponderosa pine and 15 for the Douglas-fir. Epigeous species richness is clearly dominant in native Douglas-fir, whereas a more balanced relation epigeous/hypogeous richness is observed for native ponderosa pine; a similar trend was observed for Patagonian plantations. Most fungi in Patagonian Douglas-fir plantations have not been recorded in plantations elsewhere, except Suillus lakei and Thelephora terrestris, and only 56% of the fungal taxa recorded in Douglas-fir plantations around the world are known from native forests, the other taxa being new associations for this host, suggesting that new tree + ectomycorrhizal fungal taxa associations are favored in artificial situations as plantations.

  1. 人文视野下的中西方管理思想比较%Comparison of Management Ideas in China and Western Countries From Humanistic Perspective

    Institute of Scientific and Technical Information of China (English)

    章迪诚; 张星伍

    2012-01-01

    在承认管理理论具有超越地域文化的普适性基础上,以中西方管理思想的比较为逻辑起点,通过比较中西方在管理视角、管理基点、管理方法、管理行为和管理原则等方面存在诸多的差异,力图为管理理论在中国的学科建设中建立不同于西方的测量维度,提供可资观察和应用的理论工具.%Agreeing with the fact that management is certainly beyond geographical and cultural content of the universal basis, this article analyzes the differences between management in China and west countries from the aspects of managerial perspectives, foundations, methods, behaviors and principles based upon managerial idea comparison. It is expected to provide theoretical tools for establishing measurement dimensions for management discipline development, which are different from those in western countries.

  2. The importance of forest floor disturbance in the early regeneration patterns of the boreal forest of western and central Quebec : a wildfire versus logging comparison

    Energy Technology Data Exchange (ETDEWEB)

    Nguyen-Xuan, T.; Bergeron, Y.; Fyles, J.W.; Simard, D.; Pare, D. [Quebec Univ., Montreal, PQ (Canada) Dept. de sciences biologiques

    2000-09-01

    The different forestry practices of wildfire and logging were compared to better understand their capability to address sustainable forest management issues. Three separate areas of the black spruce feathermoss forest of western and central Quebec were studied. The comparison entailed nonvascular and vascular plant composition of the early regenerating vegetation present after either clear-cut logging or wildfire. Along the first ordination axis, the authors used a detrended correspondence analysis to differentiate the burned and logged stands in each area. There was a greater abundance of pioneer species or lichens following a fire and a greater abundance of residual species following clear-cutting. In order to relate variables that characterized the physical disturbance of the forest floor and the general site conditions to the two first differentiating axes, the Spearman's correlation coefficients were calculated. In two of the three study areas, the variables that characterized forest floor disturbance severity were strongly associated with the first ordination axis. The identification of regeneration patterns was made possible by the interpretation of compositional differences in light of plant reproductive strategies. It described the influence of disturbance type and severity on post-disturbance vegetation composition. The results seemed to indicate that select forestry practices like careful logging with the protection of regeneration and soil, scarification, and prescribed burning possess different capabilities with regard to sustainable forest management practices. 56 refs., 4 tabs., 3 figs.

  3. Increasing vegetation and climate gradient in Western Europe over the Last Glacial Inception (122 110 ka): data-model comparison

    Science.gov (United States)

    Sánchez Goñi, M. F.; Loutre, M. F.; Crucifix, M.; Peyron, O.; Santos, L.; Duprat, J.; Malaizé, B.; Turon, J.-L.; Peypouquet, J.-P.

    2005-02-01

    vegetation and climate gradients in northeastern Atlantic and European borderlands probably related with the well-developed ice caps at that time. The comparison between the general trend in the estimated and simulated MoBidiC winter and summer temperatures for latitudes between 35 and 45° N, shows that both follow quite straightforwardly the precession signal although the simulated and reconstructed temperatures agree better in the South than North of 40° N. Annual precipitation is exhibiting opposite trend in the data and in the model. This contradiction is likely the fact that the zonal climate simulated by the model may not accurately represent the regional climate features, as reconstructed from the pollen.

  4. The comparison between two airborne LiDAR datasets to analyse debris flow initiation in north-western Iceland

    Science.gov (United States)

    Morino, Costanza; Conway, Susan J.; Balme, Matthew R.; Jordan, Colm; Hillier, John; Sæmundsson, Þorsteinn; Argles, Tom

    2015-04-01

    A debris flow is a very rapid to extremely rapid flow (e.g., 0.8-28 ms-1) [1], that occurs when coarse and poorly-sorted debris, mixed with water and/or air, move down hill slopes in response to gravity [2]. Both the fluid and the solid have a strong influence on the movement of debris flows. They can be extremely destructive, due to their capability of transporting metre-size boulders [e.g., 3, 4]. There are two main ways in which a debris flow can be initiated: by slope failure or by the "fire hose" effect. The slope failure type is particularly common in alpine regions, where landslides can evolve into debris flows [5], triggered by the coalescence of different slope failures. Steep slope gradients, high pore-water pressures, heavy rainfall and/or snowmelt favour this process. The "fire hose" effect occurs when there is a high concentration of debris accumulated within a pre-existing channel; a surge of water through the channel can then develop into a debris flow by incorporating this debris [e.g. 5-7]. In this study, we examine the triggering style of debris flows above the town of Ísafjörður in the Westfjords of Iceland. The slope above the town is characterised by a large topographic bench upon which 20-35 m of glacial till is perched. The sediments are unstable at the bench margin and thus generate frequent, large, hillslope debris flows [8, 9]. In our new analysis, we report on the comparison between the two airborne LiDAR elevation models (collected in 2007 and 2013 by the UK Natural Environment Research Council Airborne Research and Survey Facility), which display several new debris flows and also related mass movements. From these analyses, we find that debris flows in the region are triggered by simple failure of the glacial till, as recognised before [8, 9]. However, debris flows may also be regenerated by the "fire hose" effect, when debris that has collapsed into chutes is remobilised by a later snowmelt or precipitation event. Comparing

  5. Comparison between a Broad-Range Real-Time and a Broad-Range End-Point PCR Assays for the Detection of Bacterial 16S rRNA in Clinical Samples.

    Science.gov (United States)

    Meddeb, Mariam; Koebel, Christelle; Jaulhac, Benoît; Schramm, Frédéric

    2016-01-01

    Broad range PCR targeting the 16S rRNA gene is widely used to test clinical samples for the presence of bacterial DNA. End-point 16S PCR is both time-consuming and at high risk of cross-contamination. Prior to the replacement of the 16S end-point PCR assay routinely used in our clinical laboratory by a new 16S real-time PCR assay, we aimed to compare the performances of both techniques for the direct diagnosis of bacterial infections in clinical samples. In this prospective study, 129 clinical samples were included for direct comparison of both techniques. The sensitivity of 16S real-time PCR assay (76%) was significantly higher than that of end-point 16S PCR assay (41%) (pPCR assays did not differ significantly (p=0.43). The 16S real-time PCR assay yielded an etiological diagnosis in 19% of culture-negative samples. It constitutes a reliable and complementary diagnostic tool to the bacterial culture.

  6. Evaluation of the Binax NOW Flu A+B Enzyme Immunochromatographic Assay in comparison with Real-Time PCR during the Pandemic of Influenza 2009

    Directory of Open Access Journals (Sweden)

    Iris Hasibra (Hatibi

    2013-12-01

    Full Text Available The Binax NOW Flu A+B enzyme immunochromatographic assay was compared to Real-Time PCR assay for 542 specimen from nasal-wash or nasopharyngeal swab collected during the pandemic of 2009. The overall sensitivity, specificity, positive predic¬tive value, and negative predictive value of the assay were 44.6%, 95.8%, 73.5%, and 86.9%, respectively. The assay sensitivity shows mixed values decreasing significantly in infants and children age, which is linked with the quality and the way sample is collected.

  7. Comparison of Assays for Sensitive and Reproducible Detection of Cell Culture-Infectious Cryptosporidium parvum and Cryptosporidium hominis in Drinking Water

    Science.gov (United States)

    Di Giovanni, George D.; Rochelle, Paul A.

    2012-01-01

    This study compared the three most commonly used assays for detecting Cryptosporidium sp. infections in cell culture: immunofluorescent antibody and microscopy assay (IFA), PCR targeting Cryptosporidium sp.-specific DNA, and reverse transcriptase PCR (RT-PCR) targeting Cryptosporidium sp.-specific mRNA. Monolayers of HCT-8 cells, grown in 8-well chamber slides or 96-well plates, were inoculated with a variety of viable and inactivated oocysts to assess assay performance. All assays detected infection with low doses of flow cytometry-enumerated Cryptosporidium parvum oocysts, including infection with one oocyst and three oocysts. All methods also detected infection with Cryptosporidium hominis. The RT-PCR assay, IFA, and PCR assay detected infection in 23%, 25%, and 51% of monolayers inoculated with three C. parvum oocysts and 10%, 9%, and 16% of monolayers inoculated with one oocyst, respectively. The PCR assay was the most sensitive, but it had the highest frequency of false positives with mock-infected cells and inactivated oocysts. IFA was the only infection detection assay that did not produce false positives with mock-infected monolayers. IFA was also the only assay that detected infections in all experiments with spiked oocysts recovered from Envirochek capsules following filtration of 1,000 liters of treated water. Consequently, cell culture with IFA detection is the most appropriate method for routine and sensitive detection of infectious Cryptosporidium parvum and Cryptosporidium hominis in drinking water. PMID:22038611

  8. Comparison of peptide nucleic acid fluorescence in situ hybridization assays with culture-based matrix-assisted laser desorption/ionization-time of flight mass spectrometry for the identification of bacteria and yeasts from blood cultures and cerebrospinal fluid cultures.

    Science.gov (United States)

    Calderaro, A; Martinelli, M; Motta, F; Larini, S; Arcangeletti, M C; Medici, M C; Chezzi, C; De Conto, F

    2014-08-01

    Peptide nucleic acid fluorescence in situ hybridization (PNA FISH) is a molecular diagnostic tool for the rapid detection of pathogens directly from liquid media. The aim of this study was to prospectively evaluate PNA FISH assays in comparison with culture-based matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) identification, as a reference method, for both blood and cerebrospinal fluid (CSF) cultures, during a 1-year investigation. On the basis of the Gram stain microscopy results, four different PNA FISH commercially available assays were used ('Staphylococcus aureus/CNS', 'Enterococcus faecalis/OE', 'GNR Traffic Light' and 'Yeasts Traffic Light' PNA FISH assays, AdvanDx). The four PNA FISH assays were applied to 956 positive blood cultures (921 for bacteria and 35 for yeasts) and 11 CSF cultures. Among the 921 blood samples positive for bacteria, PNA FISH gave concordant results with MALDI-TOF MS in 908/921 (98.64%) samples, showing an agreement of 99.4% in the case of monomicrobial infections. As regards yeasts, the PNA FISH assay showed a 100% agreement with the result obtained by MALDI-TOF MS. When PNA FISH assays were tested on the 11 CSF cultures, the results agreed with the reference method in all cases (100%). PNA FISH assays provided species identification at least one work-day before the MALDI-TOF MS culture-based identification. PNA FISH assays showed an excellent efficacy in the prompt identification of main pathogens, yielding a significant reduction in reporting time and leading to more appropriate patient management and therapy in cases of sepsis and severe infections.

  9. Comparison of PCR-ELISA and LightCycler real-time PCR assays for detecting Salmonella spp. in milk and meat samples

    DEFF Research Database (Denmark)

    Perelle, Sylvie; Dilasser, Françoise; Malorny, Burkhard

    2004-01-01

    , minced beef and raw milk, and 92 naturally-contaminated milk and meat samples. When using either PCR-ELISA or LC-PCR assays, only Salmonella strains were detected. PCR-ELISA and LC-PCR assays gave with pure Salmonella cultures the same detection limit level of 10(3) CFU/ml, which corresponds respectively...

  10. Validation and comparison of two commercial ELISA kits and three in-house developed real-time PCR assays for the detection of potentially allergenic mustard in food.

    Science.gov (United States)

    Palle-Reisch, Monika; Hochegger, Rupert; Štumr, Stepan; Korycanova, Kveta; Cichna-Markl, Margit

    2015-05-01

    The study compares the applicability of two commercial mustard ELISA kits (Mustard ELISA Kit-specific and Mustard ELISA Kit-total) and three in-house developed real-time PCR assays (singleplex assay for white mustard, singleplex assay for black/brown mustard and duplex assay for the detection of white, black and brown mustard). Analyses of raw and brewed model sausages containing white and black/brown mustard in the range from 1 to 50 ppm indicate that both ELISAs and the three real-time PCR assays allow the detection of traces of mustard in raw and in brewed sausages. The ELISAs were found to be more sensitive than the real-time PCR assays. When the ELISAs and real-time PCR assays were applied to the analysis of 15 commercial foodstuffs differing in their labelling concerning mustard, in one sample mustard was detected with both ELISAs and the three real-time PCR assays although mustard was not indicated on the food ingredient list.

  11. Use of an improved E. coli method for the measurement of cobalamin in serum: comparison with the E. gracilis assay results.

    Science.gov (United States)

    Sourial, N A

    1981-01-01

    Owing to the higher serum cobalamin results that are obtained by R-binder radioisotopic dilution assay compared to microbiological assays (E. gracilis and L. leichmannii) it was suggested that serum contained a cobamide(s) that could not be detected by the more specific microbiological assays and that a much less specific test organism, which responds to most naturally occurring cobamides, such as the cobamide-dependent E. coli mutant, might respond to these cobamide(s) in serum. In an attempt to investigate this possibility an improved and simplified E. coli assay for the measurement of cobamide in serum was developed. The method is described, and the results obtained in normal subjects, in patients with megaloblastic anemia, and in anaemic pregnant women not suffering from megaloblastic anaemia are reported and compared with E. gracilis assay results. PMID:6787097

  12. Comparison of cardiac TnI outliers using a contemporary and a high-sensitivity assay on the Abbott Architect platform.

    Science.gov (United States)

    Ryan, J B; Southby, S J; Stuart, L A; Mackay, R; Florkowski, C M; George, P M

    2014-07-01

    Assays for cardiac troponin (cTn) have undergone improvements in sensitivity and precision in recent years. Increased rates of outliers, however, have been reported on various cTn platforms, typically giving irreproducible, falsely higher results. We aimed to evaluate the outlier rate occurring in patients with elevated cTnI using a contemporary and high-sensitivity assay. All patients with elevated cTnI (up to 300 ng/L) performed over a 21-month period were assayed in duplicate. A contemporary assay (Abbott STAT Troponin-I) was used for the first part of the study and subsequently a high-sensitivity assay (Abbott STAT High-Sensitive Troponin-I) was used. Outliers exceeded a calculated critical difference (CD) (CD = z × √2 × SDAnalytical) where z = 3.5 (for probability of 0.0005) and critical outliers also were on a different side of the decision level. The respective outlier and critical outlier rates were 0.22% and 0.10% for the contemporary assay (n = 4009) and 0.18% and 0.13% for the high-sensitivity assay (n = 3878). There was no significant reduction in outlier rate between the two assays (χ(2) = 0.034, P = 0.854). Fifty-six percent of outliers occurred in samples where cTn was an 'add-on' test (and was stored and refrigerated prior to assay). Despite recent improvements in cTn methods, outliers (including critical outliers) still occur at a low rate in both a contemporary and high-sensitivity cTnI assay. Laboratory and clinical staff should be aware of this potential analytical error, particularly in samples with suboptimal sample handling such as add-on tests. © The Author(s) 2014 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.

  13. Comparison of an In-house and a Commercial RD1-based ELISPOT-IFN-γ Assay for the Diagnosis of Mycobacterium tuberculosis Infection

    Science.gov (United States)

    Mantegani, Paola; Piana, Federica; Codecasa, Luigi; Galli, Laura; Scarpellini, Paolo; Lazzarin, Adriano; Cirillo, Daniela; Fortis, Claudio

    2006-01-01

    Objective: To compare a RD1-based in-house ELISPOT-interferon-γ (IFN-γ) assay with a commercial (T-SPOT.TB™) assay for the diagnosis of Mycobacterium tuberculosis (TB) infection and the efficacy of the tuberculin skin test (TST) and ELISPOT assay in detecting latent TB infection (LTBI). Design: Eighty-six subjects (65 household contacts of contagious TB-infected patients, 13 subjects with active or previous TB infection, and 8 with suspected TB infection) were consecutively recruited in the context of a surveillance program. Methods: Enrolled subjects underwent the Mantoux TST and two different ELISPOT-IFN-γ assays: an in-house assay using a pool of selected M. tuberculosis peptides (MTP) and the commercial T-SPOT.TB assay. Results: The in-house and commercial ELISPOT-IFN-γ assays showed almost complete concordance (99%) in diagnosing acute or LTBI.When comparing the efficacy of the TST with the in-house ELISPOT assay in detecting TB infection, a small agreement was observed (k=0.344, P<0.0001): 36% of the subjects with a positive TST were ELISPOT-MTP negative and 12% with a negative TST were ELISPOT-MTP positive. Furthermore, 78% of the ELISPOT-MTP negative individuals were ELISPOT- Bacillus Calmette-Guérin (BCG) positive, most of whom had received BCG vaccination. Conclusion: Our in-house ELISPOT assay based on a restricted pool of highly selected peptides is equivalent to the commercial T-SPOT.TB assay, is cheaper and is probably not confounded, unlike the TST, by BCG vaccination in our setting PMID:17210976

  14. Peri-operative troponin monitoring using a prototype high-sensitivity cardiac troponin I (hs-cTnI) assay: comparisons with hs-cTnT and contemporary cTnI assays.

    LENUS (Irish Health Repository)

    Lee, Graham R

    2013-09-18

    Non-cardiac surgery is associated with major vascular complications and higher incidences of elevated plasma troponin (cTn) concentration. Goal-directed therapy (GDT) is a stroke volume (SV)-guided approach to intravenous (IV) fluid therapy that improves tissue perfusion, oxygenation and reduces post-operative complications. In patients undergoing major gastro-intestinal surgery, we compared high sensitive and contemporary troponin assays and correlated results with patient outcome.

  15. Detection of anti-infliximab antibodies is impacted by antibody titer, infliximab level and IgG4 antibodies: a systematic comparison of three different assays

    Science.gov (United States)

    Afonso, Joana; Lopes, Susana; Gonçalves, Raquel; Caldeira, Paulo; Lago, Paula; Tavares de Sousa, Helena; Ramos, Jaime; Gonçalves, Ana Rita; Ministro, Paula; Rosa, Isadora; Vieira, Ana Isabel; Coelho, Rosa; Tavares, Patrícia; Soares, João; Sousa, Ana Lúcia; Carvalho, Diana; Sousa, Paula; da Silva, João Pereira; Meira, Tânia; Silva Ferreira, Filipa; Dias, Cláudia Camila; Chowers, Yehuda; Ben-Horin, Shomron; Magro, Fernando

    2016-01-01

    Background: There is scant information on the accuracy of different assays used to measure anti-infliximab antibodies (ADAs), especially in the presence of detectable infliximab (IFX). We thus aimed to evaluate and compare three different assays for the detection of IFX and ADAs and to clarify the impact of the presence of circulating IFX on the accuracy of the ADA assays. Methods: Blood samples from 79 ulcerative colitis (UC) patients treated with infliximab were assessed for IFX levels and ADAs using three different assays: an in-house assay and two commercial kits, Immundiagnostik and Theradiag. Sera samples with ADAs and undetectable levels of IFX were spiked with exogenous IFX and analyzed for ADAs. Results: The three assays showed 81–96% agreement for the measured IFX level. However, the in-house assay and Immundiagnostik assays detected ADAs in 34 out of 79 samples, whereas Theradiag only detected ADAs in 24 samples. Samples negative for ADAs with Theradiag, but ADA-positive in both the in-house and Immundiagnostik assays, were positive for IFX or IgG4 ADAs. In spiking experiments, a low concentration of exogenous IFX (5 µg/ml) hampered ADA detection with Theradiag in sera samples with ADA levels of between 3 and 10 µg/ml. In the Immundiagnostik assay detection interference was only observed at concentrations of exogenous IFX higher than 30 µg/ml. However, in samples with high levels of ADAs (>25 µg/ml) interference was only observed at IFX concentrations higher than 100 µg/ml in all three assays. Binary (IFX/ADA) stratification of the results showed that IFX+/ADA- and IFX-/ADAs+ were less influenced by the assay results than the double-positive (IFX+/ADAs+) and double-negative (IFX-/ADAs-) combination. Conclusions: All three methodologies are equally suitable for measuring IFX levels. However, erroneous therapeutic decisions may occur when patients show double-negative (IFX-/ADAs-) or double-positive (IFX+/ADAs+) status, since agreement between

  16. Comparison of five different in vitro assays for assessment of sodium metavanadate cytotoxicity in Chinese hamster ovary cells (CHO-K1 line).

    Science.gov (United States)

    Zwolak, Iwona

    2015-08-01

    This investigation was undertaken to compare five different in vitro cytotoxicity assays for their power in revealing vanadium-mediated toxicity in Chinese hamster ovary (CHO)-K1 cells. The cells were exposed to sodium metavanadate (NaVO(3)) in the range of 10-1000 µM for 24 h and thereafter the cytotoxic effects of NaVO(3) were measured by colorimetric in vitro assays: the neutral red (NR) test, the 2,3-bis[2-methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxyanilide inner salt (XTT) assay, the resazurin assay, the sulforhodamine B (SR-B) assay, and by microscopic assessment of cell viability using the trypan blue (TB) staining method. Among the assays used, the NR test was the most sensitive, since it revealed metavanadate cytotoxicity at the lowest NaVO(3) dose (=50 µM). Also, NaVO(3) cytotoxicity expressed as inhibitory concentration (IC) showed the lowest values for the NR test. Three other tests XTT, resazurin, and SR-B assays showed intermediate sensitivity revealing the cytotoxicity of NaVO(3) at 100 µM. The corresponding IC10 and IC50 values calculated for the XTT, resazurin, and SR-B tests were similar. The TB staining method was the least sensitive, since it recorded metavanadate cytotoxicity at the highest NaVO(3) concentration tested (=600 µM). Based on the cytotoxicity end points measured with the above assays, it can be concluded that lysosomal/Golgi apparatus damage (measured by NR assay) may be the primary effect of NaVO(3) on CHO-K1 cells. The disintegration of mitochondria (assessed with the XTT and resazurin assays) probably follows lysosomal impairment. Plasma membrane permeability (staining with TB) occurs at a late stage of NaVO(3)-induced cytotoxicity on CHO-K1 cells. The results obtained in this research work show that the NR test can be recommended as a very sensitive assay for the assessment of NaVO(3) cytotoxicity in the CHO-K1 cell culture model. Considering the convenience of assay performance along with adequate sensitivity

  17. Evaluation of Estrogenic Activity of Wastewater: Comparison Among In Vitro ERα Reporter Gene Assay, In Vivo Vitellogenin Induction, and Chemical Analysis.

    Science.gov (United States)

    Ihara, Masaru; Kitamura, Tomokazu; Kumar, Vimal; Park, Chang-Beom; Ihara, Mariko O; Lee, Sang-Jung; Yamashita, Naoyuki; Miyagawa, Shinichi; Iguchi, Taisen; Okamoto, Seiichiro; Suzuki, Yutaka; Tanaka, Hiroaki

    2015-05-19

    The in vitro estrogen receptor (ER) reporter gene assay has long been used to measure estrogenic activity in wastewater. In a previous study, we demonstrated that the assay represents net estrogenic activity in the balance between estrogenic and antiestrogenic activities in wastewater. However, it remained unclear whether the net estrogenic activity measured by the in vitro ERα reporter gene assay can predict the in vivo estrogenic effect of wastewater. To determine this, we measured the following: estrogenic and antiestrogenic activities of wastewater and reclaimed water by the in vitro ERα reporter gene assay, expression of vitellogenin-1 (vtg1) and choriogenin-H (chgH) in male medaka (Oryzias latipes) by quantitative real-time PCR, and estrone, 17β-estradiol, estriol, and 17α-ethynylestradiol concentrations chemically to predict estrogenic activity. The net estrogenic activity measured by the in vitro medaka ERα reporter gene assay predicted the in vivo vtg1/chgH expression in male medaka more accurately than the concentrations of estrogens. These results also mean that in vivo vtg1/chgH expression in male medaka is determined by the balance between estrogenic and antiestrogenic activities. The in vitro medaka ERα reporter gene assay also predicted in vivo vtg1/chgH expression on male medaka better than the human ERα reporter gene assay.

  18. Comparison of the performance in detection of HPV infections between the high-risk HPV genotyping real time PCR and the PCR-reverse dot blot assays.

    Science.gov (United States)

    Zhang, Lahong; Dai, Yibei; Chen, Jiahuan; Hong, Liquan; Liu, Yuhua; Ke, Qiang; Chen, Yiwen; Cai, Chengsong; Liu, Xia; Chen, Zhaojun

    2017-08-29

    A new multiplex real-time PCR assay, the high-risk HPV genotyping real time PCR assay (HR HPV RT-PCR), has been developed to detect 15 high-risk HPV types with respective viral loads. In this report, a total of 684 cervical specimens from women diagnosed with vaginitis were assessed by the HR HPV RT-PCR and the PCR reaction and reverse dot blot (PCR-RDB) assays, using a PCR-sequencing method as a reference standard. A total coincidence of 97.7% between the HR HPV RT PCR and the PCR-RDB assays was determined with a Kappa value of 0.953. The HR HPV RT PCR assay had sensitivity, specificity, and concordance rates (accuracy) of 99.7%, 99.7%, and 99.7%, respectively, as confirmed by PCR-sequencing, while the PCR-RDB assay had respective rates of 98.8%, 97.1%, and 98.0%. The overall rate of HPV infection, determined by PCR-sequencing, in women diagnosed with vaginitis was 49.85%, including 36.26% of single infection and 13.6% of multiple infections. The most common infections among the 15 high-risk HPV types in women diagnosed with vaginitis were HPV-52, HPV-16, and HPV-58, with a total detection rate of 10.23%, 7.75%, and 5.85%, respectively. We conclude that the HR HPV RT PCR assay exhibits better clinical performance than the PCR-RDB assay, and is an ideal alternative method for HPV genotyping. In addition, the HR HPV RT PCR assay provides HPV DNA viral loads, and could serve as a quantitative marker in the diagnosis and treatment of single and multiple HPV infections. © 2017 Wiley Periodicals, Inc.

  19. Late Weichselian sediment geochemistry of the western Barents Sea margin - an empirical inter-instrumental comparison of core scanning and conventional XRF measurements

    Science.gov (United States)

    Klug, Martin; Knies, Jochen; Forwick, Matthias; Haflidason, Haflidi

    2014-05-01

    During the last years an increasing number of studies in geosciences made use of the fast and non-destructive XRF scanning method. To create robust and reproducible data and to interpret geochemical variations across records of different origin and from different instrumentations inter-instrumental comparison becomes a necessary, inevitable and decisive procedure. In this study we present results from an empirical approach of an inter-instrumental XRF comparison including the Avaatech (University of Tromsø), Itrax (University of Bergen) and InnovX-GeoTek (The Geological Survey of Norway) core scanners. In addition single samples were measured with the PANalytical AXIOS XRF spectrometer and the Perkin Elmer 4300 Dual View ICP-AES measurements (both at the Geological Survey of Norway). We analysed the split-surface of a 300 cm long marine sediment core from the continental slope of the western Barents Sea (71°30'N, 16°10' E). The sediment core sections were logged near-continuously with the core scanners along the centre of the core axis, followed by measurements of discrete samples. All devices were standardized and calibrated prior measurements according to the individual, requisite standardisation routines. Results presented here were harmonized to common sampling midpoints. We tested element ratios commonly used in geosciences. Most of the down-core variations of element ratios from the core scans occur in general synchronously and match the variability of single sample measurements from the stand-alone XRF-analyzer indicating a convenient XRF technique implementation in the scanning instruments. However, in certain cases, element ratios appear to show very low variations, likely an indication of detection-limit problems or larger uncertainties associated with the determination of low element concentrations. Apart from good relative fit, absolute variations occur at different levels and instrumental deviation varies for particular element ratios. This likely

  20. Development and comparison of a real-time PCR assay for detection of Dichelobacter nodosus with culturing and conventional PCR: harmonisation between three laboratories

    DEFF Research Database (Denmark)

    Frosth, Sara; Slettemeås, Jannice S.; Jørgensen, Hannah J.

    2012-01-01

    BACKGROUND: Ovine footrot is a contagious disease with worldwide occurrence in sheep. The main causative agent is the fastidious bacterium Dichelobacter nodosus. In Scandinavia, footrot was first diagnosed in Sweden in 2004 and later also in Norway and Denmark. Clinical examination of sheep feet...... a TaqMan-based real-time PCR assay for detection of D. nodosus and to compare its performance with culturing and conventional PCR. METHODS: A D. nodosus-specific TaqMan based real-time PCR assay targeting the 16S rRNA gene was designed. The inclusivity and exclusivity (specificity) of the assay...... was tested using 55 bacterial and two fungal strains. To evaluate the sensitivity and harmonisation of results between different laboratories, aliquots of a single DNA preparation were analysed at three Scandinavian laboratories. The developed real-time PCR assay was compared to culturing by analysing 126...

  1. Performance of the Aptima High-Risk Human Papillomavirus mRNA Assay in a Referral Population in Comparison with Hybrid Capture 2 and Cytology▿

    Science.gov (United States)

    Clad, Andreas; Reuschenbach, Miriam; Weinschenk, Johanna; Grote, Ruth; Rahmsdorf, Janina; Freudenberg, Nikolaus

    2011-01-01

    This study compared the Aptima human papillomavirus (HPV) (AHPV; Gen-Probe Incorporated) assay, which detects E6/E7 mRNA from 14 high-risk types, the Hybrid Capture 2 HPV DNA (HC2; Qiagen Incorporated) test, and repeat cytology for their ability to detect high-grade cervical lesions (cervical intraepithelial neoplasia grade 2+ [CIN2+]) in women referred to colposcopy due to an abnormal Papanicolaou (Pap) smear. A total of 424 clinical specimens, stored in liquid-based cytology (LBC) vials at room temperature for up to 3 years, were tested by repeat cytology, the AHPV assay, and the HC2 test. Assay results were compared to each other and to histology results. The overall agreement between the AHPV assay and the HC2 test was 88.4%. The sensitivity (specificity) of cytology, the HC2 test, and the AHPV assay for the detection of CIN2+ was 84.9% (66.3%), 91.3% (61.0%), and 91.7% (75.0%) and for the detection of CIN3+ was 93.9% (54.4%), 95.7% (46.0%), and 98.2% (56.3%), respectively. Of the disease-positive specimens containing high-risk HPV (HR HPV) DNA as determined by Linear Array (Roche Diagnostics), the AHPV assay missed 3 CIN2 and 1 microfocal CIN3 specimen, while the HC2 test missed 6 CIN2, 4 CIN3, and 1 cervical carcinoma specimen. The AHPV assay had a sensitivity similar to but a specificity significantly higher (P < 0.0001) than the HC2 test for the detection of CIN2+. The AHPV assay was significantly more sensitive (P = 0.0041) and significantly more specific (P = 0.0163) than cytology for the detection of disease (CIN2+). PMID:21191046

  2. Performance of the Aptima high-risk human papillomavirus mRNA assay in a referral population in comparison with Hybrid Capture 2 and cytology.

    Science.gov (United States)

    Clad, Andreas; Reuschenbach, Miriam; Weinschenk, Johanna; Grote, Ruth; Rahmsdorf, Janina; Freudenberg, Nikolaus

    2011-03-01

    This study compared the Aptima human papillomavirus (HPV) (AHPV; Gen-Probe Incorporated) assay, which detects E6/E7 mRNA from 14 high-risk types, the Hybrid Capture 2 HPV DNA (HC2; Qiagen Incorporated) test, and repeat cytology for their ability to detect high-grade cervical lesions (cervical intraepithelial neoplasia grade 2+ [CIN2+]) in women referred to colposcopy due to an abnormal Papanicolaou (Pap) smear. A total of 424 clinical specimens, stored in liquid-based cytology (LBC) vials at room temperature for up to 3 years, were tested by repeat cytology, the AHPV assay, and the HC2 test. Assay results were compared to each other and to histology results. The overall agreement between the AHPV assay and the HC2 test was 88.4%. The sensitivity (specificity) of cytology, the HC2 test, and the AHPV assay for the detection of CIN2+ was 84.9% (66.3%), 91.3% (61.0%), and 91.7% (75.0%) and for the detection of CIN3+ was 93.9% (54.4%), 95.7% (46.0%), and 98.2% (56.3%), respectively. Of the disease-positive specimens containing high-risk HPV (HR HPV) DNA as determined by Linear Array (Roche Diagnostics), the AHPV assay missed 3 CIN2 and 1 microfocal CIN3 specimen, while the HC2 test missed 6 CIN2, 4 CIN3, and 1 cervical carcinoma specimen. The AHPV assay had a sensitivity similar to but a specificity significantly higher (P < 0.0001) than the HC2 test for the detection of CIN2+. The AHPV assay was significantly more sensitive (P = 0.0041) and significantly more specific (P = 0.0163) than cytology for the detection of disease (CIN2+).

  3. [Comparison of the performance of the ECLusys anti-HCV reagent with the Lumipulse f and HISCL 2000-i HCVAb assays].

    Science.gov (United States)

    Sugiura, Aya; Iwahara, Kunihiro; Suga, Yasuyuki; Uchiyama, Sachinori; Maekawa, Masato

    2012-09-01

    We compared the ECLusys Anti-HCV (ECL) reagent to the Lumipulse f (LPf) and HISCL (HIS) HCV assays. In a correlation test using 210 routine clinical specimens measured using the Lumipulse method (96 positive and 114 negative), most of the results were consistent for all specimens. In a dilution sensitivity test using three different routine positive specimens, the ECL assay enabled detection at higher levels of sensitivity than either the LPf or the HIS assay. Moreover, when the distribution of the cut-off index (C.O.I.) values of the routine LPf negative specimens were compared to those on the ECL and HIS assays, it was found that on the ECL assay, most of the specimens had cut-off index values < 0.1, indicating a more clear-cut distribution. In a specificity test using high RF positive specimens(n = 33), pregnancy specimens (n = 35), cytomegalovirus (CMV) antibody positive specimens (n = 36), and high M protein positive specimens (n = 21), the ECL assay yielded positive results for a CMV antibody positive specimen and three high M protein positive specimens. Further testing using samples from the same patients collected on different days than these four samples resulted in a second positive result for the CMV positive specimen, and single antigen measurement yielded a Core/NS3 positive result, as well, suggesting past infection. However, since negative results were obtained for the three M protein positive specimens, the possibility of this being a ECLusys non-specific reaction could not be ruled out. The above results confirmed that the ECL assay provides superior fundamental performance, and possesses test performance nearly identical to that of the existing measurement methods that are widely used at a large number of facilities, and would therefore be a suitable assay for use in routine HCV antibody screening.

  4. Comparison of three stool antigen assays with the 13C- urea breath test for the primary diagnosis of Helicobacter pylori infection and monitoring treatment outcome.

    LENUS (Irish Health Repository)

    Hooton, Carmel

    2012-02-03

    BACKGROUND: The urea breath test (UBT) is the gold-standard non-invasive test for the detection of Helicobacter pylori infection, however, the lack of availability of the UBT due to the high cost of the test, and in particular the need for expensive analytical instrumentation, limits the usefulness of this method. Stool antigen assays may offer an alternative non-invasive method for the diagnosis of infection. OBJECTIVE: To compare the accuracy of three stool antigen assays (HpSA, IDEIA HpStAR, and ImmunoCard STAT) against the UBT for the primary diagnosis of H. pylori infection and for monitoring treatment outcome. METHODS: A total of 102 patients attending two gastroenterology day-case clinics for the investigation of dyspepsia were included. Each patient provided breath and stool samples for analysis. Patients who tested positive for H. pylori by the validated UBT were prescribed triple therapy and invited to return for repeat breath and stool sample analysis 6 weeks post-treatment. RESULTS: Of the 102 patients tested, 48 were diagnosed with H. pylori infection by the UBT. The HpSA assay interpreted 38 of these as positive (79% sensitive). Of the 54 UBT-negative patients the HpSA assay interpreted all 54 as negative (100% specific). The IDEIA HpStAR assay correctly identified 44 patients as positive (92% sensitive) and 50 as negative (92.5% specific). The ImmunoCard STAT assay interpreted 38 patients as positive (79% sensitive) and 52 as negative (96.3% specific). CONCLUSION: The findings indicate that the IDEIA HpStAR stool antigen kit is the most accurate assay of the three assays evaluated, and possibly represents a viable alternative to the UBT for the primary diagnosis of H. pylori infection and for monitoring treatment outcome.

  5. Quantitative multiplex real-time PCR assay for shrimp allergen: comparison of commercial master mixes and PCR platforms in rapid cycling.

    Science.gov (United States)

    Eischeid, Anne C; Kasko, Sasha M

    2015-01-01

    Real-time PCR has been used widely in numerous fields. In food safety, it has been applied to detection of microbes and other contaminants, including food allergens. Interest in rapid (fast) cycling real-time PCR has grown because it yields results in less time than does conventional cycling. However, fast cycling can adversely affect assay performance. Here we report on tests of commercial master mixes specifically designed for fast real-time PCR using a shrimp allergen assay we previously developed and validated. The objective of this work was to determine whether specialized commercial master mixes lead to improved assay performance in rapid cycling. Real-time PCR assays were carried out using four different master mixes and two different rapid cycling protocols. Results indicated that specialized master mixes did yield quality results. In many cases, linear ranges spanned up to 7 orders of magnitude, R(2) values were at least 0.95, and reaction efficiencies were within or near the optimal range of 90 to 110%. In the faster of the two rapid cycling protocols tested, assay performance and PCR amplification were markedly better for the shorter PCR product. In conclusion, specialized commercial master mixes were effective as part of rapid cycling protocols, but conventional cycling as used in our previous work is more reliable for the shrimp assay tested.

  6. Comparison of benthos and plankton for selected areas of concern and non-areas of concern in western Lake Michigan Rivers and Harbors in 2012

    Science.gov (United States)

    Eikenberry, Barbara C. Scudder; Bell, Amanda H.; Templar, Hayley A.; Burns, Daniel J.

    2016-07-25

    Recent data are lacking to assess whether impairments still exist at four of Wisconsin’s largest Lake Michigan harbors that were designated as Areas of Concern (AOCs) in the late 1980s due to sediment contamination and multiple Beneficial Use Impairments (BUIs), such as those affecting benthos (macroinvertebrates) and plankton (zooplankton and phytoplankton) communities. During three seasonal sampling events (“seasons”) in May through August 2012, the U.S. Geological Survey collected sediment benthos and water plankton at the four AOCs as well as six less-degraded non-AOCs along the western Lake Michigan shoreline to assess whether AOC communities were degraded in comparison to non-AOC communities. The four AOCs are the Lower Menominee River, the Lower Green Bay and Fox River, the Sheboygan River, and the Milwaukee Estuary. Due to their size and complexity, multiple locations or “subsites” were sampled within the Lower Green Bay and Fox River AOC (Lower Green Bay, the Fox River near Allouez, and the Fox River near De Pere) and within the Milwaukee Estuary AOC (the Milwaukee River, the Menomonee River, and the Milwaukee Harbor) and single locations were sampled at the other AOCs and non-AOCs. The six non-AOCs are the Escanaba River in Michigan, and the Oconto River, Ahnapee River, Kewaunee River, Manitowoc River, and Root River in Wisconsin. Benthos samples were collected by using Hester-Dendy artificial substrates deployed for 30 days and by using a dredge sampler; zooplankton were collected by net and phytoplankton by whole-water sampler. Except for the Lower Green Bay and Milwaukee Harbor locations, communities at each AOC were compared to all non-AOCs as a group and to paired non-AOCs using taxa relative abundances and metrics, including richness, diversity, and an Index of Biotic Integrity (IBI, for Hester-Dendy samples only). Benthos samples collected during one or more seasons were rated as degraded for at least one metric at all AOCs. In the

  7. Comparison of benthos and plankton for selected areas of concern and non-areas of concern in western Lake Michigan Rivers and Harbors in 2012

    Science.gov (United States)

    Eikenberry, Barbara C. Scudder; Bell, Amanda H.; Templar, Hayley A.; Burns, Daniel J.

    2016-07-25

    Recent data are lacking to assess whether impairments still exist at four of Wisconsin’s largest Lake Michigan harbors that were designated as Areas of Concern (AOCs) in the late 1980s due to sediment contamination and multiple Beneficial Use Impairments (BUIs), such as those affecting benthos (macroinvertebrates) and plankton (zooplankton and phytoplankton) communities. During three seasonal sampling events (“seasons”) in May through August 2012, the U.S. Geological Survey collected sediment benthos and water plankton at the four AOCs as well as six less-degraded non-AOCs along the western Lake Michigan shoreline to assess whether AOC communities were degraded in comparison to non-AOC communities. The four AOCs are the Lower Menominee River, the Lower Green Bay and Fox River, the Sheboygan River, and the Milwaukee Estuary. Due to their size and complexity, multiple locations or “subsites” were sampled within the Lower Green Bay and Fox River AOC (Lower Green Bay, the Fox River near Allouez, and the Fox River near De Pere) and within the Milwaukee Estuary AOC (the Milwaukee River, the Menomonee River, and the Milwaukee Harbor) and single locations were sampled at the other AOCs and non-AOCs. The six non-AOCs are the Escanaba River in Michigan, and the Oconto River, Ahnapee River, Kewaunee River, Manitowoc River, and Root River in Wisconsin. Benthos samples were collected by using Hester-Dendy artificial substrates deployed for 30 days and by using a dredge sampler; zooplankton were collected by net and phytoplankton by whole-water sampler. Except for the Lower Green Bay and Milwaukee Harbor locations, communities at each AOC were compared to all non-AOCs as a group and to paired non-AOCs using taxa relative abundances and metrics, including richness, diversity, and an Index of Biotic Integrity (IBI, for Hester-Dendy samples only). Benthos samples collected during one or more seasons were rated as degraded for at least one metric at all AOCs. In the

  8. Detection of knockdown resistance (kdr mutations in Anopheles gambiae: a comparison of two new high-throughput assays with existing methods

    Directory of Open Access Journals (Sweden)

    Ball Amanda

    2007-08-01

    Full Text Available Abstract Background Knockdown resistance (kdr is a well-characterized mechanism of resistance to pyrethroid insecticides in many insect species and is caused by point mutations of the pyrethroid target site the para-type sodium channel. The presence of kdr mutations in Anopheles gambiae, the most important malaria vector in Africa, has been monitored using a variety of molecular techniques. However, there are few reports comparing the performance of these different assays. In this study, two new high-throughput assays were developed and compared with four established techniques. Methods Fluorescence-based assays based on 1 TaqMan probes and 2 high resolution melt (HRM analysis were developed to detect kdr alleles in An. gambiae. Four previously reported techniques for kdr detection, Allele Specific Polymerase Chain Reaction (AS-PCR, Heated Oligonucleotide Ligation Assay (HOLA, Sequence Specific Oligonucleotide Probe – Enzyme-Linked ImmunoSorbent Assay (SSOP-ELISA and PCR-Dot Blot were also optimized. The sensitivity and specificity of all six assays was then compared in a blind genotyping trial of 96 single insect samples that included a variety of kdr genotypes and African Anopheline species. The relative merits of each assay was assessed based on the performance in the genotyping trial, the length/difficulty of each protocol, cost (both capital outlay and consumable cost, and safety (requirement for hazardous chemicals. Results The real-time TaqMan assay was both the most sensitive (with the lowest number of failed reactions and the most specific (with the lowest number of incorrect scores. Adapting the TaqMan assay to use a PCR machine and endpoint measurement with a fluorimeter showed a slight reduction in sensitivity and specificity. HRM initially gave promising results but was more sensitive to both DNA quality and quantity and consequently showed a higher rate of failure and incorrect scores. The sensitivity and specificity of AS

  9. Total vitamin D assay comparison of the Roche Diagnostics "Vitamin D total" electrochemiluminescence protein binding assay with the Chromsystems HPLC method in a population with both D2 and D3 forms of vitamin D.

    Science.gov (United States)

    Abdel-Wareth, Laila; Haq, Afrozul; Turner, Andrew; Khan, Shoukat; Salem, Arwa; Mustafa, Faten; Hussein, Nafiz; Pallinalakam, Fasila; Grundy, Louisa; Patras, Gemma; Rajah, Jaishen

    2013-03-22

    This study compared two methods of assaying the 25-hydroxylated metabolites of cholecalciferol (vitamin D3) and ergocalciferol (vitamin D2). A fully automated electrochemiluminescence assay from Roche Diagnostics and an HPLC based method from Chromsystems were used to measure vitamin D levels in surplus sera from 96 individuals, where the majority has the D2 form of the vitamin. Deming regression, concordance rate, correlation and Altman Bland agreement were performed. Seventy two subjects (75%) had a D2 concentration >10 nmol/L while the remaining twenty four subjects had vitamin D2 concentration of less than 10 nmol/L by HPLC. Overall, the Roche Diagnostics method showed a negative bias of -2.59 ± 4.11 nmol/L on the e602 as compared to the HPLC with a concordance rate of 84%. The concordance rate was 91% in samples with D2 of less than 10 nmol/L and 82% in those with D2 concentration >10 nmol/L. The overall correlation had an r value of 0.77. The r value was higher in samples with D2 levels of less than 10 nmol/L, r = 0.96, as compared to those with D2 values of greater than 10 nmol/L, r = 0.74. The observed bias had little impact on clinical decision and therefore is clinically acceptable.

  10. Total Vitamin D Assay Comparison of the Roche Diagnostics “Vitamin D Total” Electrochemiluminescence Protein Binding Assay with the Chromsystems HPLC Method in a Population with both D2 and D3 forms of Vitamin D

    Directory of Open Access Journals (Sweden)

    Gemma Patras

    2013-03-01

    Full Text Available This study compared two methods of assaying the 25-hydroxylated metabolites of cholecalciferol (vitamin D3 and ergocalciferol (vitamin D2. A fully automated electrochemiluminescence assay from Roche Diagnostics and an HPLC based method from Chromsystems were used to measure vitamin D levels in surplus sera from 96 individuals, where the majority has the D2 form of the vitamin. Deming regression, concordance rate, correlation and Altman Bland agreement were performed. Seventy two subjects (75% had a D2 concentration >10 nmol/L while the remaining twenty four subjects had vitamin D2 concentration of less than 10 nmol/L by HPLC. Overall, the Roche Diagnostics method showed a negative bias of −2.59 ± 4.11 nmol/L on the e602 as compared to the HPLC with a concordance rate of 84%. The concordance rate was 91% in samples with D2 of less than 10 nmol/L and 82% in those with D2 concentration >10 nmol/L. The overall correlation had an r value of 0.77. The r value was higher in samples with D2 levels of less than 10 nmol/L, r = 0.96, as compared to those with D2 values of greater than 10 nmol/L, r = 0.74. The observed bias had little impact on clinical decision and therefore is clinically acceptable.

  11. Rapid drug-susceptibility testing of Mycobacterium tuberculosis clinical isolates to first-line antitubercular drugs by nitrate reductase assay: A comparison with proportion method.

    Science.gov (United States)

    Kohli, Amrish; Bashir, Gulnaz; Fatima, Akeela; Jan, Abiroo; Wani, Nayeem-U-Din; Ahmad, Junaid

    2016-12-01

    Early initiation of therapy in patients with tuberculosis is imperative for its control. Conventional methods of susceptibility testing such as the proportion method (PM) require visual detection and counting of colonies that takes up to 6weeks. Rapid and simple phenotypic methods that have been endorsed by the World Health Organization can serve as alternatives. In this study, we evaluated the colorimetric nitrate reductase assay, which utilizes the detection of nitrate reduction as an indicator of growth much earlier compared with PM (within 7-14days). The susceptibility of 75 clinical isolates of Mycobacterium tuberculosis to four first-line antitubercular drugs was tested by nitrate reductase assay and compared with the standard PM. In this assay, inoculation was done on both drug-free and drug-containing Löwenstein-Jensen medium containing sodium nitrate. After incubation for 7-14days, reduction to nitrite was taken as an indicator of growth, which was detected by color change on addition of Griess reagent. Agreement between nitrate reductase assay and PM was 100% for rifampicin, 97.30% for isoniazid, 93.30% for streptomycin, and 98.60% for ethambutol. Cost/isolate with this assay was found to be approximately two times lesser than that of PM. All results were obtained in 7-14days by nitrate reductase assay, which was significantly rapid compared with 42days taken for obtaining results by PM. Nitrate reductase assay can be used as a rapid and inexpensive method for drug-susceptibility testing of M. tuberculosis for first-line antitubercular drugs without compromising accuracy of standard methods. Copyright © 2016 Asian-African Society for Mycobacteriology. Published by Elsevier Ltd. All rights reserved.

  12. Comparison of an array of in vitro assays for the assessment of the estrogenic potential of natural and synthetic estrogens, phytoestrogens and xenoestrogens.

    Science.gov (United States)

    Gutendorf, B; Westendorf, J

    2001-09-14

    Many chemicals in surface waters and sediments have recently been discovered to have estrogenic/antiestrogenic activity. Among these compounds, known as 'endocrine disrupters', are natural and synthetic hormones, phytoestrogenes and a variety of industrial chemicals, such as certain detergents and pesticides. These substances are supposed to affect the development and reproduction in wildlife and humans and may also be involved in the induction of cancer. In order to assess the estrogenic/antiestrogenic potential of pure compounds and complex environmental samples we compared an array of in vitro test systems, (i) two luciferase reporter gene assays using transgenic human MVLN cells (derived from MCF-7 cells) and HGELN cells (derived from HeLa cells); (ii) a competitive binding assay with recombinant human estrogen receptors (ER) alpha and beta; and (iii) a proliferation assay with MCF7-cells (E-Screen). The sensitivity of the assays for 17-beta-estradiol decreased in the order: MVLN-cells=E-Screen>HGELN-cells>binding to ER-alpha>binding to ER-beta. A good correlation was obtained between the estrogenic potencies of 11 compounds (17-beta-estradiol (E(2)), estrone (E(1)), estriol (E(3)), ethinylestradiol (EE(2)), diethylstilbestrol (DES), coumestrol, beta-sitosterol, genistein, 4-nonylphenol, 4-octylphenol, bisphenol A) in the three tissue culture assays. The relative potencies of the compounds obtained by the cell free binding assays were one to two orders of magnitude higher compared with the cell culture assays. The phytoestrogens showed a preference to bind to ER-beta, but only genistein showed a much lower activity in the E-Screen (growth induction in breast cancer cells) compared with the luciferase induction in MVLN and HGELN-cells.

  13. A comparison of titers of anti-Brucella antibodies of naturally infected and healthy vaccinated cattle by standard tube agglutination test, microtiter plate agglutination test, indirect hemagglutination assay, and indirect enzyme-linked immunosorbent assay

    Directory of Open Access Journals (Sweden)

    Anju Mohan

    2016-07-01

    Full Text Available Aim: We determined the antibody response in cattle naturally infected with brucellosis and normal healthy adult cattle vaccinated during calf hood with strain 19. Materials and Methods: The antibody titers were measured by standard tube agglutination test (STAT, microtiter plate agglutination test (MAT, indirect hemagglutination assay (IHA, and indirect enzyme-linked immunosorbent assay (iELISA as per standard protocols. Results: The mean STAT titers were 1.963±0.345 in infected cattle and 1.200±0.155 in healthy vaccinated cattle. The difference was extremely significant (p<0.0001. The mean MAT titers were 2.244±0.727 in infected cattle and 1.200±0.155 in healthy vaccinated cattle. The difference was very significant (p<0.005. The mean IHA titers in infected cattle were 2.284±0.574, and those in healthy vaccinated cattle were 1.200±0.155. The difference was extremely significant (p=0.0002. However, the difference in mean iELISA titers of infected cattle (1.3678±0.014 and healthy vaccinated cattle (1.367±0.014 was non-significant. The infected animals showed very high titers of agglutinating antibodies compared to the vaccinated animals. However, it cannot be ascertained whether these antibodies are due to vaccine or response to infection. Since the infected animals had been vaccinated earlier, the current infection may suggest that vaccination was unable to induce protective levels of antibody. The heightened antibody response after infection may also indicate a secondary immune response to the antigens common to the vaccine strain and wild Brucella organisms. Conclusion: The brucellosis infected animals showed very high titers of agglutinating antibodies compared to the vaccinated animals.

  14. Quantitation of HTLV-I proviral load by a real-time PCR assay using SYBR Green: comparison of two methods for DNA isolation.

    Science.gov (United States)

    Altamirano, Natalia Andrea; Rocco, Carlos; Aulicino, Paula; Sen, Luisa; Mangano, Andrea

    2010-12-01

    A real-time quantitative PCR (qPCR) assay using SYBR Green was developed to determine HTLV-I proviral load (pVL) in peripheral blood mononuclear cells (PBMCs), and its performance was evaluated with samples processed as cell lysates and DNA isolated by salting out. Primers targeting the pol region were standardized against the MT2 cell line and HTLV-I copy number was normalized to the amount of cellular DNA by quantitation of the albumin gene. The sensitivity, specificity and reproducibility of the qPCR were assessed in the two methods used for DNA processing. The assay had a limit of detection of 400 HTLV-I copies/10(6) PBMCs for both methods, with a broad range of quantitation (2.6log(10) to >5log(10)), and without cross-reactivity with HTLV-II or with HIV-1. The inter- and intra-assay coefficients of variation were less than 2.4%. HTLV-I pVL quantitation in seven blood donor samples processed as either cell lysates or isolated DNA by salting out showed a strong linear correlation and no difference in the calculated pVL (Fisher's exact test, p>0.05). The assay was found to be a low cost, robust and reproducible assay for quantifying HTLV-I pVL in samples processed as cell lysates or as isolated DNA.

  15. Limited ability of circulating anti-Müllerian hormone to predict dominant follicular recruitment in PCOS women treated with clomiphene citrate: a comparison of two different assays.

    Science.gov (United States)

    Vaiarelli, Alberto; Drakopoulos, Panagiotis; Blockeel, Christophe; De Vos, Michel; van de Vijver, Arne; Camus, Michel; Cosyns, Stefan; Tournaye, Herman; Polyzos, Nikolaos P

    2016-01-01

    The present retrospective cohort study was conducted to investigate whether serum anti-Müllerian hormone (AMH) levels, determined by either the Immunotech (IOT) or the second generation (Gen II) assay, can predict follicular recruitment in women with polycystic ovary syndrome (PCOS) undergoing ovulation induction with clomiphene citrate (CC). Patients received 50 mg CC daily for ovulation induction followed by natural intercourse or intrauterine insemination. Overall, 84 women had their serum AMH levels tested before treatment [42 patients with Immunotech (IOT), and 42 patients with the Gen II assay]. The primary outcome was to determine dominant follicle (>10 mm) recruitment in relation to AMH levels. Thirty-three (79%) patients in the IOT and 34 (81%) patients in the Gen II assay group developed a dominant follicle within 15 days after initiation of CC. Circulating AMH levels did not differ between women with or without dominant follicular recruitment in the both groups. By using either the AMH IOT or the Gen II assay, serum AMH levels were not predictive of the development of a dominant follicle. In conclusion, serum AMH levels measured by IOT or Gen II assay, has limited value to predict PCOS patients who will develop a dominant follicle following ovulation induction with CC.

  16. Detection limits and cost comparisons of human- and gull-associated conventional and quantitative PCR assays in artificial and environmental waters

    Science.gov (United States)

    Riedel, Timothy E.; Zimmer-Faust, Amity G.; Thulsiraj, Vanessa; Madi, Tania; Hanley, Kaitlyn T.; Ebentier, Darcy L.; Byappanahalli, Muruleedhara N.; Layton, Blythe; Raith, Meredith; Boehm, Alexandria B.; Griffith, John F.; Holden, Patricia A.; Shanks, Orin C.; Weisberg, Stephen B.; Jay, Jennifer A.

    2014-01-01

    Some molecular methods for tracking fecal pollution in environmental waters have both PCR and quantitative PCR (qPCR) assays available for use. To assist managers in deciding whether to implement newer qPCR techniques in routine monitoring programs, we compared detection limits (LODs) and costs of PCR and qPCR assays with identical targets that are relevant to beach water quality assessment. For human-associated assays targeting Bacteroidales HF183 genetic marker, qPCR LODs were 70 times lower and there was no effect of target matrix (artificial freshwater, environmental creek water, and environmental marine water) on PCR or qPCR LODs. The PCR startup and annual costs were the lowest, while the per reaction cost was 62% lower than the Taqman based qPCR and 180% higher than the SYBR based qPCR. For gull-associated assays, there was no significant difference between PCR and qPCR LODs, target matrix did not effect PCR or qPCR LODs, and PCR startup, annual, and per reaction costs were lower. Upgrading to qPCR involves greater startup and annual costs, but this increase may be justified in the case of the human-associated assays with lower detection limits and reduced cost per sample.

  17. EV71病毒核酸快速检测方法的比较%Comparison of Different Molecular Assays for the Rapid Detection of Enterovirus 71(EV71)

    Institute of Scientific and Technical Information of China (English)

    卫海燕; 黄学勇; 许玉玲; 马宏; 陈豪敏; 许汴利

    2012-01-01

    Molecular detection of enterovirus (EV)71 RNA based on PCR methods is a quick and sensitive approach. At present, different PCR-based methods for EV71 RNA detection are available, but comparisons of results obtained using different approaches are limited. This study is to compare the analytical sensitivity and specificity of different real-time reverse transcription-polymerase chain reaction (rRT-PCR) and conventional reverse transcription-polymerase chain reaction (cRT-PCR) assays for enterovirus and EV71 detection, Altogether, three rRT-PCR assays and one cRT-PCR assay targeting the 5'UTR gene for universal detection of enterovirus; two rRT-PCR assays andone cRT-PCR assay targeting the VP1 gene for specific detection of EV 71 were examined. All assays showed good specificity. The detection sensitivity ranged from 8. 19 ×10 to 8. 19 ×105 copy equivalents. In general, rRT-PCR assays were more sensitive than cRT-PCR assays. All rRT-PCR assays showed 100% sensitivity for clinical specimens.%PCR方法检测肠道病毒71型(EV71)的分子诊断技术十分快速敏感.目前已有多篇文献报道了不同的检测EV71的PCR方法,但对不同方法检测结果的比较还少有报道.本研究将检测肠道病毒通用5′UTR基因的3种定量PCR(rRT-PCR)方法和1种普通PCR(cRT-PCR)方法和特异性检测EV71的2种rRT-PCR和1种cRT-PCR方法进行特异性和敏感性比较评估.所有试验方法都有较好的特异性,无交叉反应.核酸最低检测限值为从8.19× 101~8.19×105个拷贝,rRT-PCR比cRT-PCR更敏感.所有的rRT-PCR方法检测50份临床标本时都较敏感.

  18. Angiogenesis Assays.

    Science.gov (United States)

    Nambiar, Dhanya K; Kujur, Praveen K; Singh, Rana P

    2016-01-01

    Neoangiogenesis constitutes one of the first steps of tumor progression beyond a critical size of tumor growth, which supplies a dormant mass of cancerous cells with the required nutrient supply and gaseous exchange through blood vessels essentially needed for their sustained and aggressive growth. In order to understand any biological process, it becomes imperative that we use models, which could mimic the actual biological system as closely as possible. Hence, finding the most appropriate model is always a vital part of any experimental design. Angiogenesis research has also been much affected due to lack of simple, reliable, and relevant models which could be easily quantitated. The angiogenesis models have been used extensively for studying the action of various molecules for agonist or antagonistic behaviour and associated mechanisms. Here, we have described two protocols or models which have been popularly utilized for studying angiogenic parameters. Rat aortic ring assay tends to bridge the gap between in vitro and in vivo models. The chorioallantoic membrane (CAM) assay is one of the most utilized in vivo model system for angiogenesis-related studies. The CAM is highly vascularized tissue of the avian embryo and serves as a good model to study the effects of various test compounds on neoangiogenesis.

  19. Determination of anti-endomysium IgA antibodies in the diagnosis of celiac disease: Comparison of a novel ELISA-based assay with conventional immunofluorescence

    Institute of Scientific and Technical Information of China (English)

    Dennis CW Poland; Huib Ceelie; Rob B Dinkelaar; Cornelis Beijer

    2006-01-01

    AIM: To evaluate the novel anti-endomysium (anti-EMA)detection based on ELISA.METHODS: Anti-EMA IgA was measured by a novel ELISA in 196 patients with gastrointestinal symptoms and suspected mal-absorption. Data were compared with those obtained by the conventional IF test.RESULTS: A good concordance of 98% was found between these two assays. In sera of 161 patients (82%)both assays tested negative whereas in sera of 31 patients (16%) both assays tested positive for the presence of anti-EMA antibodies. Discrepancies between EMAELISA and EMA-immunofluorescence (IF) were found in only 4 patients (2%).CONCLUSION: This ELISA can replace IF for the detection of anti-EMA antibodies and provide clinicians with an excellent tool to screen for celiac disease in patients with gastrointestinal complaints.

  20. In vitro micronucleus assay for the analysis of total particulate matter in cigarette smoke: comparison of flow cytometry and laser scanning cytometry with microscopy.

    Science.gov (United States)

    Yao, Jianhua; Gao, Qian; Mi, Qili; Li, Xuemei; Miao, Mingming; Cheng, Peng; Luo, Ying

    2013-08-15

    The possible genotoxicity of the total particulate matter (TPM) in cigarette smoke has typically been evaluated using the in vitro micronucleus assay. In recent years, automated scoring techniques have been developed to replace the manual counting process in this assay. However, these automated scoring techniques have not been applied in routine genotoxicity assays for the analysis of TPM to improve the assay efficiency. Chinese hamster ovary (CHO) cells were treated with TPM produced from 14 types of cigarettes at five concentrations (25-200μg/ml) without exogenous metabolic activation. The three following methods were used to score the micronucleus (MN) frequency: (a) flow cytometry with SYTOX and EMA dyes, which differentially stain micronuclei and apoptotic/necrotic chromatin to enhance assay reliability; (b) laser scanning cytometry with FITC and PI dyes, which is a system that combines the analytical capabilities of flow and image cytometry; and (c) visual microcopy with Giemsa dye. The test results obtained using the three methods were compared using correlation analysis. The key findings for this set of compounds include the following: (a) both flow cytometry- and laser scanning cytometry-based methods were effective for MN identification, (b) the three scoring methods could detect dose-dependent micronucleus formation for the 14 types of TPM, and (c) the MN frequencies that were measured in the same samples by flow cytometry, laser scanning cytometry, and visual microscopy were highly correlated, and there were no significant differences (p>0.05). In conclusion, both flow cytometry and laser scanning cytometry can be used to evaluate the MN frequency induced by TPM without exogenous metabolic activation. The simpler and faster processing and the high correlation of the results make these two automatic methods appropriate tools for use in in vitro micronucleus assays for the analysis of TPM using CHO cells.

  1. HIV incidence in rural South Africa: comparison of estimates from longitudinal surveillance and cross-sectional cBED assay testing.

    Directory of Open Access Journals (Sweden)

    Till Bärnighausen

    Full Text Available BACKGROUND: The BED IgG-Capture Enzyme Immunoassay (cBED assay, a test of recent HIV infection, has been used to estimate HIV incidence in cross-sectional HIV surveys. However, there has been concern that the assay overestimates HIV incidence to an unknown extent because it falsely classifies some individuals with non-recent HIV infections as recently infected. We used data from a longitudinal HIV surveillance in rural South Africa to measure the fraction of people with non-recent HIV infection who are falsely classified as recently HIV-infected by the cBED assay (the long-term false-positive ratio (FPR and compared cBED assay-based HIV incidence estimates to longitudinally measured HIV incidence. METHODOLOGY/PRINCIPAL FINDINGS: We measured the long-term FPR in individuals with two positive HIV tests (in the HIV surveillance, 2003-2006 more than 306 days apart (sample size n = 1,065. We implemented four different formulae to calculate HIV incidence using cBED assay testing (n = 11,755 and obtained confidence intervals (CIs by directly calculating the central 95(th percentile of incidence values. We observed 4,869 individuals over 7,685 person-years for longitudinal HIV incidence estimation. The long-term FPR was 0.0169 (95% CI 0.0100-0.0266. Using this FPR, the cross-sectional cBED-based HIV incidence estimates (per 100 people per year varied between 3.03 (95% CI 2.44-3.63 and 3.19 (95% CI 2.57-3.82, depending on the incidence formula. Using a long-term FPR of 0.0560 based on previous studies, HIV incidence estimates varied between 0.65 (95% CI 0.00-1.32 and 0.71 (95% CI 0.00-1.43. The longitudinally measured HIV incidence was 3.09 per 100 people per year (95% CI 2.69-3.52, after adjustment to the sex-age distribution of the sample used in cBED assay-based estimation. CONCLUSIONS/SIGNIFICANCE: In a rural community in South Africa with high HIV prevalence, the long-term FPR of the cBED assay is substantially lower than previous estimates. The c

  2. MICROBIOLOGICAL ASSAY FOR VITAMIN B

    OpenAIRE

    Bishnoi Kapil*, , ,; Kataria Mahesh; Singhal Vipin; Gupta Deepika

    2012-01-01

    Micronutrients added to foods are analyzed using various procedures depending on their nature and properties. The microbiological assays are better than chemical method because any suitable change in vitamin molecule which may not be detected by chemical method will be revealed by change in microbial activity. The microbiological assay of vitamins is based upon the comparison of the stimulation of growth of bacteria by measured concentration of vitamin with that produced by known concentratio...

  3. Comparison of PCR-ELISA and LightCycler real-time PCR assays for detecting Salmonella spp. in milk and meat samples

    DEFF Research Database (Denmark)

    Perelle, Sylvie; Dilasser, Françoise; Malorny, Burkhard

    2004-01-01

    In a previous study, we reported the performance of a PCR assay amplifying 285-bp of the invA gene of Salmonella spp. through an international ring-trial involving four participating laboratories [Int. J. Food Microbiol. 89 (2003) 241]. Based on the validated set of primers and recent advancement...

  4. Impact of Assay conditions on activity estimate and kinetics comparison of Aspergillus niger PhyA and Escherichia coli AppA2 phytases

    Science.gov (United States)

    This study was to compare three phytase activity assays and kinetics of Aspergillus niger PhyA and Escherichia coli AppA2 phytases expressed in Pichia pastoris at the observed stomach pH of 3.5. In Experiment 1, equivalent phytase activities in the crude preparations of PhyA and AppA2 were tested ...

  5. Comparison of three different in vitro mutation assays used for the investigation of cytochrome P450-mediated mutagenicity of nitro-polycyclic aromatic hydrocarbons

    NARCIS (Netherlands)

    Kappers, W.A.; Och, F.M.M. van; Groene, E.M. de; Horbach, G.J.

    2000-01-01

    Three different in vitro mutation assays were used to investigate the involvement of cytochrome P450 enzymes in the activation of the nitro- polycyclic aromatic hydrocarbons (nitroPAHs) 1-nitropyrene and 2- nitrofluorene and their reduced metabolites amino-polycyclic aromatic hydrocarbons (aminoPAHs

  6. Analytical and clinical comparison of Elecsys syphilis (Roche(®)) - Architect syphilis TP and reformulated Architect syphilis TP (Abbott(®)) assay.

    Science.gov (United States)

    De Keukeleire, Steven; Desmet, Stefanie; Lagrou, Katrien; Oosterlynck, Julie; Verhulst, Manon; Van Besien, Jessica; Saegeman, Veroniek; Reynders, Marijke

    2017-03-01

    The performance of Elecsys Syphilis was compared to Architect Syphilis TP and Reformulated Architect Syphilis TP. The overall sensitivity and specificity were 98.4% and 99.5%, 97.7% and 97.1%, and 99.2% and 99.7% respectively. The assays are comparable and considered adequate for syphilis screening.

  7. Characteristic comparison of triglyceride-rich remnant lipoprotein measurement between a new homogenous assay (RemL-C and a conventional immunoseparation method (RLP-C

    Directory of Open Access Journals (Sweden)

    Saikawa Shinichi

    2008-05-01

    Full Text Available Abstract Background Increased serum remnant lipoproteins are supposed to predict cardiovascular disease in addition to increased LDL. A new homogenous assay for remnant lipoprotein-cholesterol (RemL-C has been developed as an alternative to remnant-like particle-cholesterol (RLP-C, an immunoseparation assay, widely used for the measurement of remnant lipoprotein cholesterol. Methods We evaluated the correlations and data validation between the 2 assays in 83 subjects (49 men and 34 women without diabetes, hypertension and medications for hyperlipidemia, diabetes, and hypertension, and investigated the characteristics of remnant lipoproteins obtained by the two methods (RLP-C and RemL-C and their relationships with IDL-cholesterol determined by our developed HPLC method. Results A positive correlation was significantly found between the two methods (r = 0.853, 95%CI 0.781–0.903, p RLP-C level. RemL-C (r = 0.339, 95%CI 0.152–0.903; p = 0.0005 significantly correlated with IDL-cholesterol, but not RLP-C (r = 0.17, 95%CI -0.047–0.372; p = 0.1237 in all the samples (n = 83. Conclusion These results suggest that there is generally a significant correlation between RemL-C and RLP-C. However, RemL-C assay is likely to reflect IDL more closely than RLP-C.

  8. Comparison of the inhibition of urokinase-type plasminogen activator (u-PA) activity by monoclonal antibodies specific for u-PA as assessed by different assays

    NARCIS (Netherlands)

    Boheemen, P.A. van; Hoogen, N.M. van den; Koolwijk, N.

    1995-01-01

    Six murine monoclonal antibodies (MAbs) specific for urokinase-type plasminogen activator (u-PA) were tested for their ability to inhibit u-PA activity in three different assays with respect to amidolytic activity, plasminogen activation and fibrinolytic activity. Two of the MAbs were able to inhibi

  9. Tuberculosis screening in patients with psoriasis before antitumour necrosis factor therapy : comparison of an interferon-gamma release assay vs. tuberculin skin test

    NARCIS (Netherlands)

    Laffitte, E.; Janssens, J. P.; Roux-Lombard, P.; Thielen, A. M.; Barde, C.; Marazza, G.; Panizzon, R. G.; Saurat, J-H.

    2009-01-01

    Background Antitumour necrosis factor (anti-TNF) treatments may reactivate latent tuberculosis infection (LTBI). For detecting LTBI, the tuberculin skin test (TST) has low sensitivity and specificity. Interferon-gamma release assays (IGRA) have been shown to be more sensitive and specific than TST.

  10. Wide dynamic range of surface-plasmon-resonance-based assay for Hepatitis-B-surface-antigen-antibody optimal detection in comparison with ELISA.

    Science.gov (United States)

    Tam, Yew Joon; Zeenathul, Nazariah Allaudin; Rezaei, Morvarid Akhavan; Mustafa, Nor Hidayah; Azmi, Mohd Lila Mohd; Bahaman, Abdul Rani; Lo, Sewn Cen; Tan, Joo Shun; Hani, Homayoun; Rasedee, Abdullah

    2016-08-10

    Limit of detection (LOD), limit of quantification (LOQ) and the dynamic range of detection of hepatitis B surface antigen antibody (anti-HBs) using surface plasmon resonance (SPR) chip-based approach with Pichia pastoris derived recombinant hepatitis B surface antigen (HBsAg) as recognition element were established through the scouting for optimal conditions for the improvement of immobilization efficiency and in the use of optimal regeneration buffer. Recombinant HBsAg was immobilized onto the sensor surface of CM5 chip at a concentration of 150 mg/L, in sodium acetate buffer at pH 4 with added 0.6% Triton X-100. Regeneration solution of 20 mM HCl was optimally found to effectively unbind analytes from the ligand, thus allowing for multiple screening cycles. A dynamic range of detection of ∼0.00098 to 0.25 mg/L was obtained and a 7-fold higher LOD, as well as a 2-fold increase in coefficient of variance (CV) of the replicated results, were shown as compared to enzyme-linked immunosorbent assay (ELISA). Evaluation of the assay for specificity showed no cross-reactivity with other antibodies tested. The ability of SPR chip-based assay and ELISA to detect anti-HBs in human serum was comparable, indicating that the SPR chip-based assay with its multiple screening capacity has greater advantage over ELISA. This article is protected by copyright. All rights reserved.

  11. Comparison of commercial and in-house Real-time PCR assays for quantification of Epstein-Barr virus (EBV DNA in plasma

    Directory of Open Access Journals (Sweden)

    Pollara Caterina

    2007-03-01

    Full Text Available Abstract Background Epstein-Barr virus (EBV DNA load monitoring is known to be useful for the diagnosis and monitoring of EBV-associated diseases. The aim of this study is to compare the performance of two real-time PCR assays for EBV DNA: a commercial kit as the Q-EBV Real-Time System (Q-EBV PCR, Amplimedical, Turin, Italy and an in-house assay (EBV RQ-PCR. Results The range of linearity and the degree of precision of the two assays were similar. The clinical sensitivity of Q-EBV PCR was higher for reference samples containing less than 1,000 EBV DNA copies/ml. The absolute quantitative results of the two methods were statistically correlated (R2 = 0.7789; p Conclusion Both the commercial and the in-house assay may be appropriate for clinical use, but common standards are advisable for comparable absolute values, as these would improve the clinical utility of EBV DNA load measurement.

  12. Comparison of illumigene Group A Streptococcus Assay with Culture of Throat Swabs from Children with Sore Throats in the New Zealand School-Based Rheumatic Fever Prevention Program.

    Science.gov (United States)

    Upton, Arlo; Bissessor, Liselle; Farrell, Elizabeth; Shulman, Stanford T; Zheng, Xiaotian; Lennon, Diana

    2016-01-01

    Group A streptococcal (GAS) pharyngitis is a particularly important condition in areas of New Zealand where the incidence of acute rheumatic fever remains unacceptably high. Prompt diagnosis and treatment of GAS pharyngitis are cornerstones of the Rheumatic Fever Prevention Programme, but these are hindered by the turnaround time of culture. Tests with excellent performance and rapid turnaround times are needed. For this study, throat swabs (Copan ESwabs) were collected from schoolchildren self-identifying with a sore throat. Samples were tested by routine culture and the illumigene GAS assay using loop-mediated isothermal amplification. Discrepant results were resolved by retesting of the same specimen by an alternative molecular assay. Seven hundred fifty-seven throat swab specimens were tested by both methods. The performance characteristics of the illumigene assay using culture on blood agar as the "gold standard" and following discrepancy analysis were as follows: sensitivity, 82% and 87%, respectively; specificity, 93% and 98%, respectively; positive predictive value, 61% and 88%, respectively; and negative predictive value, 97% and 97%, respectively. In our unique setting of a school-based throat swabbing program, the illumigene assay did not perform quite as well as described in previous reports. Despite this, its improved sensitivity and rapid turnaround time compared with those of culture are appealing.

  13. Iriflophenone-3-C-glucoside from Cyclopia genistoides: isolation and quantitative comparison of antioxidant capacity with mangiferin and isomangiferin using on-line HPLC antioxidant assays.

    Science.gov (United States)

    Malherbe, Christiaan J; Willenburg, Elize; de Beer, Dalene; Bonnet, Susan L; van der Westhuizen, Jan H; Joubert, Elizabeth

    2014-03-01

    The benzophenone, iriflophenone-3-C-glucoside, was isolated from Cyclopia genistoides using a combination of fluid-fluid extraction, high performance counter-current chromatography (HPCCC) and semi-preparative high performance liquid chromatography (HPLC). The microplate oxygen radical absorbance capacity (ORAC) assay, with fluorescein as probe, was adapted for use in an on-line HPLC configuration. The method was validated using a mixture of authentic standards including iriflophenone-3-C-glucoside, and the xanthones, mangiferin and isomangiferin. Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid) was included in the mixture for calculation of Trolox equivalent antioxidant capacity (TEAC) values. Using the on-line HPLC-ORAC assay, as well as 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS(+)) on-line assays, the antioxidant activity of iriflophenone-3-C-glucoside and isomangiferin was demonstrated for the first time. Iriflophenone-3-C-glucoside presented no radical scavenging ability against DPPH, but scavenged ABTS(+) and peroxyl radicals (TEACABTS of 1.04 and TEACORAC of 3.61). Isomangiferin showed slightly lower antioxidant capacity than mangiferin against DPPH (TEACDPPH of 0.57 vs. 0.62), but higher capacity against ABTS(+) (TEACABTS of 1.82 vs. 1.67) and peroxyl radical (TEACORAC of 4.14 vs. 3.69) than mangiferin. The on-line HPLC-ORAC assay was shown to be more sensitive for radical scavengers, but at the same time less selective for rapid radical scavengers than the DPPH assay.

  14. Diagnostic Accuracy of GeneXpert MTB/RIF Assay in Comparison to Conventional Drug Susceptibility Testing Method for the Diagnosis of Multidrug-Resistant Tuberculosis

    Science.gov (United States)

    Pandey, Pratikshya; Rijal, Komal Raj; Shrestha, Bhawana; Kattel, Sirita; Banjara, Megha Raj; Maharjan, Bhagwan; KC, Rajendra

    2017-01-01

    Xpert MTB/RIF assay is regarded as a great achievement of modern medicine for the rapid diagnosis of multidrug-resistant tuberculosis (MDR-TB). The main purpose of this study was to determine the performance of Xpert MTB/RIF assay compared to conventional drug susceptibility testing (DST) method for the diagnosis of MDR-TB. A comparative cross sectional study was carried out at German-Nepal Tuberculosis Project, Kathmandu, Nepal, from April 2014 to September 2014. A total of 88 culture positive clinical samples (83 pulmonary and 5 extra-pulmonary) received during the study period were analyzed for detection of multidrug-resistant tuberculosis by both GeneXpert MTB/RIF assay and conventional DST method. McNemar chi square test was used to compare the performance of Xpert with that of DST method. A p-value of less than 0.05 was considered as statistically significant. Of total 88 culture positive samples, one was reported as invalid while 2 were found to contain nontuberculous Mycobacteria (NTM). Among remaining 85 Mycobacterium tuberculosis culture positive samples, 69 were found to be MDR-TB positive by both methods. The overall sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of GeneXpert MTB/RIF assay were found to be 98.6%, 100%, 100% and 93.8% respectively. Statistically, there was no significant difference between the diagnostic performance of Xpert and conventional DST method for detection of MDR-TB. GeneXpert MTB/RIF assay was found to be highly sensitive, specific and comparable to gold standard conventional DST method for the diagnosis of MDR-TB. PMID:28081227

  15. Interlaboratory comparison of three microbial source tracking quantitative polymerase chain reaction (qPCR) assays from fecal-source and environmental samples

    Science.gov (United States)

    Stelzer, Erin A.; Strickler, Kriston M.; Schill, William B.

    2012-01-01

    During summer and early fall 2010, 15 river samples and 6 fecal-source samples were collected in West Virginia. These samples were analyzed by three laboratories for three microbial source tracking (MST) markers: AllBac, a general fecal indicator; BacHum, a human-associated fecal indicator; and BoBac, a ruminant-associated fecal indicator. MST markers were analyzed by means of the quantitative polymerase chain reaction (qPCR) method. The aim was to assess interlaboratory precision when the three laboratories used the same MST marker and shared deoxyribonucleic acid (DNA) extracts of the samples, but different equipment, reagents, and analyst experience levels. The term assay refers to both the markers and the procedure differences listed above. Interlaboratory precision was best for all three MST assays when using the geometric mean absolute relative percent difference (ARPD) and Friedman's statistical test as a measure of interlaboratory precision. Adjustment factors (one for each MST assay) were calculated using results from fecal-source samples analyzed by all three laboratories and applied retrospectively to sample concentrations to account for differences in qPCR results among labs using different standards and procedures. Following the application of adjustment factors to qPCR results, ARPDs were lower; however, statistically significant differences between labs were still observed for the BacHum and BoBac assays. This was a small study and two of the MST assays had 52 percent of samples with concentrations at or below the limit of accurate quantification; hence, more testing could be done to determine if the adjustment factors would work better if the majority of sample concentrations were above the quantification limit.

  16. Comparison of Simplexa universal direct PCR with cytotoxicity assay for diagnosis of Clostridium difficile infection: performance, cost, and correlation with disease.

    Science.gov (United States)

    Landry, Marie L; Ferguson, David; Topal, Jeffrey

    2014-01-01

    Simplexa Clostridium difficile universal direct PCR, a real-time PCR assay for the detection of the C. difficile toxin B (tcdB) gene using the 3M integrated cycler, was compared with a two-step algorithm which includes the C. Diff Chek-60 glutamate dehydrogenase (GDH) antigen assay followed by cytotoxin neutralization. Three hundred forty-two liquid or semisolid stools submitted for diagnostic C. difficile testing, 171 GDH antigen positive and 171 GDH antigen negative, were selected for the study. All samples were tested by the C. Diff Chek-60 GDH antigen assay, cytotoxin neutralization, and Simplexa direct PCR. Of 171 GDH-positive samples, 4 were excluded (from patients on therapy or from whom duplicate samples were obtained) and 88 were determined to be true positives for toxigenic C. difficile. Of the 88, 67 (76.1%) were positive by the two-step method and 86 (97.7%) were positive by PCR. Seventy-nine were positive by the GDH antigen assay only. Of 171 GDH antigen-negative samples, none were positive by PCR. One antigen-negative sample positive by the cytotoxin assay only was deemed a false positive based on chart review. Simplexa C. difficile universal direct PCR was significantly more sensitive for detecting toxigenic C. difficile bacteria than cytotoxin neutralization (P = 0.0002). However, most PCR-positive/cytotoxin-negative patients did not have clear C. difficile disease. The estimated cost avoidance provided by a more rapid molecular diagnosis was outweighed by the cost of isolating and treating PCR-positive/cytotoxin-negative patients. The costs, clinical consequences, and impact on nosocomial transmission of treating and/or isolating patients positive for toxigenic C. difficile by PCR but negative for in vivo toxin production merit further study.

  17. [Comparison of remnant lipoprotein-cholesterol measurements: the immune adsorption method (RLP-C) and the direct assay with detergent (RemL-C)].

    Science.gov (United States)

    Hihara, Mari; Sato, Itsuko; Hayashi, Fujio; Sugiyama, Daisuke; Kawano, Seiji; Fujioka, Yoshio; Ishikawa, Yuichi; Kumagai, Shunichi

    2009-01-01

    Elevation of serum remnant lipoprotein concentration is an emerging risk factor for coronary artery disease. An immunoseparation procedure for remnant-like particle cholesterol(RLP-C) has been evaluated extensively in recent years. In addition, a new detergent-based method has been developed and applied to automated analyzer as "MetaboLead RemL-C" (RemL-C, KYOWA MEDEX CO., LTD.). Then, we compared the concentrations of remnant lipoproteins as RemL-C with those as RLP-C in various conditions. RemL-C assay was intra-assay-reproducible (n=20, CVs: 0.6-2.2%), and reproducible for 2 days in the refrigeration and for 8 hours in room temperature. This assay was also inter-assay-reproducible (during 5 days in the deep freezing, CVs: 1.6-3.0%). The available range for RemL-C assay was between 0.09 and 121.1 mg/dl. There were no detectable interferences from hemoglobin, free/conjugated bilirubin, chyle, and Intrafat. However, heparin influenced the titer of RemL-C concentrations. Correlation of values between RLP-C and RemL-C in 123 samples was excellent (r=0.924, p<0.001). However, different responses to intermediate lipoprotein fraction derived from a patients with type III hyperlipidemia were observed. In conclusion, RemL-C and RLP-C measurements may have a similar clinical significance. Differences in sensitivity for intermediate lipoprotein fraction between both methods may exist.

  18. Research on Comparison between Chinese and Western Medicine Diagnosis and Treatment of Chest Stuffiness and Pains%胸痹心痛中西医诊断与治疗研究对比

    Institute of Scientific and Technical Information of China (English)

    张敏; 邹昕

    2016-01-01

    通过对胸痹心痛(冠心病)在中医和西医中的病因、诊断及治疗方法研究进行综述并对比,结合目前实际治疗现状提出中西医结合共同诊断治疗的建议,为理清胸痹心痛的历史和开发新治疗药物及医疗方法提供依据。%In this paper,it is evident that the comparison between traditional Chinese medicine and western medicine etiology,diagnosis and treatment for obstruction of heartache (CHD)are reviewed and compared.Then,the paper proposes status quo in combining common diagnosis of traditional Chi-nese medicine with western medicine treatment,and provides the basis for development of new drugs and medical methods.

  19. Comparison of PCR,DIA and Pathogenicity Assay for Detection of Xanthomonas axonopodis pv.citri,the Causal Agent of Citrus Bacterial Canker Disease

    Institute of Scientific and Technical Information of China (English)

    WANG Zhong-kang; SUN Xian-yun; YIN You-ping; ZHOU Chang-yong; XIA Yu-xian

    2004-01-01

    Polymerase chain reaction (PCR) approach based on newly designed primers, JYF5/JYR5, was applied for specific detection of Xanthomonas axonopodis pv.citri(Xac). The efficiency and reliability of PCR method were compared with dot immunobinding assay (DIA) and classical pathogenicity test techniques for detecting suspensions of pure cells of Xac and soaking sap of citrus tissues. Detection sensitivity of PCR was about 4.5 cells or 1.56 pg target DNA per reaction which was higher than that of DIA (ca. 450 cells per dot).These three techniques (PCR assay, DIA and Pathogenecity test) could always detect Xac from symptomatic citrus samples. Different performances were obtained from citrus materials without symptoms, and the positive detection frequency was PCR, DIA and pathogenicity test.

  20. Comparison of Three Enzyme-Linked Immunosorbent Assays for Detection of Immunoglobulin G Antibodies to Tetanus Toxoid with Reference Standards and the Impact on Clinical Practice▿

    OpenAIRE

    van Hoeven, Karen H.; Dale, Connie; Foster, Phil; Body, Barbara

    2008-01-01

    Accurate determination of the concentrations of immunoglobulin G (IgG) antibody to tetanus toxoid is important in order to evaluate the immunogenicity of tetanus toxoid vaccines, determine immune competence in individual patients, and measure the prevalence of immunity in populations. The performance of three commercially available enzyme-linked immunosorbent assays (ELISAs) for IgG antibodies to tetanus toxoid were evaluated. Serially diluted NIBSC 76/589 and TE-3 human tetanus IgG immunoglo...

  1. Comparison of three real-time PCR assays for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae in young pregnant women.

    Science.gov (United States)

    Peuchant, Olivia; de Diego, Sabrina; Le Roy, Chloé; Frantz-Blancpain, Sandrine; Hocké, Claude; Bébéar, Cécile; de Barbeyrac, Bertille

    2015-12-01

    We compared 3 commercial real-time PCR assays, the Abbott RealTime CT/NG, the cobas® 4800 CT/NG, and the Cepheid Xpert® CT/NG, for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae in vaginal swabs collected prospectively from pregnant women aged gonorrhoeae, the overall agreement was 100%. All kits allowed prompt and specific results for C. trachomatis and N. gonorrhoeae in young pregnant women.

  2. Longitudinal study of the detection of Bluetongue virus in bull semen and comparison of real-time polymerase chain reaction assays.

    Science.gov (United States)

    Gu, Xingnian; Davis, Rodney J; Walsh, Susan J; Melville, Lorna F; Kirkland, Peter D

    2014-01-01

    Infection with Bluetongue virus (BTV) is a significant impediment to the global movement of bovine semen. Repeat testing of blood from donor animals is specified in the World Organization for Animal Health (OIE) Manual for the export of semen from regions where BTV may be present. Screening of blood or semen samples has usually been carried out by virus isolation (VI) either by inoculation of chicken embryos followed by passage onto insect and mammalian cell cultures or in vivo inoculation of sheep followed by serology to detect seroconversion. Direct testing of semen for BTV would enable earlier release of semen samples and avoid repeat testing of the donor, as well as provide an option for releasing batches of semen that were collected without certification of the donor. Quantitative (real-time) reverse transcription polymerase chain reaction (qRT-PCR) assays overcome most of the limitations of other methods and have the potential to provide higher sensitivity. The present study compared 5 qRT-PCR assays, including 2 commercially available kits, for the detection of BTV in semen serially collected from 8 bulls over a period of 90 days after experimental infection. The results of the study show that at least one of the qRT-PCR assays is extremely reproducible and has both very high sensitivity and specificity to reliably detect all available serotypes. The preferred qRT-PCR gave consistently superior results to VI, sheep inoculation, and conventional RT-PCR. Therefore, the assay can be recommended for the screening of bovine semen for freedom from BTV.

  3. Development and evaluation of an ELIME assay to reveal the presence of Salmonella in irrigation water: Comparison with Real-Time PCR and the Standard Culture Method.

    Science.gov (United States)

    Volpe, G; Delibato, E; Fabiani, L; Pucci, E; Piermarini, S; D'Angelo, A; Capuano, F; De Medici, D; Palleschi, G

    2016-01-01

    A reliable, low-cost and easy-to-use ELIME (Enzyme-Linked-Immuno-Magnetic-Electrochemical) assay for detection of Salmonella enterica in irrigation water is presented. Magnetic beads (MBs), coupled to a strip of eight-magnetized screen-printed electrodes localized at the bottom of eight wells (8-well/SPE strip), effectively supported a sandwich immunological chain. Enzymatic by-product is quickly measured by chronoamperometry, using a portable instrument. With the goal of developing a method able to detect a wide range of Salmonella serotypes, including S. Napoli and S. Thompson strains responsible for various community alerts, different kinds of MBs, antibodies and blocking agents were tested. The final system employs MBs coated with a broad reactivity monoclonal antibody anti-salmonella and blocked with dry milk. For a simple and rapid assay these two steps were performed in a preliminary phase, while the two sequential incubations for the immuno-recognition events were merged in a single step of 1h. In parallel a Real-Time PCR (RTi-PCR) method, based on a specific locked nucleic acid (LNA) fluorescent probe and an internal amplification control (IAC), was carried out. The selectivity of the ELIME and RTi-PCR assays was proved by inclusivity and exclusivity tests performed analyzing different Salmonella serotypes and non-target microorganisms, most commonly isolated from environmental sources. Furthermore, both methods were applied to experimentally and not experimentally contaminated irrigation water samples. Results confirmed by the ISO culture method, demonstrated the effectiveness of ELIME and RTi-PCR assays to detect a low number of salmonella cells (1-10 CFU/L) reducing drastically the long analysis time usually required to reveal this pathogen.

  4. Screening complex effluents for estrogenic activity with the T47D-KBluc cell bioassay: assay optimization and comparison with in vivo responses in fish.

    Science.gov (United States)

    Wehmas, Leah C; Cavallin, Jenna E; Durhan, Elizabeth J; Kahl, Michael D; Martinovic, Dalma; Mayasich, Joe; Tuominen, Tim; Villeneuve, Daniel L; Ankley, Gerald T

    2011-02-01

    Wastewater treatment plant (WWTP) effluents can contain estrogenic chemicals, which potentially disrupt fish reproduction and development. The current study focused on the use of an estrogen-responsive in vitro cell bioassay (T47D-KBluc), to quantify total estrogenicity of WWTP effluents. We tested a novel sample preparation method for the T47D-KBluc assay, using powdered media prepared with direct effluent. Results of the T47D-KBluc assay were compared with the induction of estrogen receptor-regulated gene transcription in male fathead minnows (Pimephales promelas) exposed to the same effluents. Effluent samples for the paired studies were collected over the course of three months. According to the T47D-KBluc assay, the effluent estrogenicity ranged from 1.13 to 2.00 ng 17β-estradiol (E2) equivalents/L. Corresponding in vivo studies exposing male fathead minnows to 0, 10, 50, and 100% effluent dilutions demonstrated that exposure to 100% effluent significantly increased hepatic vitellogenin (VTG) and estrogen receptor α subunit transcripts relative to controls. The induction was also significant in males exposed to 250 ng E2/L or 100 ng E2/L. The in vitro and in vivo results support the conclusion that the effluent contains significant estrogenic activity, but there was a discrepancy between in vitro- and in vivo-based E2 equivalent estimates. Our results suggest that the direct effluent preparation method for the T47D-KBluc assay is a reasonable approach to estimate the estrogenicity of wastewater effluent.

  5. Flow Cytometric Immunobead Assay for Detection of BCR-ABL1 Fusion Proteins in Chronic Myleoid Leukemia: Comparison with FISH and PCR Techniques.

    Directory of Open Access Journals (Sweden)

    Anna Grazia Recchia

    Full Text Available Chronic Myeloid Leukemia (CML is characterized by a balanced translocation juxtaposing the Abelson (ABL and breakpoint cluster region (BCR genes. The resulting BCR-ABL1 oncogene leads to increased proliferation and survival of leukemic cells. Successful treatment of CML has been accompanied by steady improvements in our capacity to accurately and sensitively monitor therapy response. Currently, measurement of BCR-ABL1 mRNA transcript levels by real-time quantitative PCR (RQ-PCR defines critical response endpoints. An antibody-based technique for BCR-ABL1 protein recognition could be an attractive alternative to RQ-PCR. To date, there have been no studies evaluating whether flow-cytometry based assays could be of clinical utility in evaluating residual disease in CML patients. Here we describe a flow-cytometry assay that detects the presence of BCR-ABL1 fusion proteins in CML lysates to determine the applicability, reliability, and specificity of this method for both diagnosis and monitoring of CML patients for initial response to therapy. We show that: i CML can be properly diagnosed at onset, (ii follow-up assessments show detectable fusion protein (i.e. relative mean fluorescent intensity, rMFI%>1 when BCR-ABL1IS transcripts are between 1-10%, and (iii rMFI% levels predict CCyR as defined by FISH analysis. Overall, the FCBA assay is a rapid technique, fully translatable to the routine management of CML patients.

  6. Flow Cytometric Immunobead Assay for Detection of BCR-ABL1 Fusion Proteins in Chronic Myleoid Leukemia: Comparison with FISH and PCR Techniques.

    Science.gov (United States)

    Recchia, Anna Grazia; Caruso, Nadia; Bossio, Sabrina; Pellicanò, Mariavaleria; De Stefano, Laura; Franzese, Stefania; Palummo, Angela; Abbadessa, Vincenzo; Lucia, Eugenio; Gentile, Massimo; Vigna, Ernesto; Caracciolo, Clementina; Agostino, Antolino; Galimberti, Sara; Levato, Luciano; Stagno, Fabio; Molica, Stefano; Martino, Bruno; Vigneri, Paolo; Di Raimondo, Francesco; Morabito, Fortunato

    2015-01-01

    Chronic Myeloid Leukemia (CML) is characterized by a balanced translocation juxtaposing the Abelson (ABL) and breakpoint cluster region (BCR) genes. The resulting BCR-ABL1 oncogene leads to increased proliferation and survival of leukemic cells. Successful treatment of CML has been accompanied by steady improvements in our capacity to accurately and sensitively monitor therapy response. Currently, measurement of BCR-ABL1 mRNA transcript levels by real-time quantitative PCR (RQ-PCR) defines critical response endpoints. An antibody-based technique for BCR-ABL1 protein recognition could be an attractive alternative to RQ-PCR. To date, there have been no studies evaluating whether flow-cytometry based assays could be of clinical utility in evaluating residual disease in CML patients. Here we describe a flow-cytometry assay that detects the presence of BCR-ABL1 fusion proteins in CML lysates to determine the applicability, reliability, and specificity of this method for both diagnosis and monitoring of CML patients for initial response to therapy. We show that: i) CML can be properly diagnosed at onset, (ii) follow-up assessments show detectable fusion protein (i.e. relative mean fluorescent intensity, rMFI%>1) when BCR-ABL1IS transcripts are between 1-10%, and (iii) rMFI% levels predict CCyR as defined by FISH analysis. Overall, the FCBA assay is a rapid technique, fully translatable to the routine management of CML patients.

  7. A Comparison of Fresh Frozen vs. Formalin-Fixed, Paraffin-Embedded Specimens of Canine Mammary Tumors via Branched-DNA Assay

    Directory of Open Access Journals (Sweden)

    Florenza Lüder Ripoli

    2016-05-01

    Full Text Available Mammary neoplasms are the tumors most affecting female dogs and women. Formalin-fixed, paraffin-embedded (FFPE tissues are an invaluable source of archived biological material. Fresh frozen (FF tissue is considered ideal for gene expression analysis. However, strategies based on FFPE material offer several advantages. Branched-DNA assays permit a reliable and fast workflow when analyzing gene expression. The aim of this study was to assess the comparability of the branched-DNA assay when analyzing certain gene expression patterns between FF and FFPE samples in canine mammary tumors. RNA was isolated from 109 FFPE samples and from 93 FF samples of different canine mammary tissues. Sixteen (16 target genes (Tp53; Myc; HMGA1; Pik3ca; Mcl1; MAPK3; FOXO3; PTEN; GATA4; PFDN5; HMGB1; MAPK1; BRCA2; BRCA1; HMGA2; and Her2 were analyzed via branched-DNA assay (b-DNA. ACTB, GAPDH, and HPRT1 were used as data normalizers. Overall, the relative gene expression of the two different origins of samples showed an agreement of 63%. Still, care should be taken, as FFPE specimens showed lower expression of the analyzed targets when compared to FF samples. The fact that the gene expression in FFPE proved to be lower than in FF specimens is likely to have been caused by the effect of storage time. ACTB had the best performance as a data normalizer.

  8. Comparison of Laboratory-Developed and Commercial Monoclonal Antibody-Based Sandwich Enzyme-Linked Immunosorbent Assays for Almond (Prunus dulcis) Detection and Quantification.

    Science.gov (United States)

    Liu, Changqi; Chhabra, Guneet S; Zhao, Jing; Zaffran, Valerie D; Gupta, Sahil; Roux, Kenneth H; Gradziel, Thomas M; Sathe, Shridhar K

    2017-09-04

    A commercially available monoclonal antibody (mAb)-based direct sandwich enzyme-linked immunosorbent assay (ELISA) kit (BioFront Technologies, Tallahassee, Fla., U.S.A.) was compared with an in-house developed mAb 4C10-based ELISA for almond detection. The assays were comparable in sensitivity (limit of detection < 1 ppm full fat almond, limit of quantification < 5 ppm full fat almond), specificity (no cross-reactivity with 156 tested foods at a concentration of 100000 ppm whole sample), and reproducibility (intra- and interassay variability < 15% CV). The target antigens were stable and detectable in whole almond seeds subjected to autoclaving, blanching, frying, microwaving, and dry roasting. The almond recovery ranges for spiked food matrices were 84.3% to 124.6% for 4C10 ELISA and 81.2% to 127.4% for MonoTrace ELISA. The almond recovery ranges for commercial and laboratory prepared foods with declared/known almond amount were 30.9% to 161.2% for 4C10 ELISA and 38.1% to 207.6% for MonoTrace ELISA. Neither assay registered any false-positive or negative results among the tested commercial and laboratory prepared samples. © 2017 Institute of Food Technologists®.

  9. Comparison of Procleix Ultrio Elite and Procleix Ultrio NAT Assays for Screening of Transfusion Transmitted Infections among Blood Donors in India

    Directory of Open Access Journals (Sweden)

    Rahul Chaurasia

    2016-01-01

    Full Text Available Background. Introduction of nucleic acid testing (NAT has helped in decreasing window period donations, resulting in increased safety of blood supplies. NAT combines the advantages of direct and highly sequence-specific detection of viral genomes. We analysed the performance of newer Procleix Ultrio Elite (PUE and Procleix Ultrio assay (PUA for the screening of the viral markers in our donor population. Material and Methods. 10,015 donor samples were screened by routine immunoassays and both versions of NAT. NAT yields detected were subjected to viral load estimation and to other serological markers. Results. A total of 21 NAT yields were detected; three were positive by both NAT systems, whereas 18 samples were reactive by PUE only. NAT yields include 18 HBV and 3 HCV yields, of which 17 HBV yields were occult infections and 1 was window period (WP infection. All 3 HCV yields were WP infections. No HIV-1/HIV-2 yield was found. Conclusion. Efficient target capture chemistry in the new TMA assay version significantly improved sensitivity. NAT is superior to serological immunoassays for screening of the viral markers; and the efficient target capture system in the newer TMA assay, namely, the PUE system, has significantly improved sensitivity over the earlier versions.

  10. Comparison of Fluorescent Microspheres and Colloidal Gold as Labels in Lateral Flow Immunochromatographic Assays for the Detection of T-2 Toxin

    Directory of Open Access Journals (Sweden)

    Xiya Zhang

    2015-12-01

    Full Text Available A new highly specific and sensitive monoclonal antibody (MAb to T-2 toxin (T-2 was produced, providing an IC50 value of 1.02 ng/mL and negligible cross-reactivity (CR to other related mycotoxins. Based on the new MAb, a lateral-flow immunochromatographic assay (LFIA using colloidal gold (CG and fluorescent microspheres (FMs as labels was proposed for T-2. Under the optimized conditions, in rapid qualitative assay, the cut-off values of the CG-LFIA were 400 μg/kg in rice and 50 μg/L in fresh milk, and the cut-off values of the FMs-LFIA were 100 μg/kg in both rice and chicken feed. For the quantitative assay with the FMs-LFIA, the limit of detection (LOD were 0.23 μg/kg and 0.41 μg/kg in rice and chicken feed, respectively, and the average recoveries ranged from 80.2% to 100.8% with the coefficient of variation (CV below 10.8%. In addition, we found that the CG-LFIA could tolerate the matrix effect of fresh milk better than the FMs-LFIA, while the FMs-LFIA could tolerate the matrix effect of chicken feed better than CG-LFIA under the same experimental conditions. These results provide a certain reference for the selection of appropriate labels to establish a rapid LFIA in various biological samples.

  11. Seabird Colonies in Western Greenland

    DEFF Research Database (Denmark)

    Boertmann, D.; Mosbech, A.; Falk, K.;

    surveys of seabird colonies are needed, due to a lack of information or because the present information probably is outdated. The most immediate threats to the colonial seabirds in western Greenland during the breeding time is hunting and egging. Oil pollution is a minor threat to-day, but will increase...... if offshore areas with oil potential are explored and developed. Tab. 6 gives an overview of each species sensitivity to oil spills and the capacity to recover, as well as a comparison of the western Greenland population numbers to the North Atlantic population numbers. The most significant western Greenland...

  12. IgA anti-Actin antibodies in children with celiac disease: comparison of immunofluorescence with Elisa assay in predicting severe intestinal damage

    Directory of Open Access Journals (Sweden)

    Mora Stefano

    2010-03-01

    Full Text Available Abstract Background Previous studies have demonstrated that the presence of serum IgA antibodies against actin filaments (AAA in patients with celiac disease (CD is strongly associated with mucosal damage and severe degrees of villous atrophy. The aims of the present study were (1 to verify the effectiveness of IgA-AAA in newly diagnosed CD patients in a clinical setting (2 to compare the immunofluorescence assay with ELISA assay; (3 to compare the correlation of our IgA anti-tissue transglutaminase antibodies (tTG-Ab class with mucosal intestinal lesions. Methods 90 patients underwent endoscopy and multiple biopsies for suspected CD on the basis of symptoms, in presence of positive tTG-Ab tests. Twenty biopsied and 25 not-biopsied subjects with negative tTG-Ab were tested as control groups. IgA-AAA assays were performed by indirect immunofluorescence using rat epithelial intestinal cells, and by ELISA with a commercial kit. tTG-Ab assay was a radio-binding assay. Intestinal specimens were collected by upper endoscopy and the histological study was done according to the Marsh's classification modified by Oberhuber (M/O. Auto-antibodies assays and histological evaluation have been performed blindly by skilled operators. Results CD diagnosis was confirmed in 82 patients (type I M/O in 2 patients, IIIA in 18 patients, IIIB in 29 patients and IIIC in 33 patients. Two patients with type 1 lesion in presence of positive tTG-Ab and abdominal complaints, started a gluten free diet. The rate of IgA-AAA positivity (sensitivity by IFI and ELISA in histologically proven celiac disease patients, were 5.5% and 25% patients in IIIA, 27.5% and 34.4% patients in IIIB, 78.8% and 75% in IIIC patients, respectively. Patients with normal or nearly normal mucosa, regardless of tTG-Ab status, presented negative IgA-AAA IFI assay. On the other hand, 1 patient with normal mucosa but positive tTG-Ab, also presented positive IgA-AAA ELISA. All healthy non biopsied

  13. Relative performance of Organon kit in comparison to Du Pont for confirmatory serological testing of HIV infection by western blot test in sera from blood donors.

    Science.gov (United States)

    Aggarwal, R K; Chatterjee, R; Chattopadhya, D; Kumari, S

    1992-06-01

    A total of 32 specimens with different categories of reactivity by Du Pont Western Blot kit comprising of specimens showing full spectrum of HIV-I antigen specific bands, 19 specimens showing total absence of bands and four specimens showing non-specific bands (without any interpretative importance) were subjected to Western Blot testing by Organon test. Of the nine specimens showing full spectrum of bands by Du Pont the correlation with Organon kit was 100 per cent based on WHO criteria. Four specimens with non-specific indeterminate band pattern by Du Pont failed to show any band in Organon kit, indicating that latter to be more specific.

  14. Comparison of the conventional multiplex RT-PCR, real time RT-PCR and Luminex xTAG® RVP fast assay for the detection of respiratory viruses.

    Science.gov (United States)

    Choudhary, Manohar L; Anand, Siddharth P; Tikhe, Shamal A; Walimbe, Atul M; Potdar, Varsha A; Chadha, Mandeep S; Mishra, Akhilesh C

    2016-01-01

    Detection of respiratory viruses using polymerase chain reaction (PCR) is sensitive, specific and cost effective, having huge potential for patient management. In this study, the performance of an in-house developed conventional multiplex RT-PCR (mRT-PCR), real time RT-PCR (rtRT-PCR) and Luminex xTAG(®) RVP fast assay (Luminex Diagnostics, Toronto, Canada) for the detection of respiratory viruses was compared. A total 310 respiratory clinical specimens predominantly from pediatric patients, referred for diagnosis of influenza A/H1N1pdm09 from August 2009 to March 2011 were tested to determine performance characteristic of the three methods. A total 193 (62.2%) samples were detected positive for one or more viruses by mRT-PCR, 175 (56.4%) samples by real time monoplex RT-PCR, and 138 (44.5%) samples by xTAG(®) RVP fast assay. The overall sensitivity of mRT-PCR was 96.9% (95% CI: 93.5, 98.8), rtRT-PCR 87.9% (95% CI: 82.5, 92.1) and xTAG(®) RVP fast was 68.3% (95% CI: 61.4, 74.6). Rhinovirus was detected most commonly followed by respiratory syncytial virus group B and influenza A/H1N1pdm09. The monoplex real time RT-PCR and in-house developed mRT-PCR are more sensitive, specific and cost effective than the xTAG(®) RVP fast assay.

  15. Comparison of a Monoclonal Antibody-Blocking Enzyme-Linked Immunoassay and a Strip Immunoblot Assay for Identifying Type-Specific Herpes Simplex Virus Type 2 Serological Responses

    OpenAIRE

    van Doornum, G. J. J.; Slomka, M.J.; Buimer, M; Groen, J.; van den Hoek, J.A.R.; Cairo, I.; Vyse, A.; Brown, D. W G

    2000-01-01

    Detection of herpes simplex virus type 2 (HSV-2)-specific antibodies by a monoclonal antibody (MAb)-blocking enzyme-linked immunoassay (EIA) was compared with detection by a strip immunoblot assay (SIA) in a sexually transmitted disease (STD) clinic population. The study population consisted of 1,683 genitourinary medicine clinic attendees (582 women and 1,101 men). Sera were tested for the presence of HSV-2 antibody by use of the blocking EIA, in which binding of the MAb AP-1 to HSV-2 glycop...

  16. Detection of AmpC Beta-Lactamase in Escherichia coli: Comparison of Three Phenotypic Confirmation Assays and Genetic Analysis▿†

    OpenAIRE

    Peter-Getzlaff, S; Polsfuss, S; Poledica, M.; Hombach, M.; Giger, J.; Böttger, E C; Zbinden, R.; Bloemberg, G. V.

    2011-01-01

    Two mechanisms account for AmpC activity in Escherichia coli, namely, mutations in the ampC promoter and attenuator regions resulting in ampC overexpression and acquisition of plasmid-carried ampC genes. In this study, we analyzed 51 clinical E. coli isolates with reduced susceptibility to amoxicillin-clavulanic acid, piperacillin-tazobactam, or extended-spectrum cephalosporins for the presence of AmpC production. Three phenotypic AmpC confirmation assays (cefoxitin-cloxacillin disk diffusion...

  17. Comparison of Clot-based, Chromogenic, and Fluorescence Assays for Measurement of Factor VIII Inhibitors in the U.S. Hemophilia Inhibitor Research Study

    Science.gov (United States)

    Miller, Connie H.; Rice, Anne S.; Boylan, Brian; Shapiro, Amy D.; Lentz, Steven R.; Wicklund, Brian M.; Kelly, Fiona M.; Soucie, J. Michael

    2015-01-01

    Summary Background Detection and validation of inhibitors (antibodies) to hemophilia treatment products are important for clinical care, evaluation of product safety, and assessment of population trends. Methods Centralized monitoring for factor VIII (FVIII) inhibitors was conducted for patients in the Hemophilia Inhibitor Research Study using a previously reported modified Nijmegen-Bethesda clotting assay (NBA), a chromogenic Bethesda assay (CBA), and a novel fluorescence immunoassay (FLI). Results NBA and CBA were performed on 1005 specimens and FLI on 272 specimens. CBA was negative on 880/883 specimens (99.7%) with Nijmegen-Bethesda units (NBU)<0.5 and positive on 42/42 specimens (100%) with NBU≥2.0 and 43/80 specimens (53.8%) with NBU 0.5–1.9. Among specimens with positive NBA and negative CBA, 58.1% were FLI-negative, 12.9% had evidence of lupus anticoagulant, and 35.5% had non-time-dependent inhibition. CBA and FLI were positive on 72.4% and 100% of 1.0–1.9 NBU specimens and 43.1% and 50.0% of 0.5–0.9 NBU specimens. FLI detected antibodies in 98.0% of CBA-positive and 81.6% of NBA-positive specimens (P=0.004). Among 21 new inhibitors detected by NBA, 5 (23.8%) with 0.7–1.3 NBU did not react in CBA or FLI. Among previously positive patients with 0.5–1.9 NBU, 7/25 (28%) were not CBA or FLI positive. FLI was positive on 36/169 NBU-negative specimens (21.3%). Conclusions FVIII specificity could not be demonstrated by CBA or FLI for 26% of inhibitors of 0.5–1.9 NBU; such results must be interpreted with caution. Low titer inhibitors detected in clot-based assays should always be repeated, with consideration given to evaluating their reactivity with FVIII using more specific assays. PMID:23601690

  18. Detection of toxigenic Clostridium difficile: comparison of the cell culture neutralization, Xpert C. difficile, Xpert C. difficile/Epi, and Illumigene C. difficile assays.

    Science.gov (United States)

    Pancholi, P; Kelly, C; Raczkowski, M; Balada-Llasat, J M

    2012-04-01

    Clostridium difficile is the most important cause of nosocomial diarrhea. Several laboratory techniques are available to detect C. difficile toxins or the genes that encode them in fecal samples. We evaluated the Xpert C. difficile and Xpert C. difficile/Epi (Cepheid, CA) that detect the toxin B gene (tcdB) and tcdB, cdt, and a deletion in tcdC associated with the 027/NAP1/BI strain, respectively, by real-time PCR, and the Illumigene C. difficile (Meridian Bioscience, Inc.) that detects the toxin A gene (tcdA) by loop-mediated isothermal amplification in stool specimens. Toxigenic culture was used as the reference method for discrepant stool specimens. Two hundred prospective and fifty retrospective diarrheal stool specimens were tested simultaneously by the cell cytotoxin neutralization assay (CCNA) and the Xpert C. difficile, Xpert C. difficile/Epi, and Illumigene C. difficile assays. Of the 200 prospective stools tested, 10.5% (n = 23) were determined to be positive by CCNA, 17.5% (n = 35) were determined to be positive by Illumigene C. difficile, and 21.5% (n = 43) were determined to be positive by Xpert C. difficile and Xpert C. difficile/Epi. Of the 50 retrospective stools, previously determined to be positive by CCNA, 94% (n = 47) were determined to be positive by Illumigene C. difficile and 100% (n = 50) were determined to be positive by Xpert C. difficile and Xpert C. difficile/Epi. Of the 11 discrepant results (i.e., negative by Illumigene C. difficile but positive by Xpert C. difficile and Xpert C. difficile/Epi), all were determined to be positive by the toxigenic culture. A total of 21% of the isolates were presumptively identified by the Xpert C. difficile/Epi as the 027/NAP1/BI strain. The Xpert C. difficile and Xpert C. difficile/Epi assays were the most sensitive, rapid, and easy-to use assays for the detection of toxigenic C. difficile in stool specimens.

  19. A review of abortion laws in Western-European countries: A cross-national comparison of legal developments between 1960 and 2010

    NARCIS (Netherlands)

    Levels, M.; Sluiter, R.; Need, A.

    2014-01-01

    The extent to which women have had access to legal abortions has changed dramatically in Western-Europe between 1960 and 2010. In most countries, abortion laws developed from completely banning abortion to allowing its availability on request. Both the timing and the substance of the various legal d

  20. A review of abortion laws in Western-European countries. A cross-national comparison of legal developments between 1960 and 2010

    NARCIS (Netherlands)

    Levels, Mark; Sluiter, Roderick; Need, Ariana

    2014-01-01

    The extent to which women have had access to legal abortions has changed dramatically in Western-Europe between 1960 and 2010. In most countries, abortion laws developed from completely banning abortion to allowing its availability on request. Both the timing and the substance of the various legal d

  1. The Big-Fish-Little-Pond Effect: Generalizability of Social Comparison Processes over Two Age Cohorts from Western, Asian, and Middle Eastern Islamic Countries

    Science.gov (United States)

    Marsh, Herbert W.; Abduljabbar, Adel Salah; Morin, Alexandre J. S.; Parker, Philip; Abdelfattah, Faisal; Nagengast, Benjamin; Abu-Hilal, Maher M.

    2015-01-01

    Extensive support for the seemingly paradoxical negative effects of school- and class-average achievement on academic self-concept (ASC)-the big-fish-little-pond effect (BFLPE)--is based largely on secondary students in Western countries or on cross-cultural Program for International Student Assessment studies. There is little research testing the…

  2. A Comparison of Food Supply from 1984 to 2009 and Degree of Dietary Westernization in Taiwan with Asian Countries and World Continents

    Directory of Open Access Journals (Sweden)

    Cheau-Jane Peng

    2015-01-01

    Full Text Available Objective. To compare quality, quantity, and trends of food supply from 1984 to 2009 and degree of food westernization in Taiwan with Asian countries and world continents by using food balance data. Methods. We compiled data from food balance sheets of Taiwan and Food and Agriculture Organization, including five continents and three most populated countries each in Eastern, Southern, and Southeastern Asia over the period 1984–2009. Quantity of food supply per capita was referenced to Taiwan food guides. The population-weighted means of food supply from Europe, North America, South America, and Australia and New Zealand continents in terms of energy and nutrient distributions, animal/plant sources, and sugar/alcohol contribution were used as indicators of westernization. Trends of food supply per capita of six food groups were plotted, and linear regression was applied to evaluate food changes. Findings. Taiwan’s food supply provided sufficient quantity in food energy, with the lowest cereals/roots supply and rice to wheat ratio, but the highest meat and oil supplies per capita among the 10 studied Asian countries. Taiwan food supply showed the most westernization among these countries. Conclusion. Food supply of Taiwan, although currently sufficient, indicated some security problems and high tendency of diet westernization.

  3. A review of abortion laws in Western-European countries: A cross-national comparison of legal developments between 1960 and 2010

    NARCIS (Netherlands)

    Levels, M.; Sluiter, R.; Need, A.

    2014-01-01

    The extent to which women have had access to legal abortions has changed dramatically in Western-Europe between 1960 and 2010. In most countries, abortion laws developed from completely banning abortion to allowing its availability on request. Both the timing and the substance of the various legal

  4. A review of abortion laws in Western-European countries. A cross-national comparison of legal developments between 1960 and 2010

    NARCIS (Netherlands)

    Levels, Mark; Sluiter, Roderick; Need, Ariana

    2014-01-01

    The extent to which women have had access to legal abortions has changed dramatically in Western-Europe between 1960 and 2010. In most countries, abortion laws developed from completely banning abortion to allowing its availability on request. Both the timing and the substance of the various legal

  5. A review of abortion laws in Western-European countries. A cross-national comparison of legal developments between 1960 and 2010

    NARCIS (Netherlands)

    Levels, Mark; Sluiter, Roderick; Need, Ariana

    2014-01-01

    The extent to which women have had access to legal abortions has changed dramatically in Western-Europe between 1960 and 2010. In most countries, abortion laws developed from completely banning abortion to allowing its availability on request. Both the timing and the substance of the various legal d

  6. Evaluation of enzyme-linked immunosorbent assay in comparison with complement fixation test for the diagnosis of subclinical paratuberculosis in cattle.

    Science.gov (United States)

    Yokomizo, Y; Kishima, M; Mori, Y; Nishimori, K

    1991-08-01

    An enzyme-linked immunosorbent assay (ELISA) was evaluated and compared in parallel with the standard complement fixation test (CFT) for the diagnosis of bovine subclinical paratuberculosis. Bovine sera preabsorbed with the mixture of Mycobacterium phlei and kaolin suspension were assayed for antibody activities to the crude protoplasmic antigen of Mycobacterium paratuberculosis in the ELISA. ELISA antibody titer was expressed as ELISA antibody index (EAI) value: EAI = (At-An)/(Ap-An), where At, Ap and An are the absorbance values of a 1:200 dilution of unknown test sera, a 1:400 dilution of positive control serum, and a 1:200 dilution of negative control serum. An EAI of 0.6 or greater was established as a reasonable cutoff point for a positive antibody titer by ELISA. Of the 156 sera from cattle with subclinical M. paratuberculosis-infection, 106 (67.9%) were positive by ELISA and 41 (26.3%) by CFT. Of the 3,880 sera from cattle in the herds which had no history or evidence of paratuberculosis, 3,875 (99.9%) were negative by ELISA, and 3,787 (97.6%) by CFT. Positive ELISA titers were detectable 1 to 5 months earlier than positive CFT titers in experimentally infected cattle, and 7 to 10 months earlier in naturally infected cattle. These results indicate that the ELISA should replace the CFT as the routine test of choice for the diagnosis of bovine paratuberculosis.

  7. Comparison of Interferon gamma inducible protein-10 and Interferon gamma based QuantiFERON TB Gold assays with tuberculin skin test in HIV infected subjects

    Science.gov (United States)

    Basirudeen, S; Rajasekaran, S; Alamelu, R

    2011-01-01

    We aimed to compare the positivity of the QuantiFERON TB gold in-tube (QFT-IT antigens) specific Interferon gamma (IFN-γ/QFT-IT) and IFN-γ nducible protein-10 (IP-10/QFT-IT) assays with tuberculin skin test (TST) among human immunodeficiency virus (HIV) infected individuals in a TB endemic setting. A total of 180 HIV infected subjects, with no evidence of active TB were recruited. IFN-γ nd IP-10 levels specific to QFT-IT antigens were measured in plasma from QFT-IT tubes. The overall positivity of TST at 5mm cut-off point (19%) was significantly lower when compared to IFN-γ/QFT-IT (38%) and IP-10/QFT-IT (45%) assays. The positivity of IP-10/QFT-IT was significantly higher than IFN-γ/QFT-IT (p=0.038). Indeterminate results for IFN-γ/QFT-IT and IP-10/QFT-IT were more frequent in subjects with CD4 count 100 cells/µl. IFN-γ/QFT-IT (9%) yielded significantly higher number of indeterminate results than IP-10/QFT-IT (5%). The frequency of these responses is higher than the proportion of individuals with positive TST results. However, 6 IFN-γ/QFT-IT or IP-10/QFT-IT negative subjects were positive for TST at 5mm cut-off point. Prospective and prognostic studies are required to clarify the significance of these data. PMID:21996360

  8. Comparison of Luminex xTAG® RVP fast assay and real time RT-PCR for the detection of respiratory viruses in adults with community-acquired pneumonia.

    Science.gov (United States)

    Luchsinger, Vivian; Prades, Yara; Ruiz, Mauricio; Pizarro, Rolando; Rossi, Patricio; Lizama, Luis; Garmendia, María Luisa; Meza, Angela; Larrañaga, Carmen; Avendaño, Luis F

    2016-07-01

    Community-acquired pneumonia (CAP) is the third cause of death worldwide. Viruses are frequently detected in adult CAP. Highly sensitive diagnostic techniques should be used due to poor viral shedding. Different sampling methods can affect viral detection, being necessary to establish the optimal type of sample for identifying respiratory viruses in adults. The detection rates of respiratory viruses by Luminex xTAG® RVP fast assay, real time RT-PCR (rtRT-PCR) (Sacace®), and immunofluorescence assay (IFA) in adult CAP were performed in nasopharyngeal swabs (NPS) and aspirates (NPA) from 179 hospitalized adults. Positivity was 47.5% for Luminex®, 42.5% for rtRT-PCR (P = 0.3), and 2.7% for IFA (2.7%) (P viruses in 112 NPA and 35 (34.3%) and 31 (30.4%) in 102 NPS, respectively (P viruses in CAP adults. Both molecular techniques yielded better results with nasopharyngeal aspirate than swabs.

  9. Evaluation of an enzyme-linked immunosorbent assay (ELISA) for the detection of Campylobacter jejuni antibodies, and comparison with a complement fixation test (CFT).

    Science.gov (United States)

    Oosterom, J; den Uyl, C H; Bänffer, J R; Lauwers, S; Huisman, J; Busschbach, A E; Poelma, F G; Bellemans, R

    1985-01-01

    An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of total anti-Campylobacter immunoglobulins in human sera. In this assay disintegrated Campylobacter bacteria were used as the antigen. Absorption tests including other possibly enteropathogenic bacterial species showed that the ELISA system displayed a high immunological specificity for Campylobacter. Using this ELISA it was found that in about 80% of Campylobacter patients these Campylobacter antibodies are produced to almost maximal levels within 8 days after onset of disease, and that they may persist for at least 4 months. Indeed, Campylobacter antibodies were demonstrated at low levels in a large number of control sera. However, accepting an antibody titre of 1:640 as indicative of Campylobacter infection, the statistical sensitivity of the ELISA system was 77% and the specificity 95%. In an epidemiological survey a high association was demonstrated between the severity of Campylobacter-related symptoms and antibody titre values. Assessment of Campylobacter antibody titres by means of this ELISA and by a complement fixation test in 92 sera from index patients and contacts with and without symptoms showed a high association of results.

  10. Comparison of fluorescence polarization assay with Rose Bengal (RB test and complement fixation tests for the diagnosis of buffalo (Bubalus bubalis brucellosis in a high-prevalence area

    Directory of Open Access Journals (Sweden)

    G. Iovane

    2010-02-01

    Full Text Available The fluorescence polarisation assay (FPA was evaluated for the serological diagnosis of bovine brucellosis in buffalo (Bubalus Bubalis in southern Italy. This assay uses O-polysaccharide prepared from Brucella Abortus lipopolysaccharide conjugated with fluorescein isothiocyanate as a tracer. It has many methodological advantages over older, more established tests and can be performed in short time. To measure the fluorescence polarization, a Tecan GENios Pro (Prionics fluorescence-polarization analyzer was used with the procedure described by Nielsen et al. 1996. A cut-off value of 117 millipolarization (mP units was used for testing 912 buffalo sera from Campania Region (526 positive sera and 386 negative sera according to the complement fixation test; CFT. All samples were tested with the Rose Bengal plate (RB. Sensitivity and specificity (Sn for RB were 84.0% and 87.8% and for FPA were 92.6% and 88.9 percent. The FPA gave a kappa coefficient of agreement with respect to CFT of 0.755, while RB (relative to the CFT gave coefficients of 0,715. Finally, ROC analysis suggested a cut-off value which did not agree with the one recommended in the test procedure for cow.

  11. Comparison of a frozen human foreskin fibroblast cell assay to an enzyme immunoassay and toxigenic culture for the detection of toxigenic Clostridium difficile☆☆☆★

    Science.gov (United States)

    Strachan, Alastair J.; Evans, Natalie E.; Williams, O. Martin; Spencer, Robert C.; Greenwood, Rosemary; Probert, Chris J.

    2013-01-01

    This study set out to validate the Hs27 ReadyCell assay (RCCNA) as an alternative CCNA method compared against a commonly used commercial enzyme immunoassay (EIA) method and toxigenic culture (TC) reference standard. A total of 860 samples were identified from those submitted to the Health Protection Agency microbiology laboratories over a 30-week period. RCCNA performed much better than EIA when using TC as a gold standard, with sensitivities of 90.8% versus 78.6% and positive predictive value of 87.3% to 81.9%, respectively. The Hs27 Human Foreskin Fibroblast ReadyCells are an easy-to-use and a sensitive CCNA method for the detection of toxigenic Clostridium difficile directly from stool. A turnaround time of up to 48 h for a negative result and possible need for repeat testing make it an unsuitable method to be used in most clinical laboratory setting. PMID:23107315

  12. Comparison of a frozen human foreskin fibroblast cell assay to an enzyme immunoassay and toxigenic culture for the detection of toxigenic Clostridium difficile.

    Science.gov (United States)

    Strachan, Alastair J; Evans, Natalie E; Williams, O Martin; Spencer, Robert C; Greenwood, Rosemary; Probert, Chris J

    2013-01-01

    This study set out to validate the Hs27 ReadyCell assay (RCCNA) as an alternative CCNA method compared against a commonly used commercial enzyme immunoassay (EIA) method and toxigenic culture (TC) reference standard. A total of 860 samples were identified from those submitted to the Health Protection Agency microbiology laboratories over a 30-week period. RCCNA performed much better than EIA when using TC as a gold standard, with sensitivities of 90.8% versus 78.6% and positive predictive value of 87.3% to 81.9%, respectively. The Hs27 Human Foreskin Fibroblast ReadyCells are an easy-to-use and a sensitive CCNA method for the detection of toxigenic Clostridium difficile directly from stool. A turnaround time of up to 48 h for a negative result and possible need for repeat testing make it an unsuitable method to be used in most clinical laboratory setting.

  13. IS711-based real-time PCR assay as a tool for detection of Brucella spp. in wild boars and comparison with bacterial isolation and serology

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    Miserez Raymond

    2009-07-01

    Full Text Available Abstract Background Control of brucellosis in livestock, wildlife and humans depends on the reliability of the methods used for detection and identification of bacteria. In the present study, we describe the evaluation of the recently established real-time PCR assay based on the Brucella-specific insertion sequence IS711 with blood samples from 199 wild boars (first group of animals and tissue samples from 53 wild boars (second group of animals collected in Switzerland. Results from IS711 real-time PCR were compared to those obtained by bacterial isolation, Rose Bengal Test (RBT, competitive ELISA (c-ELISA and indirect ELISA (i-ELISA. Results In the first group of animals, IS711 real-time PCR detected infection in 11.1% (16/144 of wild boars that were serologically negative. Serological tests showed different sensitivities [RBT 15.6%, c-ELISA 7.5% and i-ELISA 5.5%] and only 2% of blood samples were positive with all three tests, which makes interpretation of the serological results very difficult. Regarding the second group of animals, the IS711 real-time PCR detected infection in 26% of animals, while Brucella spp. could be isolated from tissues of only 9.4% of the animals. Conclusion The results presented here indicate that IS711 real-time PCR assay is a specific and sensitive tool for detection of Brucella spp. infections in wild boars. For this reason, we propose the employment of IS711 real-time PCR as a complementary tool in brucellosis screening programs and for confirmation of diagnosis in doubtful cases.

  14. Comparison of Three Enzyme-Linked Immunosorbent Assays for Detection of Immunoglobulin G Antibodies to Tetanus Toxoid with Reference Standards and the Impact on Clinical Practice▿

    Science.gov (United States)

    van Hoeven, Karen H.; Dale, Connie; Foster, Phil; Body, Barbara

    2008-01-01

    Accurate determination of the concentrations of immunoglobulin G (IgG) antibody to tetanus toxoid is important in order to evaluate the immunogenicity of tetanus toxoid vaccines, determine immune competence in individual patients, and measure the prevalence of immunity in populations. The performance of three commercially available enzyme-linked immunosorbent assays (ELISAs) for IgG antibodies to tetanus toxoid were evaluated. Serially diluted NIBSC 76/589 and TE-3 human tetanus IgG immunoglobulin international reference standards were analyzed in quadruplicate using ELISAs manufactured by The Binding Site, Inc. (VaccZyme); Scimedx; and Euroimmun. In addition, IgG antibodies to tetanus toxoid were measured in 83 deidentified serum specimens using each manufacturer's ELISA. Each ELISA provided linear results when evaluated with the reference preparations. The Binding Site ELISA provided results that closely corresponded to the reference preparations (y = 1.09x − 0.08), whereas the Scimedx ELISA gave results that were consistently lower (y = 0.21x − 0.07) and the Euroimmun ELISA gave results that were consistently higher (y = 1.5x + 0.30) than the reference preparation concentrations. Using the recommended cutoff for each ELISA (<0.10 IU/ml), the overall agreement of all of the ELISA methods was 78%. Three of eighty-three (3.6%) human serum samples demonstrated inadequate immunity with all three assays. The Binding Site ELISA yielded nonprotective antibody concentrations in only these 3 samples, whereas 19 samples (22.9%) according to the Scimedx ELISA and 6 samples (7.2%) according to the Euroimmun ELISA demonstrated nonprotective concentrations. The performance characteristics of ELISAs for tetanus immunoglobulin titers were manufacturer dependent, and the differences translated into important disparities in reported results. PMID:18845832

  15. Comparison of three enzyme-linked immunosorbent assays for detection of immunoglobulin g antibodies to tetanus toxoid with reference standards and the impact on clinical practice.

    Science.gov (United States)

    van Hoeven, Karen H; Dale, Connie; Foster, Phil; Body, Barbara

    2008-12-01

    Accurate determination of the concentrations of immunoglobulin G (IgG) antibody to tetanus toxoid is important in order to evaluate the immunogenicity of tetanus toxoid vaccines, determine immune competence in individual patients, and measure the prevalence of immunity in populations. The performance of three commercially available enzyme-linked immunosorbent assays (ELISAs) for IgG antibodies to tetanus toxoid were evaluated. Serially diluted NIBSC 76/589 and TE-3 human tetanus IgG immunoglobulin international reference standards were analyzed in quadruplicate using ELISAs manufactured by The Binding Site, Inc. (VaccZyme); Scimedx; and Euroimmun. In addition, IgG antibodies to tetanus toxoid were measured in 83 deidentified serum specimens using each manufacturer's ELISA. Each ELISA provided linear results when evaluated with the reference preparations. The Binding Site ELISA provided results that closely corresponded to the reference preparations (y=1.09x-0.08), whereas the Scimedx ELISA gave results that were consistently lower (y=0.21x-0.07) and the Euroimmun ELISA gave results that were consistently higher (y=1.5x+0.30) than the reference preparation concentrations. Using the recommended cutoff for each ELISA (<0.10 IU/ml), the overall agreement of all of the ELISA methods was 78%. Three of eighty-three (3.6%) human serum samples demonstrated inadequate immunity with all three assays. The Binding Site ELISA yielded nonprotective antibody concentrations in only these 3 samples, whereas 19 samples (22.9%) according to the Scimedx ELISA and 6 samples (7.2%) according to the Euroimmun ELISA demonstrated nonprotective concentrations. The performance characteristics of ELISAs for tetanus immunoglobulin titers were manufacturer dependent, and the differences translated into important disparities in reported results.

  16. Rapid identification of ascomycetous yeasts from clinical specimens by a molecular method based on flow cytometry and comparison with identifications from phenotypic assays.

    Science.gov (United States)

    Page, Brent T; Shields, Christine E; Merz, William G; Kurtzman, Cletus P

    2006-09-01

    This study was designed to compare the identification of ascomycetous yeasts recovered from clinical specimens by using phenotypic assays (PA) and a molecular flow cytometric (FC) method. Large-subunit rRNA domains 1 and 2 (D1/D2) gene sequence analysis was also performed and served as the reference for correct strain identification. A panel of 88 clinical isolates was tested that included representatives of nine commonly encountered species and six infrequently encountered species. The PA included germ tube production, fermentation of seven carbohydrates, morphology on corn meal agar, urease and phenoloxidase activities, and carbohydrate assimilation tests when needed. The FC method (Luminex) employed species-specific oligonucleotides attached to polystyrene beads, which were hybridized with D1/D2 amplicons from the unidentified isolates. The PA identified 81 of 88 strains correctly but misidentified 4 of Candida dubliniensis, 1 of C. bovina, 1 of C. palmioleophila, and 1 of C. bracarensis. The FC method correctly identified 79 of 88 strains and did not misidentify any isolate but did not identify nine isolates because oligonucleotide probes were not available in the current library. The FC assay takes approximately 5 h, whereas the PA takes from 2 h to 5 days for identification. In conclusion, PA did well with the commonly encountered species, was not accurate for uncommon species, and takes significantly longer than the FC method. These data strongly support the potential of FC technology for rapid and accurate identification of medically important yeasts. With the introduction of new antifungals, rapid, accurate identification of pathogenic yeasts is more important than ever for guiding antifungal chemotherapy.

  17. Comparison of T-SPOT.TB assay and tuberculin skin test for the evaluation of young children at high risk for tuberculosis in a community setting.

    Science.gov (United States)

    Nicol, Mark P; Davies, Mary-Ann; Wood, Kathryn; Hatherill, Mark; Workman, Lesley; Hawkridge, Anthony; Eley, Brian; Wilkinson, Katalin A; Wilkinson, Robert J; Hanekom, Willem A; Beatty, David; Hussey, Gregory

    2009-01-01

    We wished to compare the sensitivity of an enzyme-linked immunospot assay (T-SPOT.TB; Oxford Immunotec, Oxford, United Kingdom) and the tuberculin skin test for the detection of tuberculosis infection in very young children being evaluated for active tuberculosis in a rural community setting. Children with a history of exposure to tuberculosis and children presenting to a local clinic or hospital with symptoms suggesting tuberculosis were admitted to a dedicated case verification ward. T-SPOT.TB testing was performed, and children were evaluated with a clinical examination, a tuberculin skin test, chest radiographs, and cultures of induced sputum and gastric lavage specimens. The diagnosis was determined by using a clinical algorithm. A total of 243 children (median age: 18 months) were recruited, of whom 214 (88%) had interpretable T-SPOT.TB results. Children > or =12 months of age were more likely than younger children to have positive T-SPOT.TB results, whereas tuberculin skin test results were unaffected by age. The sensitivity of the T-SPOT.TB was no better than that of the tuberculin skin test for culture-confirmed tuberculosis (50% and 80%, respectively) and was poorer for the combined group of culture-confirmed and clinically probable tuberculosis (40% and 52%, respectively). For the 50 children clinically categorized as not having tuberculosis, the specificity of both the T-SPOT.TB and the tuberculin skin test was 84%. For young children presenting in a community setting after exposure to tuberculosis or with symptoms suggesting tuberculosis, T-SPOT.TB cannot be used to exclude active disease. The sensitivity of this assay may be impaired for very young children.

  18. Structural comparison of O-antigen gene clusters of Legionella pneumophila and its application of a serogroup-specific multiplex PCR assay.

    Science.gov (United States)

    Cao, Boyang; Tian, Zhenyang; Wang, Suwei; Zhu, Zhiyan; Sun, Yamin; Feng, Lu; Wang, Lei

    2015-12-01

    The Legionella pneumophila serogroups O1, O4, O6, O7, O10 and O13 are pathogenic strains associated with pneumonia. The surface O-antigen gene clusters of L. pneumophila serogroups O4, O6, O7, O10 and O13 were sequenced and analyzed, with the function annotated on the basis of homology to that of the genes of L. pneumophila serogroup O1 (L. pneumophila subsp. pneumophila str. Philadelphia 1). The gene locus of the six L. pneumophila serogroups contains genes of yvfE, neuABCD, pseA-like for nucleotide sugar biosynthesis, wecA for sugar transfer, and wzm as well as wzt for O-antigen processing. The detection of O-antigen genes allows the fine differentiation at species and serogroup level without the neccessity of nucleotide sequencing. The O-antigen-processing genes wzm and wzt, which were found to be distinctive for different for different serogroups, have been used as the target genes for the detection and identification of L. pneumophila strains of different O serogroups. In this report, a multiplex PCR assay based on wzm or wzt that diferentiates all the six serogroups by amplicon size was developed with the newly designed specific primer pairs for O1 and O7, and the specific primer pairs for O4, O6, O10, and O13 reported previously. The array was validated by analysis of 34 strains including 15 L. pneumophila O-standard reference strains, eight reference strains of other Legionella non-pneumophila species, six other bacterial species, and five L. pneumophila environmental isolates. The detection sensitivity was one ng genomic DNA. The accurate and sensitive assay is suitable for the identification and detection of strains of these serogroups in environmental and clinical samples.

  19. Comparison of different assays for detection of Chlamydia trachomatis%沙眼衣原体感染不同检测方法的比较

    Institute of Scientific and Technical Information of China (English)

    杜昆

    2014-01-01

    目的:研究不同方法检测沙眼衣原体感染的灵敏度和特异度。方法采取213例女性非淋菌性泌尿生殖道感染者宫颈分泌物,分别用直接免疫荧光法(DFA)、免疫层析法(ICA)和实时荧光定量PCR法(FQ-PCR)检测沙眼衣原体。结果采用DAF法和FQ-PCR法与采用ICA法检测沙眼衣原体阳性率比较差异有统计学意义(P<0.05);DFA法、ICA法和FQ-PCR法的灵敏度分别为95.1%、60.2%和97.3%,特异度分别为93.2%、99.2%和99.3%。DFA法和FQ-PCR法的灵敏度高于ICA法,差异有统计学意义(P<0.05),ICA法和FQ-PCR法的特异度高于DFA法,差异有统计学意义(P<0.05)。结论 FQ-PCR法有较高的灵敏度和特异度,可为临床诊断沙眼衣原体感染提供可靠依据。基层医疗卫生单位适合用IC A法检测沙眼衣原体。%Objective To investigate the sensitivity and specificity of different assays for detection of Chlamydia trachomatis . Methods Chlamydia trachomatis was determined in samples of cervical secretions from 213 patients with nongonococcal urethritis or genitourinary tract infection by direct immunofluorescence assay (DFA ) ,gold-immunochromatographic assay (ICA ) and real-time fluorescence quantitative polymerase chain reaction (FQ-PCR) .Results Comparing the positive rates of Chlamydia trachoma-tis detected by adopting the DAF ,FQ-PCR and the ICA methods showed the statistical difference (P<0 .05) .The sensitivity and specificity of Chlamydia trachomatis detected by DFA ,ICA and FQ-PCR were 95 .1% ,60 .2% ,97 .3% and 93 .2% ,99 .2% , 99 .3% ,respectively .The sensitivity of the DFA and FQ-PCR methods was higher than that of the ICA method ,difference was sta-tistically significant(P<0 .05) ,The specificity of the ICA and FQ-PCR methods was higher than that of the DFA method ,differ-ence was statistically significant (P<0 .05) .Conclusion The FQ-PCR method has higher sensitivity and

  20. Comparison between optical coherence tomography technique and mechanical compression assay to evaluate ionizing radiation effects in frozen and lyophilized bone Tissue

    Energy Technology Data Exchange (ETDEWEB)

    Santin, Stefany Plumeri; Freitas, Anderson Zanardi de; Martinho Junior, Antonio Carlos; Dias, Djalma Batista; Soares, Fernando Augusto Neves; Pino, Eddy Segura; Veloso, Marcelo Noronha; Mathor, Monica B., E-mail: spsantin@usp.br, E-mail: mathor@ipen.br [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil); Santos, Luiz Augusto Ubirajara, E-mail: augustosantos@terra.com.br [Universidade de Sao Paulo (IOT/HCFUSP), Sao Paulo, SP (Brazil). Fac. de Medicina. Instituto de Ortopedia e Traumatologia

    2013-07-01

    Currently tissue banks have utilized ionizing radiation to sterilize bone tissues to be used as allograft. This method is advantageous when compared with other techniques, because the tissue is sterilized in its final packaging avoiding later contaminations, another advantage is due to the fact occur only a minimal increase in temperature, in addition to provide a Sterility Assurance Level (SAL) of 10{sup -6}, as recommended by national and international standards. However, there are several studies investigating the modifications that this method of sterilization may cause to the bone matrix, for example, alterations in the resistance to compression force. The compressive mechanical tests are highly used to evaluate the decrease in the mechanical strength; however it is a destructive assay. In this study, we used Optical Coherence Tomography to evaluate these possible changes. This technique is advantageous, for do not destroy the sample and enable the performing of other assays with the same sample. In literature, it is possible to find several studies about mechanical changes occasioned by destructive tests. Therefore, this study aims to compare the results of both techniques. It was selected four donors to obtain eight samples of fibula, through a partnership with the Tissue Bank (Instituto de Traumatologia do Hospital das Clinicas da Universidade de Sao Paulo). From each donor were separated twelve samples for preservation by freezing and twelve samples for preservation by lyophilization. The samples were analyzed by Optical Coherence Tomography (OCT) after irradiation at different doses (15, 25 and 50 kGy), in addition to non-irradiated control. After the samples were analyzed by Optical Coherence Tomography the same were subjected to mechanical testing. The data were analyzed by software developed by Dr. Anderson Zanardi de Freitas to calculate the total attenuation coefficient of photons. Nevertheless, only the preservation method may induce to alterations

  1. Validation of a heterologous fertilization assay and comparison of fertilization rates of equine oocytes using in vitro fertilization, perivitelline, and intracytoplasmic sperm injections.

    Science.gov (United States)

    Sessions-Bresnahan, D R; Graham, J K; Carnevale, E M

    2014-07-15

    IVF in horses is rarely successful. One reason for this could be the failure of sperm to fully capacitate or exhibit hyperactive motility. We hypothesized that the zona pellucida (ZP) of equine oocytes prevents fertilization in vitro, and bypassing the ZP would increase fertilization rates. Limited availability of equine oocytes for research has necessitated the use of heterologous oocyte binding assays using bovine oocytes. We sought to validate an assay using bovine oocytes and equine sperm and then to demonstrate that bypassing the ZP using perivitelline sperm injections (PVIs) with equine sperm capacitated with dilauroyl phosphatidylcholine would result in higher fertilization rates than standard IVF in bovine and equine oocytes. In experiment 1, bovine oocytes were used for (1) IVF with bovine sperm, (2) IVF with equine sperm, and (3) intracytoplasmic sperm injections (ICSIs) with equine sperm. Presumptive zygotes were either stained with 4',6-diamidino-2-phenylindole from 18 to 26 hours at 2-hour intervals or evaluated for cleavage at 56 hours after addition of sperm. Equine sperm fertilized bovine oocytes; however, pronuclei formation was delayed compared with bovine sperm after IVF. The delayed pronuclear formation was not seen after ICSI. In experiment 2, bovine oocytes were assigned to the following five groups: (1) cumulus oocyte complexes (COCs) coincubated with bovine sperm; (2) COC exposed to sucrose then coincubated with bovine sperm; (3) COC coincubated with equine sperm; (4) COC exposed to sucrose, and coincubated with equine sperm; and (5) oocytes exposed to sucrose, and 10 to 15 equine sperm injected into the perivitelline (PV) space. Equine sperm tended (P = 0.08) to fertilize more bovine oocytes when injected into the PV space than after IVF. In experiment 3, oocytes were assigned to the following four groups: (1) IVF, equine, and bovine COC coincubated with equine sperm; (2) PVI of equine and bovine oocytes; (3) PVI with equine oocytes

  2. 6 HCV genotyping 9G test and its comparison with VERSANT HCV genotype 2.0 assay (LiPA) for the hepatitis C virus genotyping.

    Science.gov (United States)

    Chantratita, Wasun; Song, Keum-Soo; GunHo, Choi; Pongthanapisith, Viroj; Thongbaiphet, Nipa; Wongtabtim, Garanyuta; Pasomsub, Ekawat; Angkanavin, Kanokwan; Nimse, Satish Balasaheb; Sonawane, Mukesh Digambar; Warkad, Shrikant Dasharath; Kim, Taisun

    2017-01-01

    In this article, we describe the 6 HCV Genotyping 9G test and its evaluation by using clinical samples and plasmid DNA standards. In tests with 981 plasmid DNA standards, the 6 HCV Genotyping 9G test showed higher than 92.5% sensitivity and 99.4% specificity. The 6 HCV Genotyping 9G test was compared with the VERSANT HCV Genotype 2.0 assay (LiPA 2.0) for detection and discrimination of HCV genotypes in clinical samples. The results of both tests were verified by genomic sequencing. The 6 HCV Genotyping 9G test demonstrated a 100% agreement with the sequencing results, which was higher than LiPA 2.0. These results indicate that the 6 HCV Genotyping 9G test can be a reliable, sensitive, and accurate diagnostic tool for the correct identification of HCV genotypes in clinical specimens. 6 HCV Genotyping 9G test can genotype six HCV types in 1 PCR in 30min after PCR amplification. The 6 HCV Genotyping 9G test, thus provide critical information to physicians and assist them to apply accurate drug regimen for the effective hepatitis C treatment.

  3. Comparison of Enzyme-Linked Immunosorbent Assay, Surface Plasmon Resonance and Biolayer Interferometry for Screening of Deoxynivalenol in Wheat and Wheat Dust

    Directory of Open Access Journals (Sweden)

    Melanie Sanders

    2016-04-01

    Full Text Available A sample preparation method was developed for the screening of deoxynivalenol (DON in wheat and wheat dust. Extraction was carried out with water and was successful due to the polar character of DON. For detection, an enzyme-linked immunosorbent assay (ELISA was compared to the sensor-based techniques of surface plasmon resonance (SPR and biolayer interferometry (BLI in terms of sensitivity, affinity and matrix effect. The matrix effects from wheat and wheat dust using SPR were too high to further use this screenings method. The preferred ELISA and BLI methods were validated according to the criteria established in Commission Regulation 519/2014/EC and Commission Decision 2002/657/EC. A small survey was executed on 16 wheat lots and their corresponding dust samples using the validated ELISA method. A linear correlation (r = 0.889 was found for the DON concentration in dust versus the DON concentration in wheat (LOD wheat: 233 μg/kg, LOD wheat dust: 458 μg/kg.

  4. Comparison of post-thaw DNA integrity of boar spermatozoa assessed with the neutral comet assay and Sperm-Sus Halomax test kit.

    Science.gov (United States)

    Fraser, L; Parda, A; Filipowicz, K; Strzeżek, J

    2010-10-01

    In this study, we tested the hypothesis whether the neutral Comet assay (NCA) and the Sperm-Sus-Halomax (SSH) test kit could provide similar measurements of post-thaw DNA fragmentation of boar spermatozoa. Whole ejaculates or sperm-rich fractions of boar semen were frozen in an extender containing lactose, lipoprotein fractions isolated from ostrich egg yolk (LPFo), glycerol (lactose-LPFo-G) or in a standard boar semen extender (K3), without the addition of cryoprotective substances. In all boars, both the NCA and SSH test showed similar levels of post-thaw sperm DNA fragmentation in samples of the same ejaculates, regardless of the ejaculate collection procedure and extender. Yet, the levels of post-thaw sperm DNA damage, detected by the NCA and SSH test, were more accentuated in spermatozoa frozen in the absence of cryoprotective substances. Both the NCA and SSH detected variations among individual boars in terms of post-thaw sperm DNA fragmentation. Agreement between the measurements of the NCA and SSH was confirmed by scatter plots of differences, suggesting that the DNA integrity tests could detect the same sperm populations, which were susceptible to cryo-induced DNA damage. The findings of this study indicate that the NCA and the SSH test are effective in detecting similar levels of sperm DNA fragmentation and reinforce their importance in the assessment of frozen-thawed boar semen quality.

  5. Comparison between indirect enzyme-linked immunosorbent assays for Anaplasma marginale antibodies with recombinant major surface protein 5 and initial body antigens

    Directory of Open Access Journals (Sweden)

    Virgínia MG Silva

    2006-08-01

    Full Text Available Indirect enzyme-linked immunosorbent assays (ELISAs based on recombinant major surface protein 5 (rMSP5 and initial body (IB antigens from a Brazilian isolate of Anaplasma marginale were developed to detect antibodies against this rickettsia in cattle. Both tests showed the same sensitivity (98.2% and specificities (100% for rMSP5 and 93.8% for IB ELISA which did not differ statistically. No cross-reactions were detected with Babesia bigemina antibodies, but 5 (rMSP5 ELISA to 15% (IB ELISA of cross-reactions were detected with B. bovis antibodies. However, such difference was not statistically significant. Prevalences of seropositive crossbred beef cattle raised extensively in Miranda county, state of Mato Grosso do Sul, Brazil, were 78.1% by rMSP5 ELISA and 79.7% by IB ELISA. In the analysis of sera from dairy calves naturally-infected with A. marginale, the dynamics of antibody production was very similar between both tests, with maternal antibodies reaching the lowest levels at 15-30 days, followed by an increase in the mean optical densities in both ELISAs, suggesting the development of active immunity against A. marginale. Results showed that all calves were seropositive by one-year old, characterizing a situation of enzootic stability. The similar performances of the ELISAs suggest that both tests can be used in epidemiological surveys for detection of antibodies to A. marginale in cattle.

  6. Diagnostic accuracy of an IgM enzyme-linked immunosorbent assay and comparison with 2 polymerase chain reactions for early diagnosis of human leptospirosis.

    Science.gov (United States)

    Vanasco, N B; Jacob, P; Landolt, N; Chiani, Y; Schmeling, M F; Cudos, C; Tarabla, H; Lottersberger, J

    2016-04-01

    Enzyme-linked immunosorbent assay (ELISA) tests and polymerase chain reaction (PCR) may play a key role for early detection and treatment of human leptospirosis in developing countries. The aims of this study were to develop and validate an IgM ELISA under field conditions and to compare the diagnostic accuracy among IgG, IgM ELISAs, conventional PCR (cPCR), and real-time PCR (rtPCR) for early detection of human leptospirosis. Overall accuracy of IgM ELISA was sensitivity of 87.9%, specificity of 97.0%, and area under the curve of 0.940. When the 4 methods were compared, IgM ELISA showed the greatest diagnostic accuracy (J=0.6) followed by rtPCR (J=0.4), cPCR (J=0.2) and IgG ELISA (J=0.1). Our results support the use of IgM ELISA and rtPCR for early diagnosis of the disease. Moreover, due to their high specificity, they could be also useful to replace or supplement microscopic agglutination test as a confirmatory test, allowing more confirmations.

  7. Comparison of interferon-γ release assays, QuantiFERON TB-GIT and T-Spot.TB, in renal transplantation.

    Science.gov (United States)

    Ishikawa, Satoru; Igari, Hidetoshi; Akutsu, Naotake; Tsuyuzaki, Mizue; Aoyama, Hiromichi; Hasegawa, Masayuki; Otsuki, Kazunori; Maruyama, Michihiro; Saigo, Kenichi; Suzuki, Kiminori; Yamagishi, Fumio

    2017-07-01

    Renal transplant recipients are at increased risk of reactivating latent tuberculosis infection (LTBI) and progressing to active tuberculosis (TB). This study was conducted in National hospital for tuberculosis and renal transplantation (RT) in Japan. The purpose is to compare two interferon-γ release assays (IGRAs), QuantiFERON(®)-TB Gold in Tube (QFT) and T-SPOT(®).TB (TSPOT), in patients after renal transplantation for detecting latent TB infection (LTBI). Total 92 renal transplant recipients (median age 46 years, range 17-75) were prospectively enrolled, and QFT and TSPOT were concurrently examined. Total subjects were 92 patients (median age 46 years, range 17-75). The positive rate in QFT and TSPOT were 6.5% (95% confidence interval (CI) 3.0-13.5) and 2.2% (95% CI 1.0-7.6), respectively. There was a significant difference in IGRAs positivity (P TB during median follow-up of 994 days. Neither QFT nor TSPOT reaches estimated TB infection rate in Japan, especially elderly recipients aged 60 year-old or more. Therefore, both IGRAs might underestimate LTBI owing to immune suppressive therapy and aging. Physicians for renal transplantation need to understand the characteristics of both IGRAs and pay attention to the possibility of developing active TB even in patients of negative IGRAs results. Copyright © 2017 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.