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Sample records for sandwich elisa method

  1. Hyperglucagonaemia analysed by glucagon sandwich ELISA

    DEFF Research Database (Denmark)

    Albrechtsen, Nicolai Jacob Wewer; Hartmann, Bolette; Veedfald, Simon

    2014-01-01

    characterised. The specific determination of fully processed, intact glucagon requires a 'sandwich' assay employing a combination of antibodies directed against both N- and C-termini. We compared a novel assay for intact glucagon with a highly sensitive C-terminal RIA (hitherto considered specific) to determine...... the extent to which the hyperglucagonaemia measured in clinical samples was caused by authentic glucagon. METHODS: We examined the performance of three commercial glucagon 'sandwich' ELISAs. The ELISA with the best overall performance was selected to compare glucagon measurements in clinical samples...

  2. Validation and comparison of a sandwich ELISA, two competitive ELISAs and a real-time PCR method for the detection of lupine in food.

    Science.gov (United States)

    Ecker, Christina; Ertl, Anna; Pulverer, Walter; Nemes, Albert; Szekely, Pal; Petrasch, Angelika; Linsberger-Martin, Gertrud; Cichna-Markl, Margit

    2013-11-01

    Methods applied in food allergen analysis should be specific, sensitive and applicable to both raw and highly processed foods. The performance of the most commonly used methods, ELISA and real-time PCR, may, however, be influenced by food processing steps, e.g., heat treatment. The present study compares the applicability of four in-house developed methods, one sandwich ELISA, two competitive ELISAs and a real-time PCR method, for the detection of lupine in four different food matrices, comprising bread, biscuits, rice patties and noodles. In order to investigate the influence of food processing on the detectability, not only the heat treated model foods but also the corresponding doughs were analysed. The sandwich ELISA proved to be the most sensitive method. The LOD was found to be 10 ppm lupine, independent from the food matrix and independent if the dough or the heat treated food was analysed. In addition, the methods were applied to the analysis of commercial foodstuffs differing in their labelling. Copyright © 2013 Elsevier Ltd. All rights reserved.

  3. Evaluation and selection of indirect ELISA and sandwich ELISA kits for anti-HCV detection

    Directory of Open Access Journals (Sweden)

    LI Yongzhen

    2014-10-01

    Full Text Available ObjectiveTo compare the sensitivity and specificity between indirect enzyme-linked immunosorbent assay (ELISA and double-antibody sandwich ELISA kits produced in China and to select the best ELISA kit. MethodsSamples for evaluation included 60 serum plates and 40 serum samples positive or weakly positive for antibody to hepatitis C virus (anti-HCV which were confirmed by recombinant immunoblot assay. These samples were tested with a sandwich ELISA kit and three indirect ELISA kits, all of which were produced in China. Comparison between ELISA kits was made by paired chi-square test; comparison of false negative rate was made by R×C contingency table test. ResultsThe sensitivities of three indirect ELISA kits and a sandwich ELISA kit were 90.2%, 78.0%, 95.1%, and 97.6%, respectively, and the specificities were 78.1%, 72.6%, 94.1%, and 100%, respectively. The sandwich ELISA kit had a 4-8 times higher sensitivity than indirect ELISA kits. The R×C contingency table test revealed significant differences in false negative rate between ELISA kits and combinations of ELISA kits (χ2=29.898, P<0.05. ConclusionSandwich ELISA kit has higher sensitivity and specificity than indirect ELISA kits. Combined use of sandwich ELISA and indirect ELISA kits can significantly reduce the false negative rate and effectively prevent missed anti-HCV detection.

  4. Establishment of a Sandwich ELISA Method for Detection of Vascular Endothelial Growth Factor in Serum Samples of Hepatocellular Carcinoma Patients1

    Institute of Scientific and Technical Information of China (English)

    BING SHAN; JING-QUN HU; PING ZHAO; JUN HAN; XIAO-PING DONG; CHEN GAO; JIAN-MING CHEN; XIN-YU BI; BAO-YUN ZHANG; YAN GUO; CHEN-FANG DONG; RUN AN; QI SHI

    2008-01-01

    Objective To establish a sandwich ELISA method for detecting vascular endothelial growth factor(VEGF)in sera of population and the patients with hepatocellular carcinoma(HCC). Methods Full length and two truncated human VEGF cDNA sequences were amplified from a commercial plasmid pBLAST49-bVEGF by PCR and inserted into the prokaryotic-expression plasmid pET-32a or pGEX-2T. Various VEGF proteins were expressed and purified from E. coli in His-Trx or GST fusion forms.The specific VEGF antibodies were elicited in experimental rabbits and mice by immunization of the full length VEGF fusion protein His-Trx-VEGF1-165.After purification of antibodies with chromatograph of Protein G.a sandwich ELISA technique was established.Serum VEGF levels were evaluated in 229 adults and 291 HCC patients.Results SDS-PAGE displayed that the molecular weights of the expressed full length(His-Trx-VEGF1-165),N-terminal(His-Trx-VEGF1-100)and C-terminal(GST-VEGF100-165)human VEGF fusion proteins were about 38KD,31KD,and 33KD,respectively.Western blots confirmed that the prepared antisera were able to recognize both prokaryoticly and eukaryoticly expressed recombinant VEGF proteins.Assays of serially diluted His-Trx-VEGF1-100 by the established sandwich ELISA method showed that the linear range of the standard curve was 0.625-320 ng/mL.with the squared correlation coefficient R2=0.991.Screening of a serum panel containing 291 serum samples of HCC patients and 229 health adults rovealed that the average VEGF level in HCC patients was higher than that in healthy controls,with a statically significant difference. Conclusion The established sandwich ELISA reflects the level of serum VEGF and provide scientific basis for scrcening metastasis and recurrence of HCC using serum VEGF as an index.

  5. Detection of Penicillinase in Milk by Sandwich ELISA Based Polyclonal and Monoclonal Antibody.

    Science.gov (United States)

    Zhao, Yinli; Li, Guoxi

    2016-01-01

    A sandwich ELISA has been developed using polyclonal and monoclonal antibody for the determination of penicillinase in milk. For this purpose, specific polyclonal and monoclonal antibodies against penicillinase were generated and characterized. Using penicillinase standards prepared from 1-128 ng/mL, the method indicated that the detection limit of the sandwich ELISA, as measured in an ELISA plate reader, was as low as 0.86 ng/mL of penicillinase. For determine the accuracy, raw milk containing 2, 8, 32, and 64 ng/mL of penicillinase were tested by sandwich ELISA. Recoveries were from 93-97.5%, and the coefficient of variation [CV (%)] were from 5.55-8.38%. For interassay reproducibility, recoveries were from 89.5-95.1%, the coefficient of variation [CV (%)] were from 5.26-9.58%. This sandwich ELISA provides a useful screening method for quantitative detection of penicillinase in milk.

  6. A sandwich ELISA to detect VHSV and IPNV in turbot

    OpenAIRE

    Vázquez-Brañas, M. (Manuel); Coll-Morales, J. (Julio); Estepa, A

    1994-01-01

    Abstract: The recent demonstration that reared turbot (Scophthalmus maximus L) is a natural host for salmonid rhabdoviruses has made their rapid detection relevant to these fish species. A unique protocol to select and use non-competitive monoclonal antibodies (Mabs) for two high-sensitivity sandwich ELISAs has been developed to detect both infectious pancreatic necrosis virus (IPNV) and viral haemorrhagic septicaemia virus (VHSV) in turbot kidney extracts to assess the possibility of using t...

  7. An ultra-specific and sensitive sandwich ELISA for imatinib using two anti-imatinib antibodies.

    Science.gov (United States)

    Saita, Tetsuya; Yamamoto, Yuta; Hosoya, Kazuhisa; Yamamoto, Yutaro; Kimura, Sakiko; Narisawa, Yutaka; Shin, Masashi

    2017-05-29

    The development of an immunoassay for a low-molecular-weight drug first requires the identification of specific antibodies that do not cross-react with the drug's metabolites. If two antibodies can simultaneously recognize the entire structure of the drug, we can then utilize them to establish an ultra-specific sandwich ELISA, free from interference due to the metabolic products of the drug. This paper reports an ultra-specific and sensitive sandwich ELISA for determination of the tyrosine kinase inhibitor imatinib using two anti-imatinib antibodies. The anti-imatinib antibodies were obtained by two partial structures of imatinib as haptens (2-(5-amino-2-methylanilino)-4-(3-pyridyl)pyrimidine and 4-{(4-methyl-1-piperazinyl)-methyl}-benzoate). Under optimized conditions, this sandwich ELISA shows a linear detection range from 64 pg mL(-1) to 8 ng mL(-1), and a limit of detection of approximately 64 pg mL(-1) for 100-μL samples. The ELISA is specific to imatinib and while there was no cross-reactivity with the major metabolite N-desmethyl-imatinib, slight cross-reactivity was found with metabolite pyridine-N-oxide-imatinib. This assay demonstrated significantly lower cross-reactivity with metabolites than competitive ELISAs. Using this assay, drug levels were easily measured in rat blood after oral administration of imatinib via a single dose of 30 mg kg(-1) or 100 mg kg(-1). The levels in rat serum measured by this ELISA were comparable with those measured by HPLC, and there was a strong correlation between the values determined by the two methods (y = 0.983x + 0.081, R(2) = 0.948). Thus, we have successfully developed the first specific and sensitive sandwich ELISA for imatinib using two anti-imatinib antibodies. This sandwich ELISA will be a valuable tool for therapeutic drug monitoring and pharmacokinetic studies of imatinib. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Development of monoclonal antibody-based sandwich ELISA for detection of dextran.

    Science.gov (United States)

    Wang, Sheng-Yu; Li, Zhe; Wang, Xian-Jiang; Lv, Sha; Yang, Yun; Zeng, Lian-Qiang; Luo, Fang-Hong; Yan, Jiang-Hua; Liang, Da-Feng

    2014-10-01

    Dextran as anti-nutritional factor is usually a result of bacteria activity and has associated serial problems during the process stream in the sugar industry and in medical therapy. A sensitive method is expected to detect dextran quantitatively. Here we generated four monoclonal antibodies (MAbs) against dextran using dextran T40 conjugated with bovine serum albumin (BSA) as immunogen in our lab following hybridoma protocol. Through pairwise, an MAb named D24 was determined to be conjugated with horseradish peroxidase (HRP) and was used in the establishment of a sensitive sandwich enzyme-linked immunosorbent assay (ELISA) method for determination of dextran, in which MAb D9 was chosen as a capture antibody. The detection limit and working scope of the developed sandwich ELISA method were 3.9 ng/mL and 7.8-500 ng/mL with a correlation coefficient of 0.9909. In addition, the cross-reaction assay demonstrated that the method possessed high specificity with no significant cross-reaction with dextran-related substances, and the recovery rate ranged from 96.35 to 102.00%, with coefficient of variation ranging from 1.58 to 6.94%. These results indicated that we developed a detection system of MAb-based sandwich ELISA to measure dextran and this system should be a potential tool to determine dextran levels.

  9. Monoclonal antibody-based sandwich ELISA for the detection of staphylococcal enterotoxin A.

    Science.gov (United States)

    Kuang, Hua; Wang, Wenbing; Xu, Liguang; Ma, Wei; Liu, Liqiang; Wang, Libing; Xu, Chuanlai

    2013-04-19

    A sensitive and specific monoclonal antibody-based sandwich enzyme-linked immunosorbent assay (ELISA) was established and validated for the detection of staphylococcal enterotoxin A (SEA). After routine fusion and selection, 10 monoclonal antibodies showed high affinity for SEA. An optimal pair for sandwich ELISA was selected by pairwise interaction analysis. After optimization, the limit of detection (LOD) and linear dynamic range of the method were established, and were found to be 0.0282 ng/mL and 0.06-2 ng/mL, respectively. The recovery in pure milk ranged from 82.67% to 111.95% and the intra- and inter-assay coefficients of variation ranged from 3.16% to 6.05% and from 5.16% to 10.79%, respectively. Cross-reactivity with staphylococcal enterotoxin B (SEB), staphylococcal enterotoxin C (SEC), staphylococcal enterotoxin D (SED), and staphylococcal enterotoxin E (SEE) in this method were insignificant. These results indicate that the sandwich ELISA method developed in our study is effective for routine identification of SEA in food samples.

  10. Sandwich ELISA for quantitative detection of human collagen prolyl 4-hydroxylase

    Directory of Open Access Journals (Sweden)

    Myllyharju Johanna

    2010-06-01

    Full Text Available Abstract Background We describe a method for specific, quantitative and quick detection of human collagen prolyl 4-hydroxylase (C-P4H, the key enzyme for collagen prolyl-4 hydroxylation, in crude samples based on a sandwich ELISA principle. The method is relevant to active C-P4H level monitoring during recombinant C-P4H and collagen production in different expression systems. The assay proves to be specific for the active C-P4H α2β2 tetramer due to the use of antibodies against its both subunits. Thus in keeping with the method C-P4H is captured by coupled to an anti-α subunit antibody magnetic beads and an anti-β subunit antibody binds to the PDI/β subunit of the protein. Then the following holoenzyme detection is accomplished by a goat anti-rabbit IgG labeled with alkaline phosphatase which AP catalyzes the reaction of a substrate transformation with fluorescent signal generation. Results We applied an experimental design approach for the optimization of the antibody concentrations used in the sandwich ELISA. The assay sensitivity was 0.1 ng of C-P4H. The method was utilized for the analysis of C-P4H accumulation in crude cell extracts of E. coli overexpressing C-P4H. The sandwich ELISA signals obtained demonstrated a very good correlation with the detected protein activity levels measured with the standard radioactive assay. The developed assay was applied to optimize C-P4H production in E. coli Origami in a system where the C-P4H subunits expression acted under control by different promoters. The experiments performed in a shake flask fed-batch system (EnBase® verified earlier observations that cell density and oxygen supply are critical factors for the use of the inducer anhydrotetracycline and thus for the soluble C-P4H yield. Conclusions Here we show an example of sandwich ELISA usage for quantifying multimeric proteins. The method was developed for monitoring the amount of recombinant C-P4H tetramer in crude E. coli extracts. Due

  11. Development of Double Antibody Sandwich ELISA for Detection of Duck or Goose Flavivirus

    Institute of Scientific and Technical Information of China (English)

    NIU Hui-min; LI Xiang-rui; LI Yin; HUANG Xin-mei; HAN Kai-kai; LIU Yu-zhuo; ZHAO Dong-min; ZHANG Jing-feng; LIU Fei; LI Tong-tong; ZHOU Xiao-bo

    2013-01-01

    In order to establish double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) for detection of duck or goose flavivirus, polyclonal antibody against the flavivirus strain JS804 in geese and monoclonal antibody against the E protein of flavivirus strain JS804 in geese were used as the capture antibody and detection antibody, respectively. The optimal dilution of the capture antibody and detecting antibody capable of detecting the flavivirus strain JS804 in geese were 1:3 200 and 1:160 in the check-board titration, respectively. The reaction time of sample was 1 h, and the optimal working dilution of HRP-labeled goat-anti-mouse IgG was 1:10 000. The positive standard value was 0.247 (OD450 nm). The geese flavivirus could be detected at a minimal concentration of 1.875μg mL-1. The ELISA had no cross-reaction with Newcastle disease virus (NDV), Avian influenza virus (AIV), Infectious bronchitis virus (IBV), Infectious bursal disease virus (IBDV), Duck hepatitis virus (DHV), and Gosling plague virus (GPV). Twenty clinical samples were detected by the DAS-ELISA and RT-PCR respectively, with the agreement rate of 75%. The results revealed that the DAS-ELISA possessed favorable specificity and higher sensitivity, indicating a suitable method for rapid detection of the duck or goose flavivirus.

  12. Development of indirect sandwich ELISA for determination of excretory-secretory antigens of Fasciola hepatica

    Directory of Open Access Journals (Sweden)

    Libertad Alzamora-Gonzales

    2016-05-01

    Full Text Available Fasciolosis is a cosmopolitan parasitosis medical-veterinary importance caused by Fasciola hepatica, which affects sheep, goats and cattle; and it affects man accidentally causing an epidemic-endemic infection difficult to diagnose. The aim was to develop an indirect sandwich ELISA with 3 antibodies for detecting excretory-secretory antigens of Fasciola hepatica (ESFh. For the development of indirect sandwich ELISA were used, as capture antibody, mouse polyclonal antibodies anti ESFh and polyclonal antibodies rabbit anti-ESFh as detection antibody, at the concentrations of 10 and 5 µg/mL respectively. The conjugate used was mouse monoclonal anti- total immunoglobulins rabbit linked to peroxidase (1/1000. Were analized 31 sheep fecal samples, and the results were compared with those obtained by direct coproparasitological examination (DC and counterimmunoelectrophoresis (CIEP. The detection limit obtained for indirect sandwich ELISA was 100 ng/mL. The test had a 100% sensitivity, 96.6% specificity, positive and negative predictive values of 50% and 96.6% respectively, in relation to DC test. Comparing with CIEP the specificity obtained for indirect sandwich ELISA was 93.5% and a negative predictive value of 100%. We concluded that indirect sandwich ELISA designed is able to detect metabolic antigens in ovine feces samples and can be used for Fasciola hepatica diagnosis.

  13. Detection of potentially allergenic hazelnut (Corylus avellana) residues in food: a comparative study with DNA PCR-ELISA and protein sandwich-ELISA.

    Science.gov (United States)

    Holzhauser, Thomas; Stephan, Oliver; Vieths, Stefan

    2002-10-09

    Allergen detection is of increasing interest for food labeling purposes. A comparative study with a commercial hazelnut-specific PCR-ELISA and a sandwich-type ELISA detecting hazelnut protein was performed to investigate to what extent immunochemical and DNA-based techniques would correlate in the detection of trace amounts of potentially allergenic hazelnut residues. Both methods were highly sensitive and allowed the detection of even hazelnut in complex food matrixes. The protein-ELISA was highly specific for hazelnut. However, some foods could lead to false-positive results at the 10 ppm level. The PCR-ELISA did not show any cross-reactions with non-hazelnut foods, thus reducing the probability of having false positives at the trace level. Forty-one commercial food products with and without hazelnut components on their labels were analyzed for the presence of hazelnut. Of the 27 products in which hazelnut components were detected, two samples were not identified by the protein-ELISA, and only one sample, namely one white chocolate having hazelnut protein, was not detected by PCR-ELISA. The good correlation of the results of PCR-ELISA and protein-ELISA suggested that both PCR-based and immunochemical techniques are suitable for reliable detection of potentially allergenic hazelnut residues in foods at the trace level.

  14. Enhancing expression of the pseudorabies virus glycoprotein E in yeast and its application in an indirect sandwich ELISA.

    Science.gov (United States)

    Wu, C-Y; Wu, C-W; Liao, C-M; Chien, M-S; Huang, C

    2017-09-01

    The purpose of this study was to produce a recombinant pseudorabies virus (PRV) glycoprotein E (gE) protein with the correct antigenicity for use as a low-cost diagnostic antigen. The gene fragment encoding the amino-terminal immunodominant region of PRV gE (codons 31-270) (gEN31-270) was codon optimized and expressed constitutively and secreted using a Pichia pastoris expression system. Yeast-expressed gEN31-270 (ygEN31-270) was harvested from the culture supernatant, and ygEN31-270 was shown to exhibit N-linked glycosylation. An indirect sandwich enzyme-linked immunosorbent assay (ELISA) was developed using ygEN31-270 as a coating antigen, and the results showed that the assay had high sensitivity and specificity, as well as almost perfect concordance with a commercial gE ELISA kit. The immunodominant region (amino acids 31-270) of gE was expressed successfully in P. pastoris using a codon optimization strategy. ygEN31-270 was secreted and N-glycosylated. The ygEN31-270-based indirect sandwich ELISA showed high sensitivity and specificity to detect gE-specific antibodies in swine serum samples. The ygEN31-270-based indirect sandwich ELISA may provide an alternative method for developing a diagnostic kit with easy manipulation and low cost. © 2017 The Society for Applied Microbiology.

  15. Qualitative and quantitative detection of T7 bacteriophages using paper based sandwich ELISA.

    Science.gov (United States)

    Khan, Mohidus Samad; Pande, Tripti; van de Ven, Theo G M

    2015-08-01

    Viruses cause many infectious diseases and consequently epidemic health threats. Paper based diagnostics and filters can offer attractive options for detecting and deactivating pathogens. However, due to their infectious characteristics, virus detection using paper diagnostics is more challenging compared to the detection of bacteria, enzymes, DNA or antigens. The major objective of this study was to prepare reliable, degradable and low cost paper diagnostics to detect viruses, without using sophisticated optical or microfluidic analytical instruments. T7 bacteriophage was used as a model virus. A paper based sandwich ELISA technique was developed to detect and quantify the T7 phages in solution. The paper based sandwich ELISA detected T7 phage concentrations as low as 100 pfu/mL to as high as 10(9) pfu/mL. The compatibility of paper based sandwich ELISA with the conventional titre count was tested using T7 phage solutions of unknown concentrations. The paper based sandwich ELISA technique is faster and economical compared to the traditional detection techniques. Therefore, with proper calibration and right reagents, and by following the biosafety regulations, the paper based technique can be said to be compatible and economical to the sophisticated laboratory diagnostic techniques applied to detect pathogenic viruses and other microorganisms.

  16. Perbedaan Metode ELISA Sandwich A dan B dalam Deteksi Antigen Membran Toxoplasma gondii

    Directory of Open Access Journals (Sweden)

    ADI PRAYITNO

    2004-11-01

    Full Text Available Spreading of toxoplasmosis to fetus can by placenta, so it caused theabortion, born dead or congenital defect. To diagnosis this disease for fixed the acute infection must get the significant increasing of IgG by the soft fee. The objections of this study are to know the difference between ELISA Sandwich A and B in detecting of membrane antigen of Toxoplasma gondii (T. gondii in placenta tissue of pregnant women three-semester I and II with spontaneous abortion in Surakarta. One hundred serum and placenta tissue samples of pregnant women three-semester I and II with spontaneous abortion are got from dr. Muwardi Hospital. IgM anti Toxo from serum was examined by Toxo ISAGA Kit and IgG anti Toxo by Toxo Screen DA Kit. Detecting of membrane antigen of T. goodie from placenta tissue were done by ELISA Sandwich A and B. The result of this experiment showed that 33% were positive IBM and or Gig anti Toxo. Detection of membrane antigen toward 33 samples with positive Toxo (IgG positive was highly significant different between ELISA Sandwich A (3% positive toward ELISA Sandwich B (72.7% positive.

  17. A double-sandwich ELISA for identification of monoclonal antibodies suitable for sandwich immunoassays

    Science.gov (United States)

    The sandwich immunoassay (sIA) is an invaluable technique for concentrating, detecting, and quantifying target antigens. The two critical components required are a capture antibody and a detection antibody, each binding a different epitope on the target antigen. The specific antibodies incorporated...

  18. 检测百日咳毒素的双抗体夹心ELISA法的建立%Establishment of a double antibody sandwich ELISA method for determination of pertussis toxin

    Institute of Scientific and Technical Information of China (English)

    潘殊男; 肖詹蓉; 张霖阳; 史雨舟; 简志华; 王宇星; 李世慧; 张萍

    2012-01-01

    目的 建立白喉-破伤风-无细胞百日咳疫苗生产中百日咳毒素(pertussis toxin,PT)含量的测定方法.方法 免疫家兔制备高效价抗PT血清,纯化抗PT多克隆抗体并进行酶标记,建立双抗体夹心ELISA法并对其进行验证.结果 建立的方法在0~ 20 ng/ml PT测量区间呈现最佳线性,相关系数>0.99,且重复性好.检测百日咳丝状血凝素和黏着素,结果均为阴性,说明该法特异性良好.试验内10次、不同试验间3次测定16.0、8.0、4.0、3.0 ng/ml PT,变异系数为2.8%~9.7%,回收率为95.7%~106.0%,精密度和准确度验证均符合常规质量控制要求,定量限度为3.0 ng/ml.结论 该方法能有效检测百日咳杆菌大罐发酵过程中分泌的PT,为PT质量控制奠定了重要基础.%Objective To develop an ELISA method for quantitative determination of pertussis toxin (PT) during production of combined diphtheria,tetanus and acellular pertussis vaccine.Methods After chinchilla rabbits were immunized by PT,high titer anti-serum against PT was obtained and polyclonal antibody against PT was prepared.Then,a double antibody sandwich ELISA method was developed and verified.Results The best linearity ranged from 0 to 20 ng/ml of PT (correlation coefficient > 0.99).No cross reactions with filamentous hemagglutinin and pertactin were observed by the developed ELISA.The variation coefficients of intra-and inter-assays were 2.8%-9.7% and the recovery rates were 95.7%-106.0% when 16.0,8.0,4.0 and 3.0 ng/ml of standard PTs were determined.The quantitative limit was identified as 3.0 ng/ml.Conclusion Due to the excellent specificity,precision,and accuracy,this method can be applied to detecting PT in fermentation broth quantitatively.

  19. Development of a sandwich Dot-ELISA for detecting bovine viral diarrhea virus antigen with E2 recombinant protein

    Institute of Scientific and Technical Information of China (English)

    Yuelan ZHAO; Yuzhu ZUO; Lei ZHANG; Jinghui FAN; Hanchun YANG; Jianhua QIN

    2009-01-01

    The IgG antibodies of rabbit anti-E2 protein of the bovine viral diarrhea virus were prepared by a general method from high efficiency serum immunized by E2 recombinant protein antigen expressed in E. coli prokaryotic expression system and were labeled to make enzymelabeled antibody with the method of NaIO4. A sandwich Dot enzyme-linked immunosorbent assay (Dot-ELISA) for the detection of BVDV was developed. The optimal reaction conditions of Dot-ELISAwere determined. The results show that optimal coating antibody was 300 μg·mL-1, the working concentration of HRP-labeled antibody was 1:50. The optimal blocking reagent and time were 5% bovine serum and 45 rain. The minimum detection of the content of antigen reached 1.35μg·mL-1. Compared with the routine IDEXX ELISA test kit with the whole virus, its specificity, sensitivity and coincidence rate were 90.48%, 96.55% and 95.24%, respectively. Compared with the sandwich Dot-ELISA with the negative staining electron microscope and RT-PCR, the coincidence rates were 90.9% and 93.1%, respectively. In addition, Bovine viral diarrhea virus (BVDV) antigen of 178 samples collected from cow farms in the Hebei Province, China, were detected by the developed Dot-ELISA and the IDEXX BVDV antigen Test Kit simultaneously, BVDV antigen positive rate was 39.89%-41.01%. The result of detecting clinical samples demonstrated that the established method showed its specificity, sensitivity and repeatability, whereas the results were easily interpreted without an ELISA reader.

  20. A sandwich ELISA for measurement of the primary glucagon-like peptide-1 metabolite.

    Science.gov (United States)

    Wewer Albrechtsen, Nicolai J; Asmar, Ali; Jensen, Frederik; Törang, Signe; Simonsen, Lene; Kuhre, Rune E; Asmar, Meena; Veedfald, Simon; Plamboeck, Astrid; Knop, Filip K; Vilsbøll, Tina; Madsbad, Sten; Nauck, Michael A; Deacon, Carolyn F; Bülow, Jens; Holst, Jens J; Hartmann, Bolette

    2017-09-01

    Glucagon-like peptide-1 (GLP-1) is an incretin hormone secreted from the gastrointestinal tract. It is best known for its glucose-dependent insulinotropic effects. GLP-1 is secreted in its intact (active) form (7-36NH2) but is rapidly degraded by the dipeptidyl peptidase 4 (DPP-4) enzyme, converting >90% to the primary metabolite (9-36NH2) before reaching the targets via the circulation. Although originally thought to be inactive or antagonistic, GLP-1 9-36NH2 may have independent actions, and it is therefore relevant to be able to measure it. Because reliable assays were not available, we developed a sandwich ELISA recognizing both GLP-1 9-36NH2 and nonamidated GLP-1 9-37. The ELISA was validated using analytical assay validation guidelines and by comparing it to a subtraction-based method, hitherto employed for estimation of GLP-1 9-36NH2 Its accuracy was evaluated from measurements of plasma obtained during intravenous infusions (1.5 pmol × kg(-1) × min(-1)) of GLP-1 7-36NH2 in healthy subjects and patients with type 2 diabetes. Plasma levels of the endogenous GLP-1 metabolite increased during a meal challenge in patients with type 2 diabetes, and treatment with a DPP-4 inhibitor fully blocked its formation. Accurate measurements of the GLP-1 metabolite may contribute to understanding its physiology and role of GLP-1 in diabetes. Copyright © 2017 the American Physiological Society.

  1. Sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of lupine residues in foods.

    Science.gov (United States)

    Kaw, C H; Hefle, S L; Taylor, S L

    2008-10-01

    Lupine has been increasingly used in food applications due to its high nutritional value and excellent functional properties. However, lupine provokes allergic reactions in susceptible individuals. The presence of undeclared lupine residues in foods can pose a serious health risk to lupine-allergic individuals. Therefore, the objective of this research was to develop a sandwich-type ELISA for the detection of lupine residues in foods. Lupine flour derived from Lupinus albus was used to immunize 3 rabbits and a sheep. Pooled lupine-specific antibodies were partially purified from the sera by ammonium sulfate precipitation. A sandwich lupine ELISA with a limit of quantification (LOQ) of 1 ppm was developed by utilizing the rabbit antisera as the capture reagent and the sheep antiserum as the detector reagent. The binding of the antigen-antibody complex was visualized by the addition of commercial rabbit antisheep IgG antibody labeled with alkaline phosphatase with subsequent addition of p-nitrophenyl phosphate substrate to produce a colored product for quantification. Minor cross-reactivity was observed with soy (Glycine max) and black bean (Castanospermum australe). The performance of the lupine ELISA was evaluated in reference food standards (beef frankfurter and apple cinnamon muffin) and laboratory-prepared cooked frankfurters and corn muffins. The mean percent recovery for lupine spiked-frankfurters and corn muffins were 108.4%+/- 8.8% and 103.1%+/- 11.5%, respectively. The sandwich-type lupine ELISA developed in this study provides food manufacturers and regulatory agencies with an effective analytical tool to detect and quantify lupine residues in processed foods.

  2. Development of a sandwich ELISA for quantification of immunoglobulin G in mink blood

    DEFF Research Database (Denmark)

    Mathiesen, Ronja; Chriél, Mariann; Struve, T.;

    A major concern amongst the Danish mink farmers is the incidence of the syndrome pre-weaning diarrhea. The syndrome causes major management issues and decreases the welfare of the mink and increases mortality in the pre-weaning period. The etiology of the syndrome is considered multifactorial...... as a specific cause is not fully established or understood. Adding to an increased risk of developing pre-weaning diarrhea is the fact that the mink kits are born with very low levels of circulating immunoglobulins. Rapid achievement of high levels of immunoglobulins in the bloodstream is essential for the kits...... early immunity and thus their resistance against pathogenic agents found in the environment. This study describes a sandwich ELISA for quantification of the concentration of total immunoglobulin G in mink blood. The ELISA was validated with serum samples from females (n=8) and their kits (litters of 4...

  3. Development of a monoclonal sandwich ELISA for direct detection of bluetongue virus 8 in infected animals.

    Science.gov (United States)

    Ten Haaf, Andre; Kohl, Johannes; Pscherer, Sibylle; Hamann, Hans-Peter; Eskens, Hans Ulrich; Bastian, Max; Gattenlöhner, Stefan; Tur, Mehmet Kemal

    2017-05-01

    Bluetongue is an infectious viral disease which can cause mortality in affected ruminants, and tremendous economic damage via impacts upon fertility, milk production and the quality of wool. The disease is caused by bluetongue virus (BTV) which is transmitted by species of Culicoides biting midge. Rapid detection of BTV is required to contain disease outbreaks and reduce economic losses. The purpose of this study was to develop a monoclonal sandwich ELISA for direct detection of BTV in infected animals. Phage display technology was used to isolate BTV specific antibody fragments by applying the human scFv Tomlinson antibody libraries directly on purified BTV-8 particles. Three unique BTV-8 specific human antibody fragments were isolated which were able to detect purified BTV particles and also BTV in serum of an infected sheep. A combination of a human/mouse scFv-Fc chimeric fusion protein and a human Fab fragment in a sandwich ELISA format was able to detect BTV specifically with a limit of detection (LOD) of 10(4) infectious virus particles, as determined by tissue culture titration. This approach provided pilot data towards the development of a novel diagnostic test that might be used for direct detection of BTV-8 particles. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Immunodiagnosis of paramphistomosis using monoclonal antibody-based sandwich ELISA for detection of Paramphistomum gracile circulating 16 kDa antigen.

    Science.gov (United States)

    Anuracpreeda, Panat; Tepsupornkul, Kullanid; Chawengkirttikul, Runglawan

    2017-06-01

    In this study, we have produced a monoclonal antibody (MoAb) against 16 kDa antigen of Paramphistomum gracile (16 kDaAgPg), and developed an accurate sandwich enzyme-linked immunosorbent assay (sandwich ELISA) for the detection of circulating 16 kDaAg in the serum and fecal samples from cattle naturally infected with P. gracile. MoAb 1D10 was immobilized on a microtitre plate, and the antigen in the samples was captured and detected with biotinylated rabbit anti-16 kDaAgPg antibody. The lower detection limit of sandwich ELISA was 3·5 pg mL-1, and no cross-reaction with other parasite antigens was evaluated. The reliability of the assay was examined using the serum and fecal samples from cattle naturally infected with P. gracile, Fasciola gigantica, Moniezia benedeni, Trichuris sp., Strongyloides sp., strongylids and non-infected animals. The sandwich ELISA showed the sensitivity, specificity and accuracy at 98·33, 100 and 99·55% (serum samples), and 96·67, 100 and 99·09% (fecal samples). Therefore, this detection method is a rapid and excellent potential assay for the accurate diagnosis of paramphistomosis.

  5. Quantitative sandwich ELISA for the determination of lupine (Lupinus spp.) in foods.

    Science.gov (United States)

    Holden, Lise; Faeste, Christiane K; Egaas, Eliann

    2005-07-27

    The use of lupine in foods has increased considerably during the past decade, reflected by a corresponding increase in reported lupine-induced allergic incidents. Lupine allergy may arise either by primary sensitization or by clinical cross-reactivity in peanut-allergic persons. Detection of lupine proteins in food has previously been based on the use of patient serum. A novel sandwich enzyme-linked immunosorbent assay (ELISA) for the detection and quantification of lupine in processed foods was developed, using a polyclonal rabbit antilupine capture antibody and a biotinylated conjugate of the same antibody for detection. The antibody was highly specific for lupine, apart from minor cross-reactivities to other legumes. The assay had a detection limit of 1 mug/g and was successfully used to quantify lupine protein in various food matrixes. Recoveries ranged from 60 to 116%, while the intra-and interassay coefficients of variation were <6% and <21%, respectively.

  6. Quantitative sandwich ELISA for determination of traces of hazelnut (Corylus avellana) protein in complex food matrixes.

    Science.gov (United States)

    Holzhauser, T; Vieths, S

    1999-10-01

    A hazelnut-specific sandwich-type ELISA based on polyclonal antisera was developed for detection of hidden hazelnut protein residues in complex food matrixes. In the absence of a food matrix, extractable protein from different native and toasted hazelnuts was detected at rates of 94 +/- 13 and 96 +/- 7% applying standards prepared from native and toasted hazelnuts, respectively. From complex food matrixes, 0.001-10% of hazelnut was recovered between 67 and 132%, in average by 106 +/- 17%. Depending on the food matrix, hazelnut protein could be detected down to the ppb (ng/g) level. Intraassay precision was hazelnut >/= 0.001% and interassay precision was hazelnut >/= 0.01%. In 12 of 28 commercial food products without labeling or declaration of hazelnut components, between 2 and 421 ppm of hazelnut protein was detected, demonstrating a remarkable presence of potentially allergenic hazelnut protein "hidden" in commercial food products.

  7. A sandwich ELISA for the conformation-specific quantification of the activated form of human Bax.

    Science.gov (United States)

    Teijido, Oscar; Ganesan, Yogesh Tengarai; Llanos, Raul; Peton, Ashley; Urtecho, Jean-Baptiste; Soprani, Adauri; Villamayor, Aimee; Antonsson, Bruno; Manon, Stéphen; Dejean, Laurent

    2016-03-15

    Bcl-2 family proteins are critical regulators of mitochondrial outer membrane permeabilization (MOMP), which represents the point of no return of apoptotic cell death. The exposure of the Bax N-terminus at the mitochondria reflects Bax activation; and this activated configuration of the Bax protein is associated with MOMP. N-terminal exposure can be detected using specific monoclonal and/or polyclonal antibodies, and the onset of activated Bax has extensively been used as an early marker of apoptosis. The protocols of immunoprecipitation and/or immunocytochemistry commonly used to detect activated Bax are long and tedious, and allow semiquantification of the antigen at best. The sandwich ELISA protocol we developed has a 5 ng/mL detection limit and is highly specific for the activated conformation of Bax. This ELISA allows a rapid quantification of activated human Bax in whole cells and isolated mitochondria protein extracts. These properties grant this assay the potential to further clarify the prognostic and diagnostic value of activated Bax in disorders associated with deregulated apoptotic pathways such as degenerative diseases or cancer. Copyright © 2016 Elsevier Inc. All rights reserved.

  8. Development of a double-monoclonal antibody sandwich ELISA: Tool for chicken interferon-γ detection ex vivo

    Science.gov (United States)

    Dai, Hua; Xu, Zheng-zhong; Wang, Meiling; Chen, Jun-hua; Chen, Xiang; Pan, Zhi-ming; Jiao, Xin-an

    2016-01-01

    The aim of the present work was to develop reagents to set up a chicken interferon-γ (ChIFN-γ) assay. Four monoclonal antibodies (mAbs) specific for ChIFN-γ were generated to establish sandwich ELISA based on 2 different mAbs. To improve the detection sensitivity of ChIFN-γ, a double-monoclonal antibody sandwich ELISA was developed using mAb 3E5 as capture antibody and biotinylated mAb 3E3 as a detection reagent. The results revealed that this ELISA has high sensitivity, allowing for the detection of 125 to 500 pg/mL of recombinant ChIFN-γ, and also has an excellent capacity for detecting native ChIFN-γ. This ELISA was then used to detect ChIFN-γ level in chickens immunized with a Newcastle disease virus (NDV) vaccine, the immunized chicken splenocytes were stimulated by NDV F protein as recall antigen. From our results, it appears that the sensitivity range of this sandwich ELISA test is adequate to measure the ex vivo release of ChIFN-γ. PMID:27127340

  9. Preparation of anti-ciguatoxin monoclonal antibodies using synthetic haptens: sandwich ELISA detection of ciguatoxins.

    Science.gov (United States)

    Tsumuraya, Takeshi; Fujii, Ikuo; Hirama, Masahiro

    2014-01-01

    Ciguatera fish poisoning (CFP) is a form of food poisoning caused by the consumption of fish that have accumulated a type of sodium channel activator toxin called ciguatoxins (CTXs), which are produced by dinoflagellates of the genus Gambierdiscus through the food chain. CFP affects more than 50000 people each year. The extremely low level of CTXs in tainted fish has hampered the development of antibodies for the detection of these toxins. Monoclonal antibodies (mAbs) specific against major congeners of CTX3C, 51-hydroxyCTX3C, CTX1B, and 54-deoxyCTX1B were prepared by immunization of mice with protein conjugates of rationally designed synthetic haptens in place of the natural toxins. We found that haptenic groups possessing a surface area larger than 400 angstroms2 were required to produce mAbs that can bind strongly to CTXs. Direct sandwich ELISA utilizing two different monoclonal antibodies that bind specifically to one of the two wings of a CTX were established to detect CTXs. No cross-reactivity was observed against the other marine toxins tested, including brevetoxin A, brevetoxin B, okadaic acid, and maitotoxin.

  10. A High-Throughput, Precipitating Colorimetric Sandwich ELISA Microarray for Shiga Toxins

    Directory of Open Access Journals (Sweden)

    Andrew Gehring

    2014-06-01

    Full Text Available Shiga toxins 1 and 2 (Stx1 and Stx2 from Shiga toxin-producing E. coli (STEC bacteria were simultaneously detected with a newly developed, high-throughput antibody microarray platform. The proteinaceous toxins were immobilized and sandwiched between biorecognition elements (monoclonal antibodies and pooled horseradish peroxidase (HRP-conjugated monoclonal antibodies. Following the reaction of HRP with the precipitating chromogenic substrate (metal enhanced 3,3-diaminobenzidine tetrahydrochloride or DAB, the formation of a colored product was quantitatively measured with an inexpensive flatbed page scanner. The colorimetric ELISA microarray was demonstrated to detect Stx1 and Stx2 at levels as low as ~4.5 ng/mL within ~2 h of total assay time with a narrow linear dynamic range of ~1–2 orders of magnitude and saturation levels well above background. Stx1 and/or Stx2 produced by various strains of STEC were also detected following the treatment of cultured cells with mitomycin C (a toxin-inducing antibiotic and/or B-PER (a cell-disrupting, protein extraction reagent. Semi-quantitative detection of Shiga toxins was demonstrated to be sporadic among various STEC strains following incubation with mitomycin C; however, further reaction with B-PER generally resulted in the detection of or increased detection of Stx1, relative to Stx2, produced by STECs inoculated into either axenic broth culture or culture broth containing ground beef.

  11. Determination of Aspergillus pathogens in agricultural products by a specific nanobody-polyclonal antibody sandwich ELISA.

    Science.gov (United States)

    Wang, Ting; Li, Peiwu; Zhang, Qi; Zhang, Wen; Zhang, Zhaowei; Wang, Tong; He, Ting

    2017-06-28

    Aspergillus and its poisonous mycotoxins are distributed worldwide throughout the environment and are of particular interest in agriculture and food safety. In order to develop a specific method for rapid detection of Aspergillus flavus to forecast diseases and control aflatoxins, a nanobody, PO8-VHH, highly reactive to A. flavus was isolated from an immunized alpaca nanobody library by phage display. The nanobody was verified to bind to the components of extracellular and intracellular antigen from both A. flavus and A. parasiticus. To construct a sandwich format immunoassay, polyclonal antibodies against Aspergillus were raised with rabbits. Finally, a highly selective nanobody-polyclonal antibody sandwich enzyme-linked immunosorbent assay was optimized and developed. The results revealed that the detection limits of the two fungi were as low as 1 μg mL(-1), and that it is able to detect fungal concentrations below to 2 μg mg(-1) of peanut and maize grains in both artificially and naturally contaminated samples. Therefore, we here provided a rapid and simple method for monitoring Aspergillus spp. contamination in agricultural products.

  12. Diagnostic efficacy of monoclonal antibody based sandwich enzyme linked immunosorbent assay (ELISA for detection of Fasciola gigantica excretory/secretory antigens in both serum and stool

    Directory of Open Access Journals (Sweden)

    Zoheiry Mona K

    2011-09-01

    Full Text Available Abstract Background This research was carried out to develop a reliable monoclonal antibody (MoAb-based sandwich enzyme linked immunosorbent assay (ELISA for the diagnosis of active Fasciola gigantica infection in both serum and stool for comparative purposes. Methods From a panel of MoAbs raised against F. gigantica excretory/secretory antigens (ES Ags, a pair (12B/11D/3F and 10A/9D/10G was chosen due to its high reactivity and strict specificity to F. gigantica antigen by indirect ELISA. Results The two MoAbs were of the IgG1 and IgG2a subclasses, respectively. Using SDS-PAGE and EITB, the selected MoAbs recognized 83, 64, 45 and 26 kDa bands of ES Ags. The lower detection limit of ELISA assay was 3 ng/ml. In stool, the sensitivity, specificity and diagnostic efficacy of ELISA was 96%, 98.2 and 97.1%; while in serum they were 94%, 94.6% and 94.3%, respectively. Moreover, a positive correlation was found between ova count in stool of F. gigantica infected patients and the OD readings of ELISA in both stool and serum samples (r = 0.730, p Conclusions These data showed that the use of MoAb-based sandwich ELISA for the detection of F. gigantica coproantigens in stool specimens was superior to serum samples; it provides a highly efficient, non-invasive technique for the diagnosis of active F. gigantica infection.

  13. Implementation of microfluidic sandwich ELISA for superior detection of plant pathogens.

    Directory of Open Access Journals (Sweden)

    Numrin Thaitrong

    Full Text Available Rapid and economical screening of plant pathogens is a high-priority need in the seed industry. Crop quality control and disease surveillance demand early and accurate detection in addition to robustness, scalability, and cost efficiency typically required for selective breeding and certification programs. Compared to conventional bench-top detection techniques routinely employed, a microfluidic-based approach offers unique benefits to address these needs simultaneously. To our knowledge, this work reports the first attempt to perform microfluidic sandwich ELISA for Acidovorax citrulli (Ac, watermelon silver mottle virus (WSMoV, and melon yellow spot virus (MYSV screening. The immunoassay occurs on the surface of a reaction chamber represented by a microfluidic channel. The capillary force within the microchannel draws a reagent into the reaction chamber as well as facilitates assay incubation. Because the underlying pad automatically absorbs excess fluid, the only operation required is sequential loading of buffers/reagents. Buffer selection, antibody concentrations, and sample loading scheme were optimized for each pathogen. Assay optimization reveals that the 20-folds lower sample volume demanded by the microchannel structure outweighs the 2- to 4-folds higher antibody concentrations required, resulting in overall 5-10 folds of reagent savings. In addition to cutting the assay time by more than 50%, the new platform offers 65% cost savings from less reagent consumption and labor cost. Our study also shows 12.5-, 2-, and 4-fold improvement in assay sensitivity for Ac, WSMoV, and MYSV, respectively. Practical feasibility is demonstrated using 19 real plant samples. Given a standard 96-well plate format, the developed assay is compatible with commercial fluorescent plate readers and readily amendable to robotic liquid handling systems for completely hand-free assay automation.

  14. Quantitation of Leishmania infantum in tissues of infected BALB/c mouse by sandwich ELISA.

    Science.gov (United States)

    Ferrua, B; Le Fichoux, Y; Suffia, I; Rousseau, D; Roptin, C; Kubar, J

    2001-01-01

    In this report, a sandwhich ELISA was developed to quantify spleen and liver burdens from L. infantum-infected BALB/c mice. Amastigote antigens obtained following Nonidet P40 extraction of parasite-harbouring tissues were captured by anti-L. infantum human IgG insolubilized onto microtiter plate and subsequently revealed with anti-L. infantum F(ab)' fragments labelled with peroxidase. The method was easy to perform, precise and capable to specifically and accurately detect 5 x 10(4) amastigotes/100 mg tissue. Parasite burdens from infected BALB/c mice, in various conditions, were measured by ELISA and Giemsa-stained touch imprint reference methods, and compared. Both techniques agreed well with close values for liver burdens, but the spleen loads measured by the ELISA were, on average, 10.7 times higher than those calculated from imprints. This difference was attributed partly to the underestimation brought by Stauber's formula. However, it did not preclude the usefulness of this newly developed test, since results obtained in kinetics studies and evaluation of the efficiency of leishmanicidal drugs allowed us to draw identical conclusions.

  15. Development of sandwich dot-ELISA for specific detection of Ochratoxin A and its application on to contaminated cereal grains originating from India

    Directory of Open Access Journals (Sweden)

    M. eVenkataramana

    2015-05-01

    Full Text Available In the present study, generation and characterization of a highly specific monoclonal antibody (mAb against Ochratoxin A (OTA was undertaken. The generated mAb was further used to develop a simple, fast and sensitive sandwich dot-ELISA (s-dot ELISA method for detection of OTA from contaminated food grain samples. The LOD (limit of detetion of the developed ELISA method was determined as 5.0 ng/mL of OTA. Developed method was more specific towards OTA and no cross reactivity was observed with the other tested mycotoxins such as deoxynivalenol, fumonisin B1 or aflatoxin B1. To assess the utility and reliability of the developed method, several field samples of maize, wheat and rice (n=195 collected from different geographical regions of southern Karnataka region of India were evaluated for the OTA occurrence. Seventy two out of 195 samples (19 maize, 38 wheat and 15 rice were found to be contaminated by OTA by s-dot ELISA. The assay results were further co-evaluated with conventional analytical HPLC method. Results of the s-dot ELISA are in concordance with HPLC except for 3 samples that were negative for OTA presence by s-dot ELISA but found positive by HPLC. Although positive by HPLC, the amount of OTA in the three samples was found to be lesser than the accepted levels (>5 µg/kg of OTA presence in cereals. Therefore, in conclusion, the developed s-dot ELISA is a better alternative for routine cereal based food and feed analysis in diagnostic labs to check the presence of OTA over existing conventional culture based, tedious analytical methods.

  16. Development of a sandwich ELISA-type system for the detection and quantification of hazelnut in model chocolates.

    Science.gov (United States)

    Costa, Joana; Ansari, Parisa; Mafra, Isabel; Oliveira, M Beatriz P P; Baumgartner, Sabine

    2015-04-15

    Hazelnut is one of the most appreciated nuts being virtually found in a wide range of processed foods. The simple presence of trace amounts of hazelnut in foods can represent a potential risk for eliciting allergic reactions in sensitised individuals. The correct labelling of processed foods is mandatory to avoid adverse reactions. Therefore, adequate methodology evaluating the presence of offending foods is of great importance. Thus, the aim of this study was to develop a highly specific and sensitive sandwich enzyme-linked immunosorbent assay (ELISA) for the detection and quantification of hazelnut in complex food matrices. Using in-house produced antibodies, an ELISA system was developed capable to detect hazelnut down to 1 mg kg(-1) and quantify this nut down to 50 mg kg(-1) in chocolates spiked with known amounts of hazelnut. These results highlight and reinforce the value of ELISA as rapid and reliable tool for the detection of allergens in foods.

  17. Detection of ruminant meat and bone meal in feeds by sandwich ELISA with monoclonal antibodies.

    Science.gov (United States)

    Yamamoto, Takayuki; Kato, Masatoshi; Endo, Kiwamu; Kotoura, Satoshi; Takeda, Zenya

    2016-01-01

    A sensitive and reproducible enzyme-linked immunosorbent assay (ELISA) using two monoclonal antibodies directed against a synthetic peptide with an amino-acid sequence related to the C-terminus of bovine myoglobin and the whole molecule of sodium dodecyl sulphate (SDS)-denatured bovine myoglobin was adapted for detecting bovine myoglobin in contaminated feeds. The ELISA employed bovine meat extract of a known myoglobin concentration as a calibration standard and had an limit of detection (LOD) of 3.54 ng/ml and an limit of quantification (LOQ) of 11.0 ng/ml corresponding to 0.022% and 0.067% (wt/wt) bovine meat-and-bone-meal (MBM) mixed in 20-fold-diluted feed extracts, respectively. A cut-off threshold of 20.6 ng/ml bovine myoglobin was set to simplify ELISA and facilitate quick assessment of test results without a tedious calibration process. The ELISA was able to detect bovine MBM in artificially prepared model feeds, mixed botanical feeds, mixed botanical feeds with skimmed milk, fish meal, pork meal and pork/chicken meal at 0.1% (wt/wt). It was also able to detect sheep MBM in test feeds, but showed no reactivity to swine MBM, chicken MBM, skimmed milk or gelatine of bovine origin. The advantages of this method are the quick and easy extraction protocol of proteins from test feeds, using 100 mM sodium sulphide and 0.6% sodium dodecyl sulphate in the extraction solution and the effective detection of bovine and sheep MBM at 0.1% (wt/wt).

  18. Development of a H2 O2 -sensitive quantum dots-based fluorescent sandwich ELISA for sensitive detection of bovine β-lactoglobulin by monoclonal antibody.

    Science.gov (United States)

    He, Shengfa; Li, Xin; Gao, Jinyan; Tong, Ping; Chen, Hongbing

    2017-06-16

    Bovine β-lactoglobulin (BLG) is the major allergen in cows' milk, and the specific epitope plays a key role in food allergy. Developing a method specifically bind to the IgE epitope is necessary for testing BLG and its allergenic residues. The monoclonal antibody (1G9) specific to the IgE linear epitope for BLG was identified as high affinity and specificity. Based on 1G9, a sensitive fluorescent sandwich enzyme-linked immunosorbent assay (sELISA) was successfully developed using catalase-mediated fluorescence quenching of thiolated CdTe quantum dots in the presence of hydrogen peroxide as fluorescent signal output. The fluorescent sELISA showed high sensitivity and specificity, the limit of detection was 0.49 ng mL(-1) , which was 16-fold lower than horseradish peroxidase (HRP)-based sELISA. The linear range for BLG detection were 125-4000 ng mL(-1) (r = 0.9939) and 0.48-62.5 ng mL(-1) (r = 0.9919). The recoveries and coefficients of variation were 94.25-109.83% and 4.38-20.29%, respectively. Allergenic residues were also detected in hydrolysed infant formulas. The results of fluorescent sELISA showed good performance as HRP-based sELISA and commercial sELISA kit. This proposed fluorescent sELISA could be employed to detect BLG and its allergenic residues in food with highly sensitivity, reliability, and recovery. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  19. Direct ELISA.

    Science.gov (United States)

    Lin, Alice V

    2015-01-01

    First described by Engvall and Perlmann, the enzyme-linked immunosorbent assay (ELISA) is a rapid and sensitive method for detection and quantitation of an antigen using an enzyme-labeled antibody. Besides routine laboratory usage, ELISA has been utilized in medical field and food industry as diagnostic and quality control tools. Traditionally performed in 96-well or 384-well polystyrene plates, the technology has expanded to other platforms with increase in automation. Depending on the antigen epitope and availability of specific antibody, there are variations in ELISA setup. The four basic formats are direct, indirect, sandwich, and competitive ELISAs. Direct ELISA is the simplest format requiring an antigen and an enzyme-conjugated antibody specific to the antigen. This chapter describes the individual steps for detection of a plate-bound antigen using a horseradish peroxidase (HRP)-conjugated antibody and luminol-based enhanced chemiluminescence (ECL) substrate. The methodological approach to optimize the assay by chessboard titration is also provided.

  20. Development and evaluation of a sandwich ELISA for quantification of the 20S proteasome in human plasma

    DEFF Research Database (Denmark)

    Dutaud, Dominique; Aubry, Laurent; Henry, Laurent

    2002-01-01

    Because quantification of the 20S proteasome by functional activity measurements is difficult and inaccurate, we have developed an indirect sandwich enzyme-linked immunosorbent assays (ELISA) for quantification of the 20S proteasome in human plasma. This sandwich ELISA uses a combination...... of a monoclonal antibody (mcp 20) recognizing the C2-beta subunit of human 20S proteasome (Mr˜30,000) and a polyclonal rabbit anti-20S antibody which labels different subunits of the complex. The detection limit of the assay was established as 10 ng/ml (n=10, mean of zero standard+2 S.D.) and the recovery rate...... ranged from 96% to 104%. The within-run and between-run coefficients of variation (CV) ranges were 2.8–3.3 and 3.0–3.4, respectively. Using serial dilutions of plasma to which various amounts of purified 20S proteasome were added, a linear dose–response was observed between 102 and 2050 ng...

  1. A high-throughput, precipitating colorimetric sandwich ELISA microarray for shiga toxins

    Science.gov (United States)

    Shiga toxins 1 and 2 (Stx1 and Stx2) from Shiga toxin-producing E. coli (STEC) bacteria were simultaneously detected with a newly developed, high-throughput antibody microarray platform. The proteinaceous toxins were immobilized and sandwiched between biorecognition elements (monoclonal antibodies)...

  2. Establishment of a convenient sandwich-ELISA for direct quantification of glucosyltransferase-I: application for dual diagnosis of dental caries.

    Science.gov (United States)

    Hashizume-Takizawa, Tomomi; Shinozaki-Kuwahara, Noriko; Tomita, Naoya; Kurita-Ochiai, Tomoko

    2014-04-01

    Previously, we established a convenient enzyme-linked immunosorbent assay (ELISA) system targeting glucosyltransferase (GTF)-B derived from Streptococcus mutans for diagnosing caries risk. However, it has been reported that S. sobrinus possesses high cariogenicity and is more frequently detected in highly caries-susceptible patients than S. mutans is. S. sobrinus can secrete GTF-I, an important cariogenic factor for dental plaque formation, as well as S. mutans GTF-B. Therefore, in this study, we developed another feasible ELISA system targeting S. sobrinus GTF-I that would ensure caries risk determination by combined GTF-I and GTF-B levels. A readily measurable sandwich-ELISA system was devised, which consisted of monoclonal and polyclonal antibodies against GTF-I. The developed sandwich-ELISA system quantified the purified GTF-I with sensitivity and specificity, and a positive correlation was observed between the amount of GTF-I extracted from clinical plaque samples and S. sobrinus levels. Furthermore, high levels of GTF-I and GTF-B were detected using the sandwich-ELISA system in caries-susceptible subjects. These results indicate that the sandwich-ELISA system against GTF-I developed in this study is useful, and that the dual detection of the caries risk factors GTF-I and GTF-B is helpful for predicting caries risk.

  3. Standard Test Method for Sandwich Corrosion Test

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    2009-01-01

    1.1 This test method defines the procedure for evaluating the corrosivity of aircraft maintenance chemicals, when present between faying surfaces (sandwich) of aluminum alloys commonly used for aircraft structures. This test method is intended to be used in the qualification and approval of compounds employed in aircraft maintenance operations. 1.2 The values stated in SI units are to be regarded as the standard. The values given in parentheses are for information. 1.3 This standard may involve hazardous materials, operations, and equipment. This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use. Specific hazard statements appear in Section 9.

  4. A sandwich ELISA for the detection of neuraminidase of avian influenza A(H7N9) virus.

    Science.gov (United States)

    Yu, Yueyang; Zhang, Xi; Zhao, Baihui; Sun, Ying; Zhang, Xiaoguang; Bai, Tian; Lu, Jian; Li, Zi; Liu, Liqi; Wang, Dayan; Shu, Yuelong; Zhou, Jianfang; Qin, Kun

    2017-09-01

    Although exhibiting no or low virulence in poultry, avian influenza virus H7N9 has caused around 1400 confirmed human infections in China with a case-fatality rate of 30% since 2013. A highly pathogenic H7N9 virus (HP-H7), with the HA antigenicity distinct from the previous, were recently detected in patients and poultry. Therefore, convenient rapid diagnosis with reliability will allow early antiviral use and management for H7N9 infection. Here, a sandwich ELISA targeting the conserved viral antigen, neuraminidase (NA) was developed. The immunoassay employed mouse monoclonal antibody (mAb) 3C1 to specifically capture the N9 and 3E9 for the detection. Its limit of detection is 6.25ng/ml for N9 protein of A/Anhui/1/2013(H7N9, AH1/2013) and 0.125HAU/50μL for live virus, AH1/2013 and A/Environment/Jiangxi/28/2009 (H11N9), respectively. When applied to test the five clinic throat swabs from H7N9 patients confirmed by nuclear acid testing (NAT) using quantitative reverse-transcriptase polymerase chain reaction (Q-PCR), two samples showed positive result in sandwich ELISA while all were negative using commercial Flu A and H7 subtype rapid antigen tests (RAT). The ELISA using anti-N9 mAbs provided a valuable approach to detect H7N9 virus and quantify the N9 protein. Copyright © 2017. Published by Elsevier B.V.

  5. 抗CPV-2a单克隆抗体的制备及双抗体夹心ELISA检测方法的建立%Preparation of Monoclonal Antibody Against CPV-2a and Establishment of Double Antibody Sandwich ELISA Detection Method to CPV-2a

    Institute of Scientific and Technical Information of China (English)

    王净; 李刚; 王鹏; 曾妮; 穆秀明; 高志花; 王慧文; 史利军

    2011-01-01

    The canine parvovirus (CPV) , which is hard to get cured, is one of the most severe diseases in dogs.However, an effective therapeutic method for the CPV is the monoclonal antibody. This article described the approach to the preparation of anti-CPV-2a monoclonal antibody. The canine parvovirus antibodies were prepared from New Zealand rabbit and Balb/c mouse with purified CPV-2a antigen. After subcloned, 4 strains of positive hybridoma cells were obtained, which were named 1H9, 2B5, 2B7 and 2C7 respectively. Their immunoreactivity were identified by Westem blotting and their specificity were done by indirect ELISA. In order to detect the canine parvovirus, a double antibody sandwich ELISA method was established. The assay used rabbit anti-CPV-2a polyclonal antibody as the capture antibody, monoclonal antibody as the trace antibody and goat anti-mouse IgG HRP conjugation as the detection system. The optimal dilution of the capture antibody and the trace antibody capable of detecting the CPV-2a antigens was found to be 1: 800 and 1: 2 000 respectively in the check-board titration, and that of the detection system was 1: 4 000. The result showed that the immunoreaction of the monoclonal antibody with pET-32a-VP2 protein was positive and its reaction with RV or CDV antigen was negative. The detective sensitivity ofsandwich ELISA was 4.375 μg/mL. Compared with RB of USA ELISA kit, the coincidence rate was 95%.The preparation of monoclonal antibody would provide the basis to treat the canine parvovirus. The establishment of double antibody sandwich ELISA method would be a simple, quick and reliable method for screening large numbers of fecal samples of dogs suspected of CPV infection.%犬细小病毒病是危害养犬业的重要传染病之一,患病犬难以治愈.单克隆抗体治疗此病效果明显,本文介绍了制备抗CPV-2a单克隆抗体的方法.用纯化的犬细小病毒(canine parvovirus,CPV) 2a型分离株免疫新西兰大

  6. One-step antibody immobilization-based rapid and highly-sensitive sandwich ELISA procedure for potential in vitro diagnostics.

    Science.gov (United States)

    Vashist, Sandeep Kumar; Marion Schneider, E; Lam, Edmond; Hrapovic, Sabahudin; Luong, John H T

    2014-03-18

    An improved enzyme-linked immunosorbent (ELISA) assay using one-step antibody immobilization has been developed for the detection of human fetuin A (HFA), a specific biomarker for atherosclerosis and hepatocellular carcinoma. The anti-HFA formed a stable complex with 3-aminopropyltriethoxysilane (APTES) by ionic and hydrophobic interactions. The complex adsorbed on microtiter plates exhibited a detection range of 4.9 pg mL(-1) to 20 ng mL(-1) HFA, with a limit of detection of 7 pg mL(-1). Furthermore, an analytical sensitivity of 10 pg mL(-1) was achieved, representing a 51-fold increase in sensitivity over the commercial sandwich ELISA kit. The results obtained for HFA spiked in diluted human whole blood and plasma showed the same precision as the commercial kit. When stored at 4°C in 0.1 M phosphate-buffered saline (PBS, pH 7.4), the anti-HFA bound microtiter plates displayed no significant decrease in their functional activity after two months. The new ELISA procedure was extended for the detection of C-reactive protein, human albumin and human lipocalin-2 with excellent analytical performance.

  7. Development and validation of a sandwich ELISA for the determination of potentially allergenic sesame (Sesamum indicum) in food.

    Science.gov (United States)

    Redl, Gerda; Husain, Fatima T; Bretbacher, Ines E; Nemes, Albert; Cichna-Markl, Margit

    2010-10-01

    This paper presents a sandwich enzyme-linked immunosorbent assay (ELISA) that allows the determination of traces of sesame in food. Chicken anti-sesame antibodies, used as coating antibodies, and rabbit anti-sesame antibodies, used as secondary antibodies, were prepared by immunization with a protein extract of white, peeled sesame. The ELISA did not show any cross-reactivity with 19 food ingredients commonly found in sesame-containing foodstuffs such as seeds, nuts, and cereals. In whole grain bread, crisp toast, and snacks, the limit of detection (S/N = 3) was 0.5, 0.5, and 0.3 μg sesame protein/g, and the limit of quantification (S/N = 10) was 0.6, 0.8, and 1.4 μg sesame protein/g, respectively. The analysis of blank food matrices (whole grain bread, white bread, crisp toast, and snacks) spiked with sesame protein at four spike levels generally resulted in mean recoveries from 72% to 145%. In the case of spiking blank food matrices with sesame seeds, the ELISA proved to be more accurate for whole wheat cookies than for whole wheat bread.

  8. Development of sandwich ELISA for testing bovine β-lactoglobulin allergenic residues by specific polyclonal antibody against human IgE binding epitopes.

    Science.gov (United States)

    He, Shengfa; Li, Xin; Gao, Jinyan; Tong, Ping; Chen, Hongbing

    2017-07-15

    Bovine β-lactoglobulin (BLG) is the main allergen in cows' milk, and the most commonly used method for detecting BLG is enzyme-linked immunosorbent assay (ELISA). However, antibodies used in commercial ELISA kits do not recognize specifically BLG IgE epitopes. Here, an antibody specific to IgE linear epitopes for BLG was used to develop a sandwich ELISA using a rabbit anti-BLG polyclonal antibody. The linear range for BLG detection was 31.25-8000ng/mL and limit of detection was 1.96ng/mL. BLG content in dairy samples was determined, and there was a good agreement between this immunoassay and reversed-phase high-performance liquid chromatography with high recovery. Additionally, BLG content in food samples had an average recovery of 104.25%. Allergenic residues were also detected in hydrolyzed infant formulas. The method developed could be a practical approach to determine BLG and its allergenic residues in food with a high degree of sensitivity, reliability and recovery. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. Sandwich enzyme-linked immunosorbent assay (ELISA) for detection of cashew nut in foods.

    Science.gov (United States)

    Gaskin, Ferdelie E; Taylor, Steve L

    2011-01-01

    The presence of undeclared cashew can pose a health risk to cashew-allergic consumers. The food industry has the responsibility to declare the presence of cashews on packaged foods even when trace residues are or might be present. The objective of this study was to develop a rapid, sensitive, and specific enzyme-linked immunosorbent assay (ELISA) for the detection of cashew residues. Raw and roasted cashews were defatted and used separately to immunize sheep, goats, and rabbits. The cashew ELISA was developed using sheep and rabbit polyclonal anti-roasted cashew sera as capture and detector reagents, respectively, with visualization through an alkaline phosphatase-mediated substrate reaction. The cashew ELISA was shown to have a limit of quantification of 1 ppm (1 μg cashew/g). The ELISA was highly specific except that substantial cross-reactivity was noted with pistachio and a lesser degree of cross-reactivity was noted with hazelnut. The performance of the ELISA was assessed by manufacturing cookies, ice cream, and milk chocolate with added known amounts (0 to 1000 ppm) of cashew. The mean percent recoveries for ice cream, cookies, and milk chocolate were 118%± 2.9%, 84.3%± 4.0%, and 104%± 3.0%, respectively. In a limited retail survey, 4/5 retail samples with cashew declared on ingredient labels tested positive for cashew compared to 5/36 samples of foods with precautionary labels indicating the possible presence of one or more tree nuts and 0/18 samples without cashew declared on the label in any manner. The cashew ELISA can be used to detect undeclared cashew residue in foods and as a potential tool for the food industry to assess the effectiveness of allergen control strategies and to guarantee compliance with food labeling regulatory requirements.

  10. Assessment of natural variability of maize lipid transfer protein using a validated sandwich ELISA

    Science.gov (United States)

    Lipid transfer protein (LTP) is the main causative agent for rare food allergic reactions to maize. This report describes a new, validated ELISA that accurately measures LTP concentrations from 0.2 to 6.4 ng/ml. The levels of LTP ranged from 171 to 865 µg/g grain, a 5.1 fold differences, across a ...

  11. Development and validation of sensitive sandwich ELISAs for two investigational nonapeptide metastin receptor agonists, TAK-448 and TAK-683.

    Science.gov (United States)

    Yoshida, Nobuyo; Nishizawa, Naoki; Matsui, Hisanori; Moriya, Yuu; Kitada, Chieko; Asami, Taiji; Matsumoto, Hirokazu

    2012-11-01

    TAK-448 and TAK-683, investigational agents with potential utility in the treatment of prostate cancer, are potent low molecular weight metastin receptor agonists consisting of nine amino acids. Monoclonal antibodies (mAbs) against these agents were developed to facilitate their evaluation in preclinical studies. Six mAbs were obtained from four immunogens. Three mAbs recognized the C-terminal of TAK-683 and TAK-448, two recognized the N-terminal of TAK-683, and one recognized the N-terminal of TAK-448. Using various combinations of these six mAbs, sandwich ELISAs for TAK-448 and TAK-683 were developed. These assays were highly sensitive, specific, and accurate. The detection limit for TAK-448 and TAK-683 was 3 and 5 pg/mL, respectively, and there was no interference from rat plasma, rat metastin, or analogs of TAK-448/TAK-683. Recovery achieved ≤±10% with intra-/inter-day assay precision coefficient of variation <10%. The assay demonstrated high stability and sample pre-treatment was not required. Each assay detected the dose-dependent concentration of TAK-448 and TAK-683 in blood 24h after a single intravenous administration of 0.1 and 1mg/kg doses. In conclusion, sensitive sandwich ELISAs were developed to detect the small peptides TAK-448 and TAK-683. The novel assays reliably quantified these nonapeptides in rat plasma, and thus will be useful for preclinical studies of these agents. This methodology may be applicable to the development of similar assays for other short peptides.

  12. Development and optimization of a double antibody sandwich ELISA for the detection of goose T cell surface CD8α molecule

    Institute of Scientific and Technical Information of China (English)

    ZHANG Wei; YANG Qiao; WU Ying; CHEN Xiao-yue; CHENG An-chun; CHENG Bei-bei; CHEN Shun; WANG Ming-shu; JIA Ren-yong; ZHU De-kang; LIU Ma-feng; LIU Fei; SUN Kun-feng

    2016-01-01

    CD8, a glycoprotein on the surface of T cels, is involved in the defense against viral infection and plays signiifcant roles in antigen presentation and in the antiviral immune response. CD8 is composed of two chains. Of these, the CD8α chain was chosen for the detection because it involved in both the CD8αα homodimer and the CD8αβ heterodimer. Here, we established a double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) for speciifc detection of goose CD8α (goCD8α). The results showed that the optimal coated antibody and antigen dilutions were 1:50 (the antibody titer was 1:12800) and 1:32 (0.3 ng mL–1), respectively, while the optimal capture antibody and horseradish peroxidase (HRP)-la-beled goat anti-rabbit IgG dilutions were 1:50 (the antibody titer was 1:51200) and 1:4000 (the antibody titer was 1:5000), respectively. The optimal blocking buffer was 5% bovine serum albumin (BSA). The best incubating condition was overnight at 4°C, the best blocking time was 120 min and the best anti-capture antibody working time was 150 min. In addition, the minimum dose detectable by DAS-ELISA was 5×10–3 ng mL–1. Most importantly, goCD8α expression levels in goose spleen mononuclear cels (MNCs) post-Goose parvoviruse (GPV) infection were found to be signiifcantly up-regulated using the DAS-ELISA method, which was consistent with previous results obtained using real-time quantitative PCR. In conclusion, the DAS-ELISA method reported here is a novel, speciifc technique for the clinical detection of goCD8α.

  13. Optimisation of sandwich ELISA based on monoclonal antibodies for the specific measurement of pregnancy-associated plasma protein (PAPP-A) in acute coronary syndrome

    DEFF Research Database (Denmark)

    Rossen, Marie; Iversen, Kasper; Teisner, Ane

    2007-01-01

    OBJECTIVES: PAPP-A has become the principal biochemical serum marker in first trimester screening for Down syndrome, the original data being based on results of a radioimmunoassay (RIA). Recent observations using sandwich ELISA technology have proposed PAPP-A as a potential marker in patients wit...

  14. A comparison of FRP-sandwich penetrating impact test methods

    Energy Technology Data Exchange (ETDEWEB)

    Hildebrand, M. [VTT Manufacturing Technology, Espoo (Finland). Maritime and Mechanical Engineering

    1996-12-31

    The main objective of this project is to identify the test methods which provide useful results for the different types of penetrating impacts occurring in sandwich structures. A series of penetrating impact tests on FRP-sandwich panels is performed using three different test methods and the results of the test methods are compared. The test methods used are the standardised method ISO 6603 and two non-standardised methods. The first non-standardised method uses a pyramid-shaped impactor instead of the cylindrical impactor used in the ISO 6603 method. In the second non-standardised method, the impact test is performed quasistatically using a cylindrical impactor. Possible stages of failure occurring in FRP-sandwich during a penetrating impact are illustrated. A comprehensive test method should be able to provoke various failure modes, as observed in impact failures of actual sandwich structures. The results obtained with the three test methods lead to a different ranking in impact strength of the panels. Hence, impact test results obtained with different test methods are not even qualitatively comparable. The pyramid-shaped impactor is able to generate clearly more failure modes than the cylindrical impactor in the ISO 6603 method. Therefore, it is considered to be of more practical value for determining the impact strength of PRP-sandwich structures. (orig.) (15 refs.)

  15. Production of mono- and polyclonal antibodies to Citrus leprosis virus C2 and their application in triple antibody sandwich ELISA and immunocapture RT-PCR diagnostic assays.

    Science.gov (United States)

    Choudhary, Nandlal; Roy, Avijit; Leon, M G; Wei, G; Nakhla, M K; Levy, L; Brlansky, R H

    2017-05-01

    The newly discovered Citrus leprosis virus cytoplasmic type 2 (CiLV-C2) is one of the causal virus of citrus leprosis disease complex; which leads to substantial loss of citrus production in the states of Meta and Casanare of Colombia. Specific and sensitive detection methods are needed to monitor the dissemination of CiLV-C2 in Colombia, and to prevent introduction of CiLV-C2 to other citrus growing countries. Toward this end, putative coat protein gene (CPG) of CiLV-C2 was amplified from CiLV-C2 infected citrus tissues. The CPG was cloned, expressed and purified a recombinant coat protein of ∼31kDa which used to generate monoclonal antibodies and polyclonal antisera. Four monoclonal antibodies and two polyclonal antisera were selected as being specific following Western blotting. The monoclonal antibody MAb E5 and polyclonal antiserum PAb UF715 were selected testing with an extract of CiLV-C2 infected leaves using triple antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA). In addition, an immunocapture RT-PCR was standardized using MAb E5 for specific and sensitive detection of CiLV-C2. The standardized TAS-ELISA and IC-RT-PCR were able to detect CiLV-C2 in the extracts of symptomatic citrus leprosis tissues up to the dilutions of 1:160 and 1:2580, respectively. Result demonstrated that CiLV-C2 is present in citrus orchards in Meta and Casanare citrus growing areas of Colombia. TAS-ELISA could be used for routine detection of CiLV-C2, epidemiological studies, and for border inspections for quarantine purposes. IC-RT-PCR could be valuable for CiLV-C2 validation and viral genome analysis.

  16. Development and validation of a direct homologous quantitative sandwich ELISA for fathead minnow (Pimephales promelas) vitellogenin.

    Science.gov (United States)

    Eidem, Janne K; Kleivdal, Hans; Kroll, Kevin; Denslow, Nancy; van Aerle, Ronny; Tyler, Charles; Panter, Grace; Hutchinson, Tom; Goksøyr, Anders

    2006-06-15

    . Chem. 75, 2343-2353]. This paper describes the development and validation of a new, homologous enzyme-linked immunosorbent assay (ELISA) for quantification of Vtg in this fish species.

  17. Quantitative Detection of the Foot-And-Mouth Disease Virus Serotype O 146S Antigen for Vaccine Production Using a Double-Antibody Sandwich ELISA and Nonlinear Standard Curves.

    Directory of Open Access Journals (Sweden)

    Xia Feng

    Full Text Available The efficacy of an inactivated foot-and-mouth disease (FMD vaccine is mainly dependent on the integrity of the foot-and-mouth disease virus (FMDV particles. At present, the standard method to quantify the active component, the 146S antigen, of FMD vaccines is sucrose density gradient (SDG analysis. However, this method is highly operator dependent and difficult to automate. In contrast, the enzyme-linked immunosorbent assay (ELISA is a time-saving technique that provides greater simplicity and sensitivity. To establish a valid method to detect and quantify the 146S antigen of a serotype O FMD vaccine, a double-antibody sandwich (DAS ELISA was compared with an SDG analysis. The DAS ELISA was highly correlated with the SDG method (R2 = 0.9215, P<0.01. In contrast to the SDG method, the DAS ELISA was rapid, robust, repeatable and highly sensitive, with a minimum quantification limit of 0.06 μg/mL. This method can be used to determine the effective antigen yields in inactivated vaccines and thus represents an alternative for assessing the potency of FMD vaccines in vitro. But it still needs to be prospectively validated by analyzing a new vaccine preparation and determining the proper protective dose followed by an in vivo vaccination-challenge study to confirm the ELISA findings.

  18. Quantitative Detection of the Foot-And-Mouth Disease Virus Serotype O 146S Antigen for Vaccine Production Using a Double-Antibody Sandwich ELISA and Nonlinear Standard Curves.

    Science.gov (United States)

    Feng, Xia; Ma, Jun-Wu; Sun, Shi-Qi; Guo, Hui-Chen; Yang, Ya-Min; Jin, Ye; Zhou, Guang-Qing; He, Ji-Jun; Guo, Jian-Hong; Qi, Shu-yun; Lin, Mi; Cai, Hu; Liu, Xiang-Tao

    2016-01-01

    The efficacy of an inactivated foot-and-mouth disease (FMD) vaccine is mainly dependent on the integrity of the foot-and-mouth disease virus (FMDV) particles. At present, the standard method to quantify the active component, the 146S antigen, of FMD vaccines is sucrose density gradient (SDG) analysis. However, this method is highly operator dependent and difficult to automate. In contrast, the enzyme-linked immunosorbent assay (ELISA) is a time-saving technique that provides greater simplicity and sensitivity. To establish a valid method to detect and quantify the 146S antigen of a serotype O FMD vaccine, a double-antibody sandwich (DAS) ELISA was compared with an SDG analysis. The DAS ELISA was highly correlated with the SDG method (R2 = 0.9215, P<0.01). In contrast to the SDG method, the DAS ELISA was rapid, robust, repeatable and highly sensitive, with a minimum quantification limit of 0.06 μg/mL. This method can be used to determine the effective antigen yields in inactivated vaccines and thus represents an alternative for assessing the potency of FMD vaccines in vitro. But it still needs to be prospectively validated by analyzing a new vaccine preparation and determining the proper protective dose followed by an in vivo vaccination-challenge study to confirm the ELISA findings.

  19. Protein unfolding allows use of commercial antibodies in an apolipoprotein M sandwich ELISA

    DEFF Research Database (Denmark)

    Bosteen, Markus Høybye; Dahlbäck, Björn; Nielsen, Lars Bo;

    2015-01-01

    diseases such as multiple sclerosis and rheumatoid arthritis. The ability to accurately measure apoM is crucial for investigating its biological functions and possible clinical implications. However, reliable commercial methods have been lacking so far. Therefore, we have developed an assay...

  20. Methods for Using Durable Adhesively Bonded Joints for Sandwich Structures

    Science.gov (United States)

    Smeltzer, Stanley S., III (Inventor); Lundgren, Eric C. (Inventor)

    2016-01-01

    Systems, methods, and apparatus for increasing durability of adhesively bonded joints in a sandwich structure. Such systems, methods, and apparatus includes an first face sheet and an second face sheet as well as an insert structure, the insert structure having a first insert face sheet, a second insert face sheet, and an insert core material. In addition, sandwich core material is arranged between the first face sheet and the second face sheet. A primary bondline may be coupled to the face sheet(s) and the splice. Further, systems, methods, and apparatus of the present disclosure advantageously reduce the load, provide a redundant path, reduce structural fatigue, and/or increase fatigue life.

  1. Methods for Assessing Honeycomb Sandwich Panel Wrinkling Failures

    Science.gov (United States)

    Zalewski, Bart F.; Dial, William B.; Bednarcyk, Brett A.

    2012-01-01

    Efficient closed-form methods for predicting the facesheet wrinkling failure mode in sandwich panels are assessed. Comparisons were made with finite element model predictions for facesheet wrinkling, and a validated closed-form method was implemented in the HyperSizer structure sizing software.

  2. 基于IgY的ELISA用于囊尾蚴循环抗原的检测%Detecting the circulating antigen(CA) of Taenia solium cysticercosis with specific egg yolk antibody (IgY) by sandwich ELISA

    Institute of Scientific and Technical Information of China (English)

    刘玉; 王元伦; 唐雨德

    2013-01-01

    Objective To develop a sensitive and specific double antibody sandwich enzyme-linked immunosorbent assay (ELISA) to detect circulating antigen (CA) of Taeniasoliumcysticercosis with chicken egg yolk immunoglobulin antibodies (IgY).Methods Hens were subcutaneously immunized with CA and the crude IgY was extracted from egg yolk by water dilution method.A sandwich ELISA had been developed by purified IgY antibodies as capture antibody and monoclonal antibodies labeled with peroxidase as detecting antibody.The detection limits of CA were analyzed.The sera and cerebrospinal fluid of patients,the sera of healthy people,sick pigs and healthy pigs were detected in parallel by the established ELISA methods.It's sensitivity and specificity were evaluated by comparison with ELISA based monoclonal antibodies.Results The minimal detectable concentration of CA was 8.3 and 13.9 μg/ml by sandwich ELISA based IgY and monoclonal antibodies,respectively.The positive rates of samples from 139 patients,19 cerebrospinal fluid of patients and 222 sick pigs were 100% (139/139),89.5% (17/19) and 100% (222/222) by sandwich ELISA based IgY respectively.The negative rates of samples from 50 healthy people and 20 healthy pigs were 100%.Conclusion The novel double-antibody sandwich ELISA using anti-CA IgY appears to be sensitive and specific for detection the CA of Taenia solium cysticercosis.It is the promising assay for immunodiagnosis of Taenia solium cysticercosis.%目的 建立基于IgY的双抗体夹心ELISA用于囊尾蚴病的诊断.方法 制备并纯化抗囊尾蚴循环抗原(CA)卵黄抗体(IgY),建立以抗CA的IgY为捕获抗体,酶标记抗CA的单克隆抗体1A5为检测抗体的双抗体夹心ELISA法,共检测样品450份,并与捕获抗体和检测抗体均为单克隆抗体的ELISA法比较,验证方法的敏感性、特异性与实用性.结果 成功制备并鉴定了特异性IgY抗体,建立了基于Igy的双抗体夹心ELISA检测体系.IgY-ELISA和双单

  3. 两种方法检测汉坦病毒滴度的比较研究%A Comparison of Double-antibody Sandwich ELISA and Direct Immunofluoresence Asssy for Detecting and Titering Hantavirus

    Institute of Scientific and Technical Information of China (English)

    白露; 叶伟; 于蒙蒙; 张亮; 于澜; 刘梓谕; 吴兴安; 徐志凯; 张芳琳

    2012-01-01

    比较双抗体夹心ELISA法与直接免疫荧光(DFA)法在检测汉坦病毒灵敏度方面的差别.用汉坦病毒76-118株感染Vero-E6细胞,取不同时间点的病毒感染细胞制备细胞爬片或细胞培养物冻融上清,分别用本室制备的抗汉坦病毒单克隆抗体标记荧光素或辣根过氧化物酶( HRP),建立直接免疫荧光法和双抗体夹心ELISA法检测上述细胞中的病毒抗原,并比较两者的检测灵敏度.用空斑形成实验结果作为金标准,以评价两种方法在检测病毒抗原灵敏度方面的差异.结果:用两种检测方法测定不同时间点的汉坦病毒培养物抗原,双抗体夹心ELISA法不仅检出的时间早,而且其病毒滴度均较IFA法高(P<0.01).说明ELISA法检测汉坦病毒滴度的灵敏度较IFA法更高,并且操作简单、可重复性好、便于大批量规模化检测,因此更适用于汉坦病毒的检测.%To compare the sensitivity between double antibody sandwich ELISA and direct immunofluorescence assay ( DFA) for detection of hantavirus, Vero-E6 cells were infected by Hantaan virus strain 76-118. Cell climb ing slices or freeze-thaw cell culture supernatant was prepared and fluorescein labeled anti-hantavirus monoclonal antibody or horseradish peroxidase ( HRP) labeled monoclonal antibody was used to establish method of double-an tibody sandwich ELISA or direct IFA to detect viral antigen in these cells, and plaque reduction nuetralization test was used as control. And then the optimal detection times were identified and the sensitivity of the two methods was compared. It is resulted that virus titer detected by double-antibody sandwich ELISA was obviously higher than that of DFA (P <0. 01) , and the time point when the virus antigen was detected is more earlier in ELISA than in IFA. It is conclused the sensitivity of double-antibody sandwich ELISA was better than that of direct IFA in detecting hantavirus. And the former method was repeatable and easier to

  4. 双抗体夹心ELISA检测人胎儿血红蛋白体系的建立%Establishment of a sandwich-ELISA system for determination of human fetal hemoglobin

    Institute of Scientific and Technical Information of China (English)

    李雪丽; 文李艳; 冯善伟; 钟梅; 刘艳君; 富宁

    2012-01-01

    目的 建立一种定量检测人胎儿血红蛋白(HbF)的双抗体夹心ELISA方法.方法 用抗人HbF单抗作为捕获抗体,加入待测抗原,检测抗体为兔抗人HbF多克隆抗体,最后加入辣根过氧化物酶标记的羊抗兔IgG.结果 本检测体系灵敏度约为0.039 μg/ml,测量范围0.039-24.66 μg/ml,特异性强,重复性好,100份血标本的检测结果与高效液相色谱法结果符合率高.结论 建立了一种可用于检测人外周血HbF的双抗体夹心ELISA方法,有望为临床上诊断某些与HbF有关的疾病提供帮助.%Objective To establish a sandwich ELISA for the determination of fetal hemoglobin (HbF) in human blood. Methods An anti HbF monoclonal antibody was coated on polystyrene microtiter plates followed by blocking with 5% defat milkithen samples for detection were added to the wells,followed by anti HbF polyclonal antibody pre pared, from rabbit; at last 、addition of HRP goat anti rabbit IgG and. Conventional color development were performed. Re suits This sandwich ELISA system can determine HbF specifically,the sensitivity for HbF is 0. 039 μg/ml,the range of detection is 0. 039 - 24. 66 μg/ml. Conclusion This HbF ELISA is applicable for research and observation in clini cal.

  5. Devices, systems, and methods for conducting sandwich assays using sedimentation

    Energy Technology Data Exchange (ETDEWEB)

    Schaff, Ulrich Y; Sommer, Gregory J; Singh, Anup K; Hatch, Anson V

    2015-02-03

    Embodiments of the present invention are directed toward devices, systems, and method for conducting sandwich assays using sedimentation. In one example, a method includes generating complexes on a plurality of beads in a fluid sample, individual ones of the complexes comprising a capture agent, a target analyte, and a labeling agent. The plurality of beads including the complexes may be transported through a density media, wherein the density media has a density lower than a density of the beads and higher than a density of the fluid sample, and wherein the transporting occurs, at least in part, by sedimentation. Signal may be detected from the labeling agents of the complexes.

  6. Standard Test Method for Dimensional Stability of Sandwich Core Materials

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    2002-01-01

    1.1 This test method covers the determination of the sandwich core dimensional stability in the two plan dimensions. 1.2 The values stated in SI units are to be regarded as the standard. The inch-pound units given may be approximate. This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.

  7. Standard Test Method for Shear Fatigue of Sandwich Core Materials

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    2000-01-01

    1.1 This test method covers determination of the effect of repeated shear loads on sandwich core materials. 1.2 The values stated in SI units are to be regarded as the standard. The inch-pound units given may be approximate. 1.3 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.

  8. Detection and confirmation of PPR virus antigen in sheep and goats by sandwich-ELISA and RT-PCR in Andhra Pradesh, India

    Directory of Open Access Journals (Sweden)

    G. Saritha

    2015-06-01

    Full Text Available Peste des petits ruminants (PPR is a highly contagious disease of domestic and wild small ruminants. Rapid and accurate laboratory assay are essential to enable the implementation of appropriate control strategies to restrict the spread of PPR. The present study was designed to detect the PPR virus (PPRV antigen (N-gene in nasal swabs and tissue samples. A total of 195 samples comprising of 138 nasal swabs from PPR suspected sheep (n=72 and goats (n=66, and 57 tissue samples comprising of lymph nodes from dead sheep (n=39 and goats (n=18 were collected from certain parts of Andhra Pradesh. The samples were subjected to sandwich-ELISA followed by RT-PCR for confirmatory diagnosis. In this study, PPRV could be detected in 27.53% (n=38/138 nasal swabs and 49.12% (n=28/57 tissue samples. Data showed that PPRV infection is widespread in the Andhra Pradesh, India.

  9. The Diagnostic Value of ELISA Method for Pertussis in Children

    Directory of Open Access Journals (Sweden)

    O. P. Popova

    2016-01-01

    Full Text Available Because of low effectiveness of laboratory methods for diagnosing pertussis it is important to look for new ways of verification of this infection. The article presents the analysis of the diagnostic value of ELISA method, which involves the identification of antibodies of different isotypes (IgM, IgG, IgA to pertussis toxoid (PT and filamentous haemagglutinin (FHA. The study included 279 children: 114 were under 1 year of age, 165 — older than 1 year. The pertussis was confirmed in 74.3 ± 2.6% of patients by using ELISA method. A significant proportion of seronegative patients (46.1 ± 6.2 per cent was revealed in the group of patients under 1 year. The pattern of production of antibodies in unvaccinated children was different. It depended on the age of the children and timing of illness. A low proportion of diagnostically significant indicators of IgM-antibodies at 2—3 weeks of illness was typical for patients under 1 year of age (e.g. 6.7 ± 6.5% as compared to 20.0 ± 7.9% and 50.0 ± 15.3 — 1—3 and 4—6 years of age. The diagnosis of pertussis in children under 1 year of age was confirmed mainly by the detection of IgG, starting from the 4th week of the disease. In the examination of vaccinated children diagnostically significant levels of IgA and IgG were identified (even in the late stages of the disease. Thus, the results of the analysis show special significance of using ELISA method for the diagnosis of pertussis in vaccinated children.

  10. A New Sandwich ELISA for Quantification of Thymidine Kinase 1 Protein Levels in Sera from Dogs with Different Malignancies Can Aid in Disease Management.

    Science.gov (United States)

    Jagarlamudi, Kiran Kumar; Moreau, Laura; Westberg, Sara; Rönnberg, Henrik; Eriksson, Staffan

    2015-01-01

    Thymidine kinase 1 (TK1) is a DNA precursor enzyme whose expression is closely correlated with cell proliferation and cell turnover. Sensitive serum TK1 activity assays have been used for monitoring and prognosis of hematological malignancies in both humans and dogs. Here we describe the development of a specific sandwich TK1-ELISA for the quantification of TK1 protein levels in sera from dogs with different malignancies. A combination of rabbit polyclonal anti-dog TK1 antibody and a mouse monoclonal anti-human TK1 antibody was used. Different concentrations of recombinant canine TK1 was used as standard. Clinical evaluation of the ELISA was done by using sera from 42 healthy dogs, 43 dogs with hematological tumors and 55 with solid tumors. An established [3H]-dThd phosphorylation assay was used to determine the TK1 activity levels in the same sera. The mean TK1 activities in dogs with hematological tumors were significantly higher than those found in healthy dogs. In agreement with earlier studies, no significant difference was observed in serum TK1 activities between healthy dogs and dogs with solid tumors. However, the mean TK1 protein levels determined by new TK1-ELISA were significantly higher not only in hematological tumors but also in solid tumors compared to healthy dogs (mean ± SD = 1.30 ± 1.16, 0.67 ± 0.55 and 0.27± 0.10 ng/mL, respectively). Moreover, TK1-ELISA had significantly higher ability to distinguish lymphoma cases from healthy based on receiver operating characteristic analyses (area under the curve, AUC, of 0.96) to that of the activity assay (AUC, 0.84). Furthermore, fluctuations in TK1 protein levels during the course of chemotherapy in dogs with lymphoma closely associated with clinical outcome. Overall, the TK1-ELISA showed significant linear correlation with the TK1 activity assay (rs = 0.6, p<0.0001). Thus, the new TK1-ELISA has sufficient sensitivity and specificity for routine clinical use in veterinary oncology.

  11. Entamoeba histolytica: detection of coproantigens by purified antibody in the capture sandwich ELISA Entamoeba histolytica: detecção de coproantígenos por ELISA de captura utilizando anticorpo purificado

    Directory of Open Access Journals (Sweden)

    Haidee Urdaneta

    1994-12-01

    Full Text Available A sensitive and specific Capture Sandwich ELISA (CSE was developed using polyclonal purified rabbit antibodies against three different axenic strains of Entamoeba histolytica: CSP from Brazil and HM1 - IMSS from Mexico, for the detection of coproantigens in fecal samples. Immunoglobulin G (IgG againstis E. histolytica was isolated from rabbits immunized with throphozoites whole extract in two stages: affinity chromatography in a column containing E. histolytica antigens bound to Sepharose 4B was followed by another chromatography in Sepharose antibodies 4B-Protein A. A Capture Sandwich ELISA using purified antibodies was able to detect 70ng of amebae protein, showing a sensitivity of 93% and specificity of 94%. The combination of microscopic examination and CSE gave a concordance and discordance of 93.25% and 6.75%, respectively. It was concluded that CSE is highly specific for the detection of coproantigens of E. histolytica in feces of infected patients, is quicker to perform, easier and more sensitive than microscopic examination.Foi desenvolvido um teste de ELISA de Captura usando anticorpos policlonais purificados obtidos em coelhos contra três diferentes cepas axênicas de Entamoeba histolytica (ICB-CSP and ICB-462 do Brasil e HM1 do México para detecção de coproantígenos em amostras de fezes de indivíduos: a sintomáticos, b assintomáticos, c com outros parasitos intestinais, e d sadios. Imunoglobulina G (IgG contra E. histolytica foi isolada de imune soro de coelho, em duas etapas: cromatografia de afinidade em uma coluna contendo antígenos de E. histolytica unidos à Sepharose 4B, seguido por outra cromatografía em Sepharose 4B Proteína A. O teste de ELISA usando anticorpos purificados, foi capaz de detectar até um só trofozoíto por lâmina ou 70 ng de proteína de ameba por orifício, apresentando uma sensibilidade de 93% e uma especificidade de 94%. A combinação do exame microscópico com o método de ELISA de Captura

  12. Sandwich Structured Composites for Aeronautics: Methods of Manufacturing Affecting Some Mechanical Properties

    Directory of Open Access Journals (Sweden)

    Aneta Krzyżak

    2016-01-01

    Full Text Available Sandwich panels are composites which consist of two thin laminate outer skins and lightweight (e.g., honeycomb thick core structure. Owing to the core structure, such composites are distinguished by stiffness. Despite the thickness of the core, sandwich composites are light and have a relatively high flexural strength. These composites have a spatial structure, which affects good thermal insulator properties. Sandwich panels are used in aeronautics, road vehicles, ships, and civil engineering. The mechanical properties of these composites are directly dependent on the properties of sandwich components and method of manufacturing. The paper presents some aspects of technology and its influence on mechanical properties of sandwich structure polymer composites. The sandwiches described in the paper were made by three different methods: hand lay-up, press method, and autoclave use. The samples of sandwiches were tested for failure caused by impact load. Sandwiches prepared in the same way were used for structural analysis of adhesive layer between panels and core. The results of research showed that the method of manufacturing, more precisely the pressure while forming sandwich panels, influences some mechanical properties of sandwich structured polymer composites such as flexural strength, impact strength, and compressive strength.

  13. Fundamental study on reactivities of gluten protein types from wheat, rye and barley with five sandwich ELISA test kits.

    Science.gov (United States)

    Lexhaller, Barbara; Tompos, Christine; Scherf, Katharina Anne

    2017-12-15

    Monitoring the compliance of gluten-free foods to the regulatory threshold of 20mg/kg of gluten is essential for celiac disease patients. The different enzyme-linked immunosorbent assays (ELISAs) for gluten detection each have specific characteristics, but there are only a few systematic comparisons. This fundamental study compared the specificities and sensitivities of the R5, G12 and Skerritt monoclonal and two polyclonal antibodies to well-defined gluten protein types (GPT) isolated from wheat, rye and barley flours. Quantitation of protein concentrations by reversed-phase high-performance liquid chromatography provided independent reference values. The ELISA responses showed high variability depending on the type of cereal, the GPT and the antibody used. Overall, ω1,2-gliadins and γ-75k-secalins were most reactive, whereas ω5-gliadins and γ-, B- and D-hordeins were detected with the lowest sensitivities. These results revealed which GPT each antibody is most sensitive to and provided novel insights that will be helpful for appropriate calibration of ELISAs. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Study of Immunoassay Methods for Recombinant Human Erythropoietin(rhEPO)Using Competitive ELISA

    Institute of Scientific and Technical Information of China (English)

    Jin YAN; Jie Bo MI; Wen Bao CHANG

    2004-01-01

    Two different immunoassay methods, competitive indirect enzyme-linked immuno- sorbent assay (CI-ELISA) and amplificative competitive indirect ELISA (ACI-ELISA) using biotin-avidin complex system were studied to detect rhEPO.The linear ranges were 50-20000 ng/mL and 10-50000 ng/mL for CI-ELISA and ACI-ELISA, respectively.The low detection limits of CI-ELISA and ACI-ELISA were 62.8 ng/mL and 8.5 ng/mL, respectively.

  15. Screening methods in alveolar echinococcosis: a follow-up study comparing Emc- and Emf-ELISA with Em2plus-ELISA and ultrasonography.

    Science.gov (United States)

    Hänle, M M; Banzhaf, H-M; Forsbach-Birk, V; Kirch, A; Akinli, A S; Mason, R A; Reuter, S; Kratzer, W

    2009-01-01

    The purpose of the present study was to identify Echinococcus multilocularis infection in follow-up of 95 subjects initially seropostive by Emc-ELISA or Emf-ELISA antibody assays and to compare the utility of these assays with specific Em2plus-ELISA and ultrasound screening for E. multilocularis infection. At follow-up seven subjects were seropositive with both methods, while three were seropositive only with Emc-ELISA and 11 only with Emf-ELISA. All subjects were seronegative with Em2plus-ELISA. There were no manifestations of E. multilocularis infestation by ultrasonographic screening. Seropositivity on Emc-ELISA and Emf-ELISA screening tests does not appear to correlate with manifest alveolar echinococcosis identified by ultrasound. A recommendation for further follow-up of subjects found to be seropositive with Emc-ELISA and Emf-ELISA but with no sonographic evidence of disease is not justified at this time.

  16. [ELISA method for the determination of factor VII antigen].

    Science.gov (United States)

    Jorquera, J I; Aznar, J A; Monteagudo, J; Montoro, J M; Casaña, P; Pascual, I; Bañuls, E; Curats, R; Llopis, F

    1989-12-01

    The low plasma concentration of clotting factor VII makes it difficult to assay its antigenic fraction by the conventional methods of precipitation with specific antigens. Simple and peroxidase-conjugated antisera are currently available from commercial sources, thus allowing one to determine F VII:Ag by enzyme immunoassay. An ELISA method has been developed in this laboratory which provides sensitivity limits about 0.1% of the plasma concentration of F VII and correlates significantly with its functional activity (r = 0.603, n = 44, p less than 0.001). This technique can be highly helpful in characterising molecular variants of F VII, as well as in detecting acquired deficiencies of this factor.

  17. Standard Test Method for Laboratory Aging of Sandwich Constructions

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    1999-01-01

    1.1 This test method covers the determination of the resistance of sandwich panels to severe exposure conditions as measured by the change in selected properties of the material after exposure. The exposure cycle to which the specimen is subjected is an arbitrary test having no correlation with natural weathering conditions. 1.2 The values stated in SI units are to be regarded as the standard. The inch-pound units given may be approximate. 1.3 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.

  18. 测定脑膜炎球菌疫苗C群多糖的双抗体夹心ELISA法的建立%Establishment of a double antibody sandwich ELISA for determination of group C polysaccharide in meningococcal polysaccharide vaccine

    Institute of Scientific and Technical Information of China (English)

    潘殊男; 肖詹蓉; 王宇星; 张霖阳; 史雨舟; 王玲; 张萍

    2012-01-01

    Objective To establish a double antibody sandwich ELISA for determination of group C polysaccharide in groups A,C,Y,W135 meningococcal polysaccharide vaccine (MPV4).Methods The prepared polyclonal antibodies against group C polysaccharide were purified by octanoic acid-ammonium sulfate precipitation method,and then labeled with horseradish peroxidase by sodium periodate method.The polyclonal antibodies against group C polysaccharide were used as coated antibodies and the second enzyme-labled antibodies to establish a double antibody sandwich ELISA.The reaction conditions of the established ELISA were optimized,and specific quantitative determination of group C polysaccharide was carried out by the established ELISA.Results The specificity of the double antibody sandwich ELISA for determination of group C polysaccharide was high,and no cross reactions with groups A,Y and W135 were detected.The best linearity in dose-response curve of group C polysaccharide was found in a range of 2.5-20.0 ng/ml (r > 0.99).The precision and accuracy of the established ELISA were good.Coefficients of variation and recovery rates of intraand inter-assay were 0.6%-9.1% and 87.5%-100.0%,respectively.Quantitation limit was identified as 4.0 ng/ml.Group C polysaccharide contents,molecular dimension KDvalue and recovery rates of three batches of MPV4 detected by the established ELISA were in accordance with those of previous determination and temporary quality control standards.Conclusion The double antibody sandwich ELISA can be applied to detecting the key quality indexes of group C polysaccharide in MPV4.%目的 建立测定A、C、Y、W135群脑膜炎球菌多糖疫苗(groups A,C,Y,W135 meningococcal polysaccharide vaccine,MPV4)C群多糖含量的双抗体夹心ELISA法.方法 制备抗C群多糖多克隆抗体,所得的多克隆抗体经辛酸-硫酸铵沉淀法纯化后,用过碘酸钠法对其标记辣根过氧化物酶.分别将抗C群多糖多克隆抗体作为包被抗

  19. Development and application of a fecal antigen diagnostic sandwich ELISA for estimating prevalence of Fasciola gigantica in cattle in central Java, Indonesia.

    Science.gov (United States)

    Estuningsih, Endah; Spithill, Terry; Raadsma, Herman; Law, Ruby; Adiwinata, G; Meeusen, Els; Piedrafita, David

    2009-04-01

    The purpose of this study was to compare the sensitivity and specificity of an ELISA test to detect Fasciola gigantica antigens (coproantigens) in bovine feces, with fecal egg counting and an ELISA for detecting anti-F. gigantica antibodies in serum. Monoclonal antibodies to cathepsin L were generated and used to capture this antigen in feces of infected cattle. Blood, feces, and livers were collected from 150 cattle at an abattoir in Jakarta, Indonesia, for anti-Fasciola antibodies, coproantigen detection, and F. gigantica egg and worm counts. Fluke recovery varied from 1 to 426 per host, with a mean of 32 flukes. The results showed that the sensitivity and specificity of coproantigen detecting ELISA (95 and 91%, respectively) was better than the anti-F. gigantica antibody ELISA (91 and 88%, respectively) and to fecal egg counting (87 and 100%, respectively). The coproantigen ELISA was able to detect 100% of the cattle with >15 flukes. A survey of 305 cattle in central Java over a 10-mo period validated this test in the field, demonstrating a high prevalence of fascioliasis and establishing the test as a useful diagnostic method to determine patent F. gigantica infections in cattle.

  20. A sandwich ELISA for porcine alpha-1 acid glycoprotein (pAGP, ORM-1) and further demonstration of its use to evaluate growth potential in newborn pigs.

    Science.gov (United States)

    Caperna, T J; Shannon, A E; Stoll, M; Kahl, S; Blomberg, L A; Vallet, J L; Ramsay, T G

    2017-07-01

    A simple, reproducible sandwich, ELISA was developed to measure porcine alpha-1 acid glycoprotein (pAGP, ORM-1) in pig plasma. Porcine AGP isolated from serum was purchased and a polyclonal antisera was prepared in rabbits using the whole pAGP molecule as immunogen. The antiserum was affinity purified, and a portion of the purified antibody fraction was labeled with horseradish peroxidase. Porcine AGP protein was used as a standard, whereas commercially available buffers and reagents were utilized throughout the assay. The assay was specific for pAGP, had a lower limit of detection of 3.2 ng/mL, and could be used to quantify pAGP in plasma or serum. Using this ELISA, we corroborated our previous findings obtained by RID assay, which demonstrated that the AGP concentration in newborn piglets is negatively associated with preweaning growth rate. The current data were obtained using piglets from a different geographical location and genetic background and showed that elevated AGP at birth was associated with reduced preweaning growth rate (P < 0.001, r = 0.433, n = 19 litters). In addition, litters with a greater average AGP at birth were at a growth disadvantage compared with litters with reduced average AGP plasma concentrations (P < 0.001, r = 0.708, n = 19 litters). Litter average plasma AGP was a better predictor of litter preweaning growth rate than average litter birth weight. The data represent further support for using perinatal AGP concentrations as a tool to identify potential slower growing pigs and as a plasma biomarker for predicting litter growth rate. Published by Elsevier Inc.

  1. An Optimum Analysis Method of Sandwich Structures Made from Elastic-viscoelastic Materials

    Institute of Scientific and Technical Information of China (English)

    CHEN Ying-bo; XIA Yu; REN Zhi-gang; LU Zhe-an; WANG Er-lei

    2004-01-01

    Due to a viscoelastic damping middle layer,sandwich structures have the capacity of energy consumption.In this paper,we describe the frequency-dependent property of viscoelastic materials using complex modulus model,and iterative modal strain energy method and iterative complex eigenvalue method are presented to obtain frequency and loss factor of sandwich structures.The two methods are effective and exact for the large-scale complex composite sandwich structures.Then an optimum analysis method is suggested to apply to sandwich structures.Finally,as an example,an optimum analysis of a clamped-clamped sandwich beams is conducted,theoretical closed-form solution and numerical predictions are studied comparatively,and the results agree well.

  2. Antigen sandwiched ELISA for detection of total HIV antibodies%检测抗HIV-1/2型抗体的双抗原夹心方法的建立及其试剂盒的研制

    Institute of Scientific and Technical Information of China (English)

    侯俊; 何红霞; 貌盼勇; 洪世雯; 胡燕; 沈宏辉; 杨松涛

    2001-01-01

    Objective To establish a sensitive antigen sandwiched ELISA(AS) for detection of the total antibodies to HIV-1/2. Methods Based on the gene sequence of HIV-1/2 type and its coded amino acid structure,5 polypeptides were synthesized as coated antigens using solid-phase method. These polypeptides were labelled with horseradish peroxidase. And the total antibodies of HIV-1/2 were detected with the same method. Results These reagents were detected by three batches of HIV panel from The National Institute for the Control of Pharmaceutical and Biological Products (NICPBP). The results indicated that the corresponding rate was 100 %. The variant coefficient rate was less than 10 %. A comparison of antigen sandwiched ELISA with indirect ELISA in detection of a panel(20 positive sera and 20 negative sera) from the NICPBP showed that the general coincident rate of indirect ELISA was 92.5 % and the sandwiched system was 100 %. The HIV-AS diagnostic reagent kits have passed the quality examination of NICPBP. A comparison of antigen sandwiched ELISA with Yapei reagents in detection of 90 normal sera and 88 positive sera for HIV-1/2 showed that the coincident rate was 100%. The reagents were stable at 37℃ for 4 days. This indicated that our reagents were highly specific, sensitive and stable. Conclusion Our antigen sandwiched ELISA reagent for total antibodies of HIV has a merit of sensitivity specificity and stability. It can be clinically used for detection of HIV-1/2 infection and in blood bank for the screening of blood donors.%目的建立更敏感的检测人免疫缺陷病毒(HIV)抗体的方法,并研制检测试剂盒。方法根据HIV-1/2型的基因序列及其所编码氨基酸结构,采用固相法合成了HIV-1型的gp41.1、gp41.2、gp120、p24和HIV-2型的gp36五条多肽,混合包被酶标板作为固相抗原。用辣根过氧化物酶标记以上多肽抗原作为标记物,建立检测血清中抗HIV-1/2抗体的双抗原夹心ELISA法。同时,

  3. A new monoclonal antibody (5D3-F7) which recognizes human monocyte-chemotactic protein-1 but not related chemokines. Development of a sandwich ELISA and in situ detection of producing cells.

    Science.gov (United States)

    Peri, G; Milanese, C; Matteucci, C; Ruco, L; Zhou, D; Sozzani, S; Coletta, I; Mantovani, A

    1994-09-14

    Chemokines are a superfamily of structurally related cytokines involved in leukocyte recruitment in normal and neoplastic tissues. The availability of non-cross-reacting reagents specific for each member of the C-C and C-X-C family is important for careful characterization of their in vitro and in vivo production and relevance. Here we describe a novel, highly specific, mAb against monocyte chemotactic protein-1 (MCP-1). The 5D3-F7 mAb (IgG1,kappa) recognizes human recombinant and natural MCP-1 in ELISA, immunoprecipitation and immunoblot analysis. As a source of natural MCP-1 we used the 8387 human sarcoma line which produces spontaneously MCP-1 and responds to TNF with increased expression and release. The 5D3-F7 mAb inhibited the chemotactic activity of MCP-1 for monocytes. Using the 5D3-F7 mAb and a polyclonal rabbit anti-MCP-1 serum, a sandwich ELISA was developed. In both the direct and the sandwich ELISA, the 5D3-F7 mAb recognized human MCP-1, but not the closely related C-C chemokines MCP-1, MCP-2, MCP-3, MIP-1 alpha, and RANTES and the C-X-C chemokines IL-8, gro alpha and NAP-2. In culture supernatants the sensitivity of the sandwich ELISA was approximately equal to 30 pg/ml. The sandwich ELISA permitted detection of MCP-1 in resting or cytokine-stimulated endothelial, mesothelial and Kaposi's sarcoma cells. Preliminary immunohistochemical analysis revealed production of MCP-1 by macrophage-like cells at sites of inflammation. The 5D3-F7 mAb provides a novel, highly specific reagent with which to investigate the in vitro and in vivo production and role of MCP-1.

  4. PMI Foam Cored Sandwich Components Produced by Means of Different Manufacturing Methods

    Institute of Scientific and Technical Information of China (English)

    Leonhard Maier; HU Pei; Herman Seibert

    2006-01-01

    The paper introduced the structural applications with PMI (Polymethacrylimide) foams in sandwich components for rotor craft, launching vehicle and civil aircraft and discuss some typically used manufacturing methods, such as e. g.in-mould pressing, autoclave curing and resin infusion. The advantages of foam-cored sandwich design versus honeycombcored design will be discussed, focussing on manufacturing costs.

  5. Development of a monoclonal antibody against the left wing of ciguatoxin CTX1B: thiol strategy and detection using a sandwich ELISA.

    Science.gov (United States)

    Tsumuraya, Takeshi; Takeuchi, Katsutoshi; Yamashita, Shuji; Fujii, Ikuo; Hirama, Masahiro

    2012-09-01

    Ciguatera fish poisoning (CFP) is a form of food poisoning caused by the ingestion of a variety of reef fish that have accumulated trace amounts of ciguatoxins produced by dinoflagellates of the genus Gambierdiscus through the food chain. CFP affects more than 50,000 people each year. The extremely low level of the causative neurotoxins, ciguatoxins, in fish has hampered the preparation of antibodies for detecting the toxins. In this paper, we describe a thiol strategy for synthesizing a keyhole limpet hemocyanin (KLH)-conjugate (20) of the ABCDE-ring fragment of the Pacific ciguatoxins, CTX1B (1) and 54-deoxyCTX1B (4). We succeeded in producing a monoclonal antibody (3G8) against the left wings of these ciguatoxins by immunizing mice with the hapten-KLH conjugate (20) as the synthetic antigen. The most promising mAb, 3G8, does not cross-react with other related marine toxins. Sandwich enzyme-linked immunosorbent assay (ELISA) utilizing 3G8 and the previously prepared monoclonal antibody (8H4) enabled us to detect 1 specifically at less than 0.28 ng/mL.

  6. Research overview of design method of super light multi-hole class- honeycomb sandwich structure materials

    Directory of Open Access Journals (Sweden)

    Xiang LI

    Full Text Available With the sandwich structure materials' application and promotion in the field of engineering continuously, existing sandwich structure material gradually cannot meet the design requirements. It is very urgent to develop new sandwich structure materials of high efficiency, energy saving and easy to process. The project puts forward and constructs a new kind of class-honeycomb sandwich structure material combined with important application backgrounds that super light and high strength metal sandwich structure materials are applied into the high weight and high energy consumption equipments of automobile, aerospace and machinery and so on. This research involve: mechanical properties equivalent method for the class-honeycomb sandwich structure and its core; Strength, stiffness and inherent frequency characteristic and failure criterions of the class-honeycomb sandwich structure; based on the failure criterions constructing the multiple-constraint models of the class-honeycomb sandwich structure. The research tries to put forward a new method for innovative design of lightweight material and structure and new ideas of lightweight technology research in theory and practice.

  7. 马流感病毒双抗体夹心ELISA检测方法的建立%Development of Double Antibody Sandwich ELISA for Detection of Equine Influenza Virus

    Institute of Scientific and Technical Information of China (English)

    姬媛媛; 郭巍; 王晓钧; 王征; 卢刚; 赵立平; 李红梅; 相文华

    2011-01-01

    To develop a rapid and effective method for Equine influenza virus (EIV) detection,polyclonal antibodies against EIV A/equine/Xinjiang/07 strain and monoclonal antibody against NP of EIV were generated respectively. Then a double antibody sandwich ELISA (DAS-ELISA)was developed. The specificity of the optimized DAS-ELISA was evaluated using EIV, Equine arteritis virus, Equine herpes virus-1, Equine herpes virus-4 and Japanese encephalitis virus, resulting in only EIV specimens yielding a strong signal. Compared with hemagglutination test, its sensitivity was as two point five to ten times as the later. And it had cross-reactivity with H7N7 subtype. Meanwhile it is suitable for detection of virus from the nasal swabs of experimentally infected equines. The results revealed that the ELISA possessed good specificity and higher sensitivity, indicating a suitable method for rapid detection of EIV.%为建立一种快速、有效的检测马流感病毒(Equine influenza virus,EIV)的方法,以EIV中国分离株A/马/新疆/07(H3N8)制备的多克隆抗体为捕获抗体,原核表达的核蛋白(NP)制备的单克隆抗体为检测抗体,在国内首次建立了检测EIV的双抗体夹心ELISA方法.用该检测方法分别检测EIV、马动脉炎病毒、马疱疹病毒1型、马疱疹病毒4型和马乙型脑炎病毒阳性样品.结果表明,该ELISA方法具有良好的特异性;与常规检测EIV的血凝试验相比,其敏感性是后者的2.5~10倍;同时与H7N7亚型EIV有交叉反应.攻毒试验结果表明该方法可有效检测鼻腔分泌物中的EIV.该方法的建立为EIV的检测及早期防控提供了有效工具.

  8. Immunodetection of Triticum mosaic virus by DAS- and DAC-ELISA using antibodies produced against coat protein expressed in Escherichia coli: potential for high-throughput diagnostic methods.

    Science.gov (United States)

    Tatineni, Satyanarayana; Sarath, Gautam; Seifers, Dallas; French, Roy

    2013-04-01

    Triticum mosaic virus (TriMV), an economically important virus infecting wheat in the Great Plains region of the USA, is the type species of the Poacevirus genus in the family Potyviridae. Sensitive and high-throughput serology-based detection methods are crucial for the management of TriMV and germplasm screening in wheat breeding programs. In this study, TriMV coat protein (CP) was expressed in Escherichia coli, and polyclonal antibodies were generated against purified soluble native form recombinant CP (rCP) in rabbits. Specificity and sensitivity of resulting antibodies were tested in Western immuno-blot and enzyme-linked immunosorbent assays (ELISA). In direct antigen coating (DAC)-ELISA, antibodies reacted specifically, beyond 1:20,000 dilution with TriMV in crude sap, but not with healthy extracts, and antiserum at a 1:10,000 dilution detected TriMV in crude sap up to 1:4860 dilution. Notably, rabbit anti-TriMV IgG and anti-TriMV IgG-alkaline phosphatase conjugate reacted positively with native virions in crude sap in a double antibody sandwich-ELISA, suggesting that these antibodies can be used as coating antibodies which is crucial for any 'sandwich' type of assays. Finally, the recombinant antibodies reacted positively in ELISA with representative TriMV isolates collected from fields, suggesting that antibodies generated against rCP can be used for sensitive, large-scale, and broad-spectrum detection of TriMV.

  9. Development of Double Antibody Sandwich ELISA for Vibrio TDH-VVC-VMHA Fusion Protein%弧菌融合蛋白TDH-VVC-VMHA双抗体夹心ELISA检测方法的建立

    Institute of Scientific and Technical Information of China (English)

    刘秀萍; 王铁良; 唐峰

    2011-01-01

    In order to improve the detection speed and efficiency, a double antibody sandwich ELISA was developed to detect Vibrio parahaemolyticus, Vibrio vulnificus or Vibrio minicus in food simultaneously. The cytorrhyctes of the fusion protein were purified by the electroeluting to immunize cavia cobaya, anti-serum were uesed as capture antibody,goat anti-guinea pig IgG/HRP were uesed as labelled-antibody, optimizing condition was determined by use of the chessboard titration method. The results showed that, the double antibody sandwich ELISA having obvious positive reaction with the culture supernatant of Vibrio parahaemolyticus, Vibrio vulnificus, Vibrio mimicus, but no positive reaction with 8 other culture supernatant of contrast strains including Pseudomonas aeruginosa, Staphylococcus aureus, Listeria, etc. It indicated that this method could be used to detect Vibrio parahaemolyticus, Vibrio vulnificus and V. Mimicus toxin in food simultaneously.%为提高食品中副溶血孤菌、创伤弧菌和拟态弧菌毒素初筛的速度与效率,建立了双抗体夹心ELISA检测方法.以融合蛋白TDH-VVC-VMHA包涵体为抗原免疫豚鼠,采用豚鼠抗融合蛋白免疫血清为包被抗体,辣根过氧化物酶(HRP)标记羊抗豚鼠IgG为标记抗体,应用棋盘滴定法确定ELISA的最适条件.结果表明,所建立的方法与副溶血弧菌、创伤弧菌、拟态弧菌的培养物上清有较明显的阳性反应,与绿脓杆菌、金黄色葡菌球菌、李氏杆菌、甲型副伤寒沙门氏菌、猪霍乱沙门氏菌等5种非弧菌属食物中毒菌培养物上清未见交叉反应,与溶藻弧菌、河流弧菌和麦氏弧菌等3种弧菌培养物上清也未见交叉反应.这说明,该方法可以用于食品中副溶血弧菌、创伤弧菌和拟态弧菌毒素的快速同步免疫学检测.

  10. Multiscale Modeling Methods for Analysis of Failure Modes in Foldcore Sandwich Panels

    Science.gov (United States)

    Sturm, R.; Schatrow, P.; Klett, Y.

    2015-12-01

    The paper presents an homogenised core model suitable for use in the analysis of fuselage sandwich panels with folded composite cores under combined loading conditions. Within a multiscale numerical design process a failure criterion was derived for describing the macroscopic behaviour of folded cores under combined loading using a detailed foldcore micromodel. The multiscale modelling method was validated by simulation of combined compression/bending failure of foldcore sandwich panels.

  11. 牛初乳中sIgA双抗夹心ELISA检测方法的建立%Establishment of a sandwich ELISA for quantitive measurement of sIgA

    Institute of Scientific and Technical Information of China (English)

    朱洪艳; 赵红宇; 周盛华; 高学军

    2011-01-01

    为定量检测牛初乳中sIgA,建立双抗夹ELISA方法.以牛初乳中sIgA为抗原,免疫新西兰白兔,制得抗血清,经纯化后作为包被抗体;利用简易过碘酸钠法对兔抗牛sIgA标记辣根过氧化物酶(HTP),制得酶标抗体;通过方阵滴定法确定ELISA最佳工作条件;进行特异性、重复性、稳定性实验并且检测牛初乳粉样本中的sIgA含量.结果:成功建立了一种定量检测牛初乳中sIgA的双抗夹心ELISA方法.该方法线性范围:15.625~1 000μg/L;最低检测限(LOD)为15.625μg/L;精密度为:批内CV=4.8%;批间CV=6.5%.%To establish an effective ELISA mathod for the determination of bovine colostrum sIgA.Rabbit was immunized by bovine colostrum sIgA to obtain the antiserum.Useing the purified antiserum as coating antibody.All the anti- sIgA were labeled with horseradish peroxidase by sodium oxidation method, and take the HRP labeled anti- sIgA for labeled antibody.The optimal concentrations of ELISA were defined by titration.For speciticity reproducibility and sensitivity experiment and measured the sIgA levels of bovine colostrum with this assay.Results a sandwich ELISA assay for detecting sIgA in bovine colostrum was established successfully.The assay result was linear over a concentration range of 15.625~1 000 μg/L with a limit of detection(LOD) of 15.625 μg/L.The coefficients of variation was 4.8% within assay and 6.5% between assay.

  12. Systems, Apparatuses, and Methods for Using Durable Adhesively Bonded Joints for Sandwich Structures

    Science.gov (United States)

    Smeltzer, III, Stanley S. (Inventor); Lundgren, Eric C. (Inventor)

    2014-01-01

    Systems, methods, and apparatus for increasing durability of adhesively bonded joints in a sandwich structure. Such systems, methods, and apparatus includes an first face sheet and an second face sheet as well as an insert structure, the insert structure having a first insert face sheet, a second insert face sheet, and an insert core material. In addition, sandwich core material is arranged between the first face sheet and the second face sheet. A primary bondline may be coupled to the face sheet(s) and the splice. Further, systems, methods, and apparatus of the present disclosure advantageously reduce the load, provide a redundant path, reduce structural fatigue, and/or increase fatigue life.

  13. Comparative detection of bacterial adhesion to Caco-2 cells with ELISA, radioactivity and plate count methods.

    Science.gov (United States)

    Le Blay, Gwenaëlle; Fliss, Ismaïl; Lacroix, Christophe

    2004-11-01

    Different methods are used to study bacterial adhesion to intestinal epithelial cells, which is an important step in pathogenic infection as well as in probiotic colonization of the intestinal tract. The aim of this study was to compare the ELISA-based method with more conventional plate count and radiolabeling methods for bacterial adhesion detection. An ELISA-based assay was optimized for the detection of Bifidobacterium longum and Escherichia coli O157:H7, which are low and highly adherent bacteria, respectively. In agreement with previous investigations, a percentage of adhesion below 1% was obtained for B. longum with ELISA. However, high nonspecific background and low positive signals were measured due to the use of polyclonal antibodies and the low adhesion capacity with this strain. In contrast, the ELISA-based method developed for E. coli adhesion detected a high adhesion percentage (15%). For this bacterium the three methods tested gave similar results for the highest bacterial concentrations (6.8 Log CFU added bacteria/well). However, differences among methods increased with the addition of decreased bacterial concentration due to different detection thresholds (5.9, 5.6 and 2.9 Log CFU adherent bacteria/well for radioactivity, ELISA and plate count methods, respectively). The ELISA-based method was shown to be a good predictor for bacterial adhesion compared to the radiolabeling method when good quality specific antibodies were used. This technique is convenient and allows handling of numerous samples.

  14. Bepaling van het gehalte aan soja in vleesprodukten m.b.v. ELISA-methode

    NARCIS (Netherlands)

    Visser-Meijer, M.A.; Buizer, F.G.

    1983-01-01

    Het gehalte soja-eiwit is bepaald in 3 monsters vleesprodukt met bekende hoeveelheid soja en vervolgens in 7 monsters vleesprodukt met onbekende hoeveelheid soja. De bepaling geschiedde met behulp van een ELISA methode.

  15. Bepaling van het gehalte aan soja in vleesprodukten m.b.v. ELISA-methode

    OpenAIRE

    Visser-Meijer, M.A.; Buizer, F.G.

    1983-01-01

    Het gehalte soja-eiwit is bepaald in 3 monsters vleesprodukt met bekende hoeveelheid soja en vervolgens in 7 monsters vleesprodukt met onbekende hoeveelheid soja. De bepaling geschiedde met behulp van een ELISA methode.

  16. Finite Element Analysis of Bend Test of Sandwich Structures Using Strain Energy Based Homogenization Method

    Directory of Open Access Journals (Sweden)

    Hassan Ijaz

    2017-01-01

    Full Text Available The purpose of this article is to present a simplified methodology for analysis of sandwich structures using the homogenization method. This methodology is based upon the strain energy criterion. Normally, sandwich structures are composed of hexagonal core and face sheets and a complete and complex hexagonal core is modeled for finite element (FE structural analysis. In the present work, the hexagonal core is replaced by a simple equivalent volume for FE analysis. The properties of an equivalent volume were calculated by taking a single representative cell for the entire core structure and the analysis was performed to determine the effective elastic orthotropic modulus of the equivalent volume. Since each elemental cell of the hexagonal core repeats itself within the in-plane direction, periodic boundary conditions were applied to the single cell to obtain the more realistic values of effective modulus. A sandwich beam was then modeled using determined effective properties. 3D FE analysis of Three- and Four-Point Bend Tests (3PBT and 4PBT for sandwich structures having an equivalent polypropylene honeycomb core and Glass Fiber Reinforced Plastic (GFRP composite face sheets are performed in the present study. The authenticity of the proposed methodology has been verified by comparing the simulation results with the experimental bend test results on hexagonal core sandwich beams.

  17. The Conformal Method and the Conformal Thin-Sandwich Method Are the Same

    CERN Document Server

    Maxwell, David

    2014-01-01

    The conformal method developed in the 1970s and the more recent Lagrangian and Hamiltonian conformal thin-sandwich methods are techniques for finding solutions of the Einstein constraint equations. We show that they are manifestations of a single conformal method: there is a straightforward way to convert back and forth between the parameters for these methods so that the corresponding solutions of the Einstein constraint equations agree. The unifying idea is the need to clearly distinguish tangent and cotangent vectors to the space of conformal classes on a manifold, and we introduce a vocabulary for working with these objects without reference to a particular representative background metric. As a consequence of these conceptual advantages, we demonstrate how to strengthen previous near-CMC existence and non-existence theorems for the original conformal method to include metrics with scalar curvatures that change sign.

  18. Development of an enzyme-linked immuno-sorbent assay (ELISA) method for carbofuran residues.

    Science.gov (United States)

    Yang, Jinyi; Wang, Hong; Jiang, Yueming; Sun, Yuanming; Pan, Ke; Lei, Hongtao; Wu, Qing; Shen, Yudong; Xiao, Zhili; Xu, Zhenlin

    2008-04-17

    The haptens 4-[[(2,3-dihydro-2,2-dimethyl-7-benzofuranyloxy)carbonyl]-amino]butanoic acid (BFNB) and 6-[((2,3-dihydro-2,2-dimethyl-7-benzofuranyloxy)-carbonylamino]hexanoic acid (BFNH) were synthesized and then used to develop a rapid,specific and sensitive ELISA method to determine residues of the pesticide carbofuran in a variety of matrices. A hybridoma cell line (5D3) producing anti-carbofuran monoclonal antibodies (MAbs) was also established. Based on the MAbs in combination with the heterologous hapten BFNH coupled to either horseradish peroxidase (HRP) or ovalbumin(OVA), four ELISAs (formats I-IV) for the quantification of carbofuran were developed and compared. Among them, the optimized format II (the conjugate-coated direct competitive ELISA) showed the best characteristics, with an IC50 value of 18.49 ng/mL, a limit of detection of 0.11 ng/mL and the shortest assay time (1 h). This ELISA method was then applied to the determinations of carbofuran in environmental water, soil and food samples. The relative standard deviations (R.S.D.s) ranged from 1.8% to 21.3% and the mean recoveries were 104.6%, 108.3%, 106.3% and 100.1% for water, soil, lettuce and cabbage, respectively. Thus, the ELISA method of format II exhibited the potential to develop commercial ELISA kits for a rapid detection of carbofuran for human health and environmental safety.

  19. Development of an Enzyme-linked Immuno-Sorbent Assay (ELISA Method for Carbofuran Residues

    Directory of Open Access Journals (Sweden)

    Zhenlin Xu

    2008-04-01

    Full Text Available The haptens 4-[[(2,3-dihydro-2,2-dimethyl-7-benzofuranyloxycarbonyl]-amino]butanoic acid (BFNB and 6-[((2,3-dihydro-2,2-dimethyl-7-benzofuranyloxy-carbonylamino]hexanoic acid (BFNH were synthesized and then used to develop a rapid,specific and sensitive ELISA method to determine residues of the pesticide carbofuran in avariety of matrices. A hybridoma cell line (5D3 producing anti-carbofuran monoclonalantibodies (MAbs was also established. Based on the MAbs in combination with theheterologous hapten BFNH coupled to either horseradish peroxidase (HRP or ovalbumin(OVA, four ELISAs (formats I-IV for the quantification of carbofuran were developedand compared. Among them, the optimized format II (the conjugate-coated directcompetitive ELISA showed the best characteristics, with an IC50 value of 18.49 ng/mL, alimit of detection of 0.11 ng/mL and the shortest assay time (1 h. This ELISA method wasthen applied to the determinations of carbofuran in environmental water, soil and foodsamples. The relative standard deviations (R.S.D.s ranged from 1.8% to 21.3% and themean recoveries were 104.6%, 108.3%, 106.3% and 100.1% for water, soil, lettuce andcabbage, respectively. Thus, the ELISA method of format II exhibited the potential todevelop commercial ELISA kits for a rapid detection of carbofuran for human health andenvironmental safety.

  20. Novel strengthening methods for ultralightweight sandwich structures with periodic lattice cores

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    Recent development of ultralightweight lattice-cored sandwiches is reviewed,with focus placed on various novel fabrication methods introduced to strengthen these structures,covering not only research results published in the Science China Series E-Tech Sci,but also those in other domestic and overseas scientific journals.

  1. 水貂肠炎病毒双抗体夹心ELISA检测方法的建立%Establishment of double antibody sandwich ELISA for detection of mink enteritis virus

    Institute of Scientific and Technical Information of China (English)

    王建科; 程世鹏; 易立; 杨莘; 罗彬; 许红丽; 闫喜军; 武华

    2011-01-01

    In order to establish double antibody sandwich enzyme-linked immunosorbent assay(DAS-ELISA) for detection of mink enteritis virus(MEV),anti-MEV monoclonal antibody and rabbit anti-MEV polyclonal antibody were used as the capture antibody and detecting antibody,respectively. The optimal dilution of the capture antibody and the detecting antibody capable of detecting MEV antigens were 1: 20 and 1: 3 200 in the check-board titration respectively. Positive samples with MEV,Aleutian mink disease virus and canine distemper virus were examined by the established ELISA respectively. In result,the developed DAS-ELISA had good specificity. A total of 158 samples were tested both by the developed DAS-ELISA and by polymerase chain reaction(PCR). 40 of the tested samples were positive by the developed ADS-ELISA and 44 by PCR. The specificity and sensitivity of the developed DAS-ELISA were 95.6 %and 79.5 %, respectively. The coincidental rate between the developed DAS-ELISA and PCR was 91.1%. These results indicated that the developed DAS-ELISA was suitable for rapid detection and epidemiological investigation of MEV infection in mink.%以抗水貂肠炎病毒(mink enteritis virus,MEV)单克隆抗体为捕获抗体、兔抗MEV多克隆抗体为检测抗体,建立了MEV双抗体夹心ELISA检测方法.该方法用于MEV抗原的检测.经过试验,单克隆抗体的最适稀释度为1:20,兔抗MEV多克隆抗体的最适稀释度为1:3200.用该ELISA方法分别检测MEV、水貂阿留中病痛毒、犬瘟热病毒样品.结果表明,ELISA方法具有良好的特异性.用该ELISA和PCR同时检测158份临床样品,其中ELISA检测40份为阳性,PCR检测44份为阳性,该ELISA的特异性和敏感性分别为95.6%和79.5%,这2种方法的符合率为91.1%.该方法的建立为MEV的检测及水貂病毒性肠炎的流行病学调查提供了工具.

  2. Evaluation of a Commercial Sandwich Enzyme-Linked Immunosorbent Assay for the Quantification of Beta-Casomorphin 7 in Yogurt Using Solid-Phase Extraction Coupled to Liquid Chromatography-Tandem Mass Spectrometry as the "Gold Standard" Method.

    Science.gov (United States)

    Nguyen, Duc Doan; Busetti, Francesco; Johnson, Stuart Keith; Solah, Vicky Ann

    2017-08-01

    This study investigated beta-casomorphin 7 (BCM7) in yogurt by means of LC-tandem MS (MS/MS) and enzyme-linkedimmunosorbent assay (ELISA) and use LC-MS/MS as the "gold standard" method to evaluate the applicability of a commercial ELISA. The level of BCM7 in milk obtained from ELISA analysis was much lower than that obtained by LC-MS/MS analysis and trended to increase during fermentation and storage of yogurt. Meanwhile, the results obtained from LC-MS/MS showed that BCM7 degraded during stages of yogurt processing, and its degradation may have been caused by X-prolyl dipeptidyl aminopeptidase activity. As a result, the commercial sandwich ELISA kit was not suitable for the quantification of BCM7 in fermented dairy milk.

  3. Sound transmission analysis of partially treated MR fluid-based sandwich panels using finite element method

    Science.gov (United States)

    Hemmatian, M.; Sedaghati, R.

    2017-04-01

    This study aims at developing a finite element model to predict the sound transmission loss (STL) of a multilayer panel partially treated with a Magnetorheological (MR) fluid core layer. MR fluids are smart materials with promising controllable rheological characteristics in which the application of an external magnetic field instantly changes their rheological properties. Partial treatment of sandwich panels with MR fluid core layer provides an opportunity to change stiffness and damping of the structure without significantly increasing the mass. The STL of a finite sandwich panel partially treated with MR fluid is modeled using the finite element (FE) method. Circular sandwich panels with clamped boundary condition and elastic face sheets in which the core layer is segmented circumferentially is considered. The MR fluid core layer is considered as a viscoelastic material with complex shear modulus with the magnetic field and frequency dependent storage and loss moduli. Neglecting the effect of the panel's vibration on the pressure forcing function, the work done by the acoustic pressure is expressed as a function of the blocked pressure in order to calculate the force vector in the equation of the motion of the panel. The governing finite element equation of motion of the MR sandwich panel is then developed to predict the transverse vibration of the panel which can then be utilized to obtain the radiated sound using Green's function. The developed model is used to conduct a systematic parametric study on the effect of different locations of MR fluid treatment on the natural frequencies and the STL.

  4. Development and Preliminary Application of ELISA Method for Diphtheria Toxoid%白喉类毒素ELISA检测方法的建立及初步应用

    Institute of Scientific and Technical Information of China (English)

    刘朝阳; 景辉; 陈艳红; 张斌; 马相虎; 朱金华

    2011-01-01

    目的 建立白喉类毒素双抗体夹心ELISA检测方法,并进行验证及初步应用.方法 以马抗白喉类毒素血清作为包被抗体,HRP标记的白喉絮状反应抗毒素作为酶标抗体,建立白喉类毒素双抗体夹心ELISA检测方法,并对该方法进行重复性、特异性验证、最佳线性范围和检测限确定及初步应用.结果 经验证,该方法重复性好,特异性强,白喉类毒素含量在0.9765~125.0000ng/ml之间时线性关系良好,检测限为7.812 ng/ml,该方法检测了3批白喉类毒素原液,与动物实验结果基本相符.结论 已建立了白喉类毒素双抗体夹心ELISA检测方法,可用于检测白喉类毒素的含量和抗原性.%Objective To develop, verify and preliminarily apply a double antibody sandwich ELISA method for diphtheria toxoid. Methods A double antibody sandwich ELISA method was developed using horse antiserum against diphtheria toxoid as coating antibody and HRP-labeled diphtheria flocculation antitoxin as enzyme labeled antibody, then verified for reproducibility and specificity, determined for optimal linear range and detection limit, and applied preliminarily. Results The developed method showed high reproducibility and specificity, of which the optimal linear range and detection limit were 0. 976 5 ~ 125. 000 0 ng/ml and 7. 812 ng/ml respectively. Three batches of bulk diphtheria toxoid were detected by the developed method, and the results were basically consistent with thoes by animal test. Conclusion A double antibody sandwich ELISA method was developed, which might be used for determination of content and antigenicity of diphtheria toxoid.

  5. 单克隆抗体与多克隆抗体配对ELISA方法比较%Study of the Optimum Pairing of Polyclonal Antibodies and Monoclonal Antibodies with Sandwich ELISA

    Institute of Scientific and Technical Information of China (English)

    张小兵; 邸禄芹; 吴萌; 闫静辉

    2009-01-01

    Using human chorionic gonadotrophin( HCG) as the antigen,polyclonal antibodies and monoclonal antibodies(McAbs) specific for HCG were generated, characterized and highly purified. Then these antibodies were labeled with horseradish peroxidase (HRP). By a double antibody sandwich enzyme-linked immunosorhent assay, several problems of the optimum pairing of polyclonal antibodies and monoclonal antibodies were discussed. The results showed that with monoclonal antibody as trapping antibody and polyclonal antibody labeling with enzyme as testing antibody which should be diluted with animal sera as diluent,the sandwich ELISA had a highly specificity and sensitivity to detect antigen.%以人绒毛膜促性腺激素(HCG)为抗原,制备出对HCG的多克隆抗体和特异性单克隆抗体,并进行抗体纯化和特性分析,利用辣根过氧化物酶(HRP)分别对其进行了标记.采用双抗夹心ELISA试验,探讨了多克隆抗体与单克隆抗体配对的若干事项.结果表明,利用单克隆抗体和酶标多克隆抗体配对,并用含动物血清的稀释液稀释酶标抗体,可实现对检测原的高特异性和高灵敏度检测.

  6. Usefulness of ELISA Methods for Assessing LPS Interactions with Proteins and Peptides

    Science.gov (United States)

    Martínez-Sernández, Victoria; Orbegozo-Medina, Ricardo A.; Romarís, Fernanda; Paniagua, Esperanza; Ubeira, Florencio M.

    2016-01-01

    Lipopolysaccharide (LPS), the major constituent of the outer membrane of Gram-negative bacteria, can trigger severe inflammatory responses during bacterial infections, possibly leading to septic shock. One approach to combatting endotoxic shock is to neutralize the most conserved part and major mediator of LPS activity (lipid A) with LPS-binding proteins or peptides. Although several available assays evaluate the biological activity of these molecules on LPS (e.g. inhibition of LPS-induced TNF-α production in macrophages), the development of simple and cost-effective methods that would enable preliminary screening of large numbers of potential candidate molecules is of great interest. Moreover, it would be also desirable that such methods could provide information about the possible biological relevance of the interactions between proteins and LPS, which may enhance or neutralize LPS-induced inflammatory responses. In this study, we designed and evaluated different types of ELISA that could be used to study possible interactions between LPS and any protein or peptide. We also analysed the usefulness and limitations of the different ELISAs. Specifically, we tested the capacity of several proteins and peptides to bind FITC-labeled LPSs from Escherichia coli serotypes O111:B4 and O55:B5 in an indirect ELISA and in two competitive ELISAs including casein hydrolysate (hCAS) and biotinylated polymyxin B (captured by deglycosylated avidin; PMX) as LPS-binding agents in the solid phase. We also examined the influence of pH, detergents and different blocking agents on LPS binding. Our results showed that the competitive hCAS-ELISA performed under mildly acidic conditions can be used as a general method for studying LPS interactions, while the more restrictive PMX-ELISA may help to identify proteins/peptides that are likely to have neutralizing properties in vitro or in vivo. PMID:27249227

  7. Sandwich Panels

    Directory of Open Access Journals (Sweden)

    N. Ramachandran

    1963-05-01

    Full Text Available This introductory article give an insight into the different methods employed in the construction of Sandwich panels, their limitations and future design application for defence use as a structural element with one of the highest strength-weight ratios yet devised.

  8. Preparation of monoclonal antibodies against bovine viral diarrhea virus and the double antibody sandwich ELISA develpoment%抗牛病毒性腹泻病毒单克隆抗体的制备及双抗夹心ELISA检测方法的建立

    Institute of Scientific and Technical Information of China (English)

    蒋颖; 朱国强; 林燕清; 陶洁; 孟祥升; 段小丽; 王娟; 王银; 张信军; 王建业

    2012-01-01

    为建立牛病毒性腹泻病毒(BVDV)双抗夹心ELISA检测方法,本研究将BVDV 890病毒株(BVDV-2)浓缩并纯化后免疫BALB/c小鼠,经常规技术进行细胞融合并筛选得到一株稳定分泌IgG的杂交瘤细胞株,命名为3F9.以BVDV单克隆抗体(MAb) IgM为捕获抗体,生物素标记的3F9为检测抗体,初步建立BVDV特异的双抗体夹心ELISA检测方法(BAS-ELISA).采用建立的BAS-ELISA方法检测35份临床病牛血清样品,检出阳性样本13份;与RT-PCR的符合率达到94.29%;表明本研究所建立的BAS-ELISA方法可用于BVDV感染的临床诊断,为BVDV的免疫学研究奠定了基础.%To establish a double monoclonal antibody (MAb)-mediated sandwich ELISA (DAS-ELISA) for detection of bovine viral diarrhea virus (BVDV), the MAb 3D8 (IgM) and biotin-labelled 3F9 (IgG) were used as capture antibody and detection antibody respectively after obtaining a hybridoma cell line stably secreting antibody (3F9). Thirty-five clinically suspected sera samples were detected by this method and the results of 13 samples were positive. Compared with RT-PCR in the paralled experiment with 11 samples positive, the coincidence rate was 94.25%. It is suggested that the DAS-ELISA was a potentially valuable method with high sensitivity and specificity against BVDV.

  9. Sequential chemotherapy and radiotherapy in the sandwich method for advanced endometrial cancer: a meta-analysis.

    Science.gov (United States)

    Gao, Huiqiao; Zhang, Zhenyu

    2015-04-01

    Endometrial cancer is one of the most common gynecological malignancies and the standard treatment modality has not been established.To assess the efficacy and tolerability of a sandwich method consisted of chemotherapy followed by involved field irradiation and additional chemotherapy for the treatment of advanced endometrial cancer.The Medline, Embase, Cochrane, and China National Knowledge Infrastructure (CNKI) Library were searched to identify the relevant literature published between 1970 and September 2014. A meta-analysis was performed to evaluate progression-free survival (PFS), overall survival (OS), and toxicity.A total of 5 articles were subjected to this meta-analysis. The pooled 3-year PFS and OS of patients with advanced endometrial cancer treated with the "sandwich" method was 68% (95% CI: 0.60-0.77) with no heterogeneity (I = 0.00%, P = 0.77) among the studies and 75% (95% CI: 0.61-0.89) with significant heterogeneity (I = 71.8%, P = 0.01), respectively. Pooled analysis of toxicity was not performed because of the substantial heterogeneity.Sequential chemotherapy and radiotherapy in the sandwich method is both efficacious and well tolerated. Large-scale randomized controlled trials (RCTs) are necessary in the future.

  10. Sensitivity improvement of a sandwich-type ELISA immunosensor for the detection of different prostate-specific antigen isoforms in human serum using electrochemical impedance spectroscopy and an ordered and hierarchically organized interfacial supramolecular architecture.

    Science.gov (United States)

    Gutiérrez-Zúñiga, Gabriela Guadalupe; Hernández-López, José Luis

    2016-01-01

    A gold millielectrode (GME) functionalized with a mixed (16-MHA + EG3SH) self-assembled monolayer (SAM) was used to fabricate an indirect enzyme-linked immunosorbent assay (ELISA) immunosensor for the sensitive detection of prostate-specific antigen (PSA), a prostate cancer (PCa) biomarker, in human serum samples. To address and minimize the issue of non-specific protein adsorption, an organic matrix (amine-PEG3-biotin/avidin) was assembled on the previously functionalized electrode surface to build up an ordered and hierarchically organized interfacial supramolecular architecture: Au/16-MHA/EG3SH/amine-PEG3-biotin/avidin. The electrode was then exposed to serum samples at different concentrations of a sandwich-type immunocomplex molecule ((Btn)Ab-AgPSA-(HRP)Ab), and its interfacial properties were characterized using electrochemical impedance spectroscopy (EIS). Calibration curves for polarization resistance (RP) and capacitance (1/C) vs. total and free PSA concentrations were obtained and their analytical quality parameters were determined. This approach was compared with results obtained from a commercially available ELISA immunosensor. The results obtained in this work showed that the proposed immunosensor can be successfully applied to analyze serum samples of patients representative of the Mexican population.

  11. Virtual Design Method for Controlled Failure in Foldcore Sandwich Panels

    Science.gov (United States)

    Sturm, Ralf; Fischer, S.

    2015-12-01

    For certification, novel fuselage concepts have to prove equivalent crashworthiness standards compared to the existing metal reference design. Due to the brittle failure behaviour of CFRP this requirement can only be fulfilled by a controlled progressive crash kinematics. Experiments showed that the failure of a twin-walled fuselage panel can be controlled by a local modification of the core through-thickness compression strength. For folded cores the required change in core properties can be integrated by a modification of the fold pattern. However, the complexity of folded cores requires a virtual design methodology for tailoring the fold pattern according to all static and crash relevant requirements. In this context a foldcore micromodel simulation method is presented to identify the structural response of a twin-walled fuselage panels with folded core under crash relevant loading condition. The simulations showed that a high degree of correlation is required before simulation can replace expensive testing. In the presented studies, the necessary correlation quality could only be obtained by including imperfections of the core material in the micromodel simulation approach.

  12. Validation procedures for quantitative gluten ELISA methods: AOAC allergen community guidance and best practices.

    Science.gov (United States)

    Koerner, Terry B; Abbott, Michael; Godefroy, Samuel Benrejeb; Popping, Bert; Yeung, Jupiter M; Diaz-Amigo, Carmen; Roberts, James; Taylor, Steve L; Baumert, Joseph L; Ulberth, Franz; Wehling, Paul; Koehler, Peter

    2013-01-01

    The food allergen analytical community is endeavoring to create harmonized guidelines for the validation of food allergen ELISA methodologies to help protect food-sensitive individuals and promote consumer confidence. This document provides additional guidance to existing method validation publications for quantitative food allergen ELISA methods. The gluten-specific criterion provided in this document is divided into sections for information required by the method developer about the assay and information for the implementation of the multilaboratory validation study. Many of these recommendations and guidance are built upon the widely accepted Codex Alimentarius definitions and recommendations for gluten-free foods. The information in this document can be used as the basis of a harmonized validation protocol for any ELISA method for gluten, whether proprietary or nonproprietary, that will be submitted to AOAC andlor regulatory authorities or other bodies for status recognition. Future work is planned for the implementation of this guidance document for the validation of gluten methods and the creation of gluten reference materials.

  13. Validation of an ELISA method for the serological diagnosis of canine brucellosis due to Brucella canis.

    Science.gov (United States)

    de Oliveira, Maria Zoraida Daltro; Vale, Vera; Keid, Lara; Freire, Songeli Menezes; Meyer, Roberto; Portela, Ricardo Wagner; Barrouin-Melo, Stella Maria

    2011-06-01

    In the present study, the validation of an enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of canine brucellosis is described. Two different antigenic extracts, obtained by heat or ultrasonic homogenization of microbial antigens from a wild isolate of Brucella canis bacteria, were compared by ELISA and Western blot (WB). A total of 145 canine sera were used to define sensitivity, specificity and accuracy of the ELISA as follows: (1) sera from 34 animals with natural B. canis infection, confirmed by blood culture and PCR, as well as 51 sera samples from healthy dogs with negative results by the agar-gel immunodiffusion (AGID) test for canine brucellosis, were used as the control panel for B. canis infection; and (2) to scrutinize the possibility of cross reactions with other common dog infections in the same geographical area in Brazil, 60 sera samples from dogs harboring known infections by Leptospira sp., Ehrlichia canis, canine distemper virus (CDV), Neospora caninum, Babesia canis and Leishmania chagasi (10 in each group) were included in the study. The ELISA using heat soluble bacterial extract (HE-antigen) as antigen showed the best values of sensitivity (91.18%), specificity (100%) and accuracy (96.47%). In the WB analyses, the HE-antigen showed no cross-reactivity with sera from dogs with different infections, while the B. canis sonicate had various protein bands identified by those sera. The performance of the ELISA standardized with the heat soluble B. canis antigen indicates that this assay can be used as a reliable and practical method to confirm infection by this microorganism, as well as a tool for seroepidemiological studies.

  14. Variants of ELISA in plant virus diagnosis.

    Science.gov (United States)

    Koenig, R; Paul, H L

    1982-10-01

    Variations of enzyme-linked immunosorbent assay (ELISA) were compared with respect to their ability to detect and to differentiate serologically related plant viruses. The broadest range of serologically related viruses was detected by an indirect ELISA on unprecoated plates. Coating the plates with F(ab')2 fragments led to narrowing of the specificity in heterologous reactions of tymo-, tombus- and tobamoviruses in indirect ELISA. With Andean potato latent virus (APLV) heterologous reactions were weaker on plates precoated with F(ab')2 fragments than on those precoated with intact antibodies. Even on plates precoated with F(ab')2 fragments the indirect ELISA detected a broader range of serologically related viruses than the direct double antibody sandwich method. Heterologous reactions in indirect ELISA procedures on plates precoated with either intact antibodies or F(ab')2 fragments were always weaker than homologous reactions independent of the concentration of coating reactants and detecting antibodies. Attempts to differentiate closely related strains of APLV or radish mosaic virus by direct ELISA using F(ab')2 fragments either for coating the plates or after labelling with alkaline phosphatase for detecting the trapped antigens failed. Under suitable conditions, the additional working step usually necessary for indirect ELISA could be avoided by using a short procedure which at low concentrations of detecting antibodies was more sensitive than the conventional procedure.

  15. Preparation of polyclonal antibodies against pneumococcal serotype 1 capsular polysaccharide and development a sandwich ELISA in determination of concentration for type 1 capsular polysaccharide%肺炎球菌1型荚膜多糖多克隆抗体制备与夹心ELISA 法建立及其在多糖浓度测定中的应用

    Institute of Scientific and Technical Information of China (English)

    刘威; 廖红梅; 谢谦; 黄放; 邹莎莎; 马诚; 江山; 崔长法

    2013-01-01

    Objective To prepare polyclonal antibodies against capsular polysaccharide of pneumococcal serotype 1, and use the antibodies to develop a sandwich ELISA in determination of concentration for pneumococcal capsular polysaccharide of serotype 1 in the process of fermentation and purification .Methods High titer serum of anti-capsular polysaccharide se-rotype 1 was obtained after immunizing rabbits by using of inactivated streptococcus pneumoniae cells of serotype 1 for 6 weeks.IgG was purified through affinity chromatography and used as a coating antibody .Polysaccharide samples , alone with in house polysaccharide standard , were then added followed by using biotinylated antibody as a signal antibody .The optimal linear range of the standard curve , specificity, accuracy and precision of the developed method were validated accordingly.Results The antibody titer of rabbit immune serum reached to 1 ∶32, the standard PS1 linear detection range was 1.56 ng/mL to 50 ng/mL, detection limit was 3.13 ng/mL.Standard polysaccharide was mixed with polysaccha-rides from different serotypes or culture media before asssay , the recovery ratio was 102%and 108%respectively .Intra-as-say precision and inter precision of the method were 6.08%and 7.01%.Conclusion High titer rabbit anti-serum against type 1 capsular polysaccharide of streptococcal pneumoniae was prepared .The developed sandwich ELISA method showed high specificity , good accuracy and precision .This method can be used in specific detection of concentration for pneumo-coccal polysaccharide serotype1 .%目的:利用肺炎球菌1型全菌体制备多克隆抗体,并且利用该抗体建立肺炎1型荚膜多糖夹心酶联免疫吸附分析法( Enzyme-linked immunosorbent assay ,ELISA),用于检测发酵和纯化过程中的多糖浓度。方法用灭活的1型肺炎链球菌免疫家兔6周,获得高滴度的抗多糖血清,经过亲和层析纯化,获得高纯度的兔抗肺炎1型多糖

  16. Stochastic sandwich method with low mode substitution for nucleon isovector matrix elements

    CERN Document Server

    Yang, Yi-Bo; Draper, Terrence; Gong, Ming; Liu, Keh-Fei

    2015-01-01

    We introduce a stochastic sandwich method with low-mode substitution to evaluate the connected three-point functions. The isovector matrix elements of the nucleon for the axial-vector coupling $g_A^3$, scalar couplings $g_S^3$ and the quark momentum fraction $\\langle x\\rangle_{u -d}$ are calculated with overlap fermion on 2+1 flavor domain-wall configurations on a $24^3 \\times 64$ lattice at $m_{\\pi} = 330$ MeV with lattice spacing $a = 0.114$ fm.

  17. Sandwich construction

    Science.gov (United States)

    Marshall, A.

    A form of composites known as structural sandwich construction is presented in terms of materials used, design details for solving edging and attachment problems, and charts of design material analysis. Sandwich construction is used in nearly all commercial airliners and helicopters, and military air and space vehicles, and it is shown that this method can stiffen a structure without causing a weight increase. The facing material can be made of 2024 or 7075 aluminum alloy, titanium, or stainless steel, and the core material can be wood or foam. The properties of paper honeycomb and various aluminum alloy honeycombs are presented. Factors pertaining to adhesive materials are discussed, including products given off during cure, bonding pressure, and adaptability. Design requirements and manufacturing specifications are resolved using numerous suggestions.

  18. Comparative Study of Three Different ELISA to Measure the Antibodies Against Infectious Bronchitis Virus in Vaccinated and Unvaccinated Broilers

    Directory of Open Access Journals (Sweden)

    Cardoso TC

    2001-01-01

    Full Text Available Broilers were spray-vaccinated (n=150 with H120 serotype at one-day-old, challenged after 28 days with M41 IBV serotype and after bled at day 28, 34 and 46 after challenged. The respective sera were tested by the indirect ELISA (I-ELISA, sandwich ELISA (S-ELISA, liquid phase blocking ELISA (LPB-ELISA and the standard serum neutralization test (SNT. For this purpose, a total of 300 sera samples, 150 from non vaccinated and 150 from vaccinated broilers were titrated by all the serological methods and the correlation coefficients were determined. The correlation values (r between LPB-ELISA x SNT, S-ELISA x SNT, I-ELISA x SNT found werer = 0.98, r = 0.79, and r = 0.74, respectively. Nevertheless, the r between LPB-ELISA x S-ELISA, LPB-ELISA x I-ELISA and S-ELISA x I-ELISA were r = 0.75, r = 0.69 and r = 0.79. In fact , the I-ELISA and the S-ELISA had almost the same correlation with S-ELISA and LPB-ELISA, in contrast with the I-ELISA and LPB-ELISA. It was concluded that LPB-ELISA showed better sensitivity than I-ELISA and S-ELISA, also after 46 days challenge with the heterologous serotype. However, these two last techniques demonstrated similar specificity when the titers were compared with those obtained in SNT, even though after the heterologous serotype challenge the SNT produced better results. This study demonstrated the close relationship between LPB-ELISA and SNT assays.

  19. Development of a double antibody sandwich ELISA for detection of infectious pancreatic necrosis virus%传染性胰坏死病毒双抗体夹心 ELISA检测方法的建立

    Institute of Scientific and Technical Information of China (English)

    张琳琳; 刘敏; 连科迅; 张英; 蒋烨; 姜艳萍; 崔文; 乔薪瑗; 唐丽杰; 李一经

    2014-01-01

    A double-antibody sandwich ELISA ( DAS-ELISA) was developed using purified polyclonal antibody and mono-clonal antibody ( MAb) against viral infectious pancreatic necrosis virus ( IPNV) VP2 protein as capture and detector anti-body, respectively.The optimized reaction conditions included coating with 100μL/well of purified polyclonal antibody at concentration of 1.28 μg/mL, probing with the working concentration of 1.34 μg/mL IgG from the MAb diluting HRP-conjugated antibody with the proportion of 1∶2 000 and judging with P/N>2 and OD490 nm =0.101 494 as positive criteria. The DAS-ELISA was specific detection of IPNV , but no cross-reaction with IHNV, VHSV, SVCV, HRV and inter-assay coefficient of variability were within 10%.In addition, a total of 41 livers of rainbow trout samples were tested by the DAS-ELISA and PT-PCR, and the coincidence was 97%between DAS-ELISA and PT-PCR.These data demonstrated that the DAS-ELISA was specific and sensitive , and provided a useful tool for diagnosis of IPNV infection .%以纯化的兔抗传染性胰坏死病病毒( IPNV) VP2重组蛋白多克隆抗体为包被抗体,抗IPNV VP2单克隆抗体为检测抗体建立了IPNV双抗体夹心ELISA方法。优化反应条件为:兔抗IPNV VP2重组蛋白多克隆抗体包被浓度为1.28μg/mL,抗IPNV VP2单克隆抗体的工作浓度为1.34μg/mL,酶标二抗稀释比例为1∶2000,以P/N>2,且OD490 nm >0.101494作为阳性判定标准。该方法的重复性变异系数均小于10%,与传染性造血器官坏死病毒(IHNV)、病毒性出血败血症病毒(VHSV)、鲤春病毒血症病毒(SVCV)、轮状病毒(HRV)无交叉反应。对41份虹鳟肝脏样品分别进行双抗体夹心ELISA和RT-PCR检测,结果双抗体夹心法与RT-PCR法检测符合率为97%,表明本实验建立的双抗体夹心ELISA检测方法检测IPNV具有较高的敏感性和特异性,可用于IPNV的病原学检测。

  20. A Comparison of Immuncapture Agglutination and ELISA Methods in Serological Diagnosis of Brucellosis

    Directory of Open Access Journals (Sweden)

    Mehmet Özdemir, Bahadır Feyzioğlu, Muhammed Güzel Kurtoğlu, Metin Doğan, Hatice Türk Dağı, Şerife Yüksekkaya, Recep Keşli, Bülent Baysal

    2011-01-01

    Full Text Available Background: Different serological tests are used in serologic diagnosis of brucellosis. The most widely used of these are Standard Tube Agglutination and Coombs anti-brucella tests. Whereas ELISA Ig M and Ig G tests have been in use for a long time, immuncapture agglutination test has been recently introduced and used in serological diagnosis. The aim of this study was to compare diagnostic values of ELISA Ig M and Ig G and immuncapture agglutination tests with Coombs anti-brucella test.Methods: Sera from 200 patients with presumptive diagnosis of brucellosis were included into the study. Coombs anti-brucella test, ELISA Ig M and Ig G tests and Immuncapture test were investigated in these sera. Then, sensitivity, specificity, negative predictive and positive predictive values were calculated.Results: Sensitivity, specificity, negative predictive and positive predictive values were found to be 90,6 %, 76,3 %, 94,2 %, and 65,9 % respectively for the Immuncapture test, whereas they were found to be 73,7 %, 58,9 %, 84,2 %, and 42,8 % for Ig G and 72,2 %, 67,8 %, 85,2 %, and 48,7 % for Ig M. The Immuncapture test was found to be compatible with ELISA Ig M and Ig G tests but it was statistically incompatible with Coombs anti-brucella test.Conclusions: Immuncapture agglutination test yields similar results to those of Coombs anti-brucella test. This test is a useful test by virtue of the fact that it determines blocking antibodies in the diagnosis and follow-up of brucellosis.

  1. Detection of peste des petits ruminants virus antigen using immunofiltration and antigen-competition ELISA methods.

    Science.gov (United States)

    Raj, G Dhinakar; Rajanathan, T M C; Kumar, C Senthil; Ramathilagam, G; Hiremath, Geetha; Shaila, M S

    2008-06-22

    Peste des petits ruminants (PPR) is one of the most economically important diseases affecting sheep and goats in India. An immunofiltration-based test has been developed using either mono-specific serum/monoclonal antibodies (mAb) prepared against a recombinant truncated nucleocapsid protein of rinderpest virus (RPV) cross-reactive with PPR virus. This method consists of coating ocular swab eluate from suspected animals onto a nitrocellulose membrane housed in a plastic module, which is allowed to react with suitable dilutions of a mAb or a mono-specific polyclonal antibody. The antigen-antibody complex formed on the membrane is then detected by protein A-colloidal gold conjugate, which forms a pink colour. In the immunofiltration test, concordant results were obtained using either PPRV mAb or mono-specific serum. Another test, an antigen-competition ELISA which relies on the competition between plate-coated recombinant truncated 'N' protein of RPV and the PPRV 'N' protein present in ocular swab eluates (sample) for binding to the mono-specific antibody against N protein of RPV (in liquid phase) was developed. The cut-off value for this test was established using reverse transcription polymerase chain reaction (RT-PCR) positive and negative oculo-nasal swab samples. Linear correlation between percent inhibition (PI) values in antigen-competition ELISA and virus infectivity titres was 0.992. Comparison of the immunofiltration test with the antigen-competition ELISA yielded a sensitivity of 80% and specificity of 100%. These two tests can serve as a screening (immunofiltration) and confirmatory (antigen-competition ELISA) test, respectively, in the diagnosis of PPR in sheep or goats.

  2. [Comparison of direct microscopy, culture, ELISA and molecular methods for diagnosis of Entamoeba histolytica].

    Science.gov (United States)

    Tüzemen, Nazmiye Ulkü; Doğan, Nihal

    2014-01-01

    Amebiasis, a parasitic infection caused by Entamoeba histolytica, is one of the most common parasitic infections worldwide. Since it is still an important public health problem in developing countries, rapid differential diagnosis of amebiasis is crucial in terms of treatment. The most frequently used method for laboratory diagnosis is direct microscopy, however more reliable and specific methods are needed in order to differentiate the apathogenic Entamoeba dispar under the microscope. This study was conducted to compare the results of different methods namely, direct microscopy, culture, ELISA and PCR for the detection of E.histolytica in stool samples and to evaluate the performances of those methods. A total of 1049 stool samples collected from pediatric and adult patients who were admitted to hospital with diarrhea complaint between January 2011-March 2013, and randomly selected samples from primary school children, were included in the study. Direct microscopic examination was performed by native-lugol, physiological saline, modified formol-ethyl acetate sedimentation and trichrome staining methods. The stool samples were also inoculated into TYI-S-33 media for axenic cultivation of amoeba. The presence of amebic antigens in the samples were screened by a commercial ELISA kit (TechLab, E.histolytica II, USA). For the molecular diagnosis, a multiplex tandem real-time PCR (MT-PCR) kit (AusDiagnostics Pty Ltd, Australia) was used, after the extraction of DNAs with QIAamp DNA Stool Mini Kit (Qiagen, USA). A total of 354 samples which could be evaluated by all of the methods, were included in the study. Of the 354 stool samples, 84 (23.7%) were found E.histolytica/E.dispar positive by direct microscopy, 61 (17.2%) by trichrome staining, 46 (12.9%) by culture, 31 (8.7%) by ELISA and 9 (2.5%) by MT-PCR. Of direct microscopy positive samples 54.7% (46/84) were also positive with trichrome staining, 39.3% (33/84) with culture, 15.5% (13/84) with ELISA and 7.1% (6

  3. Detection of Epstein- Barr virus infection in lymphoma: ELISA and PCR method

    Directory of Open Access Journals (Sweden)

    Pourakbari B

    2010-02-01

    Full Text Available "n Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:Arial; mso-bidi-theme-font:minor-bidi;} Background: Latent Epstein- Barr virus (EBV genomes are found in the malignant cells of approximately one-third of Hodgkin's lymphoma (HL cases. Detection of EBV viral DNA could potentially be used as a biomarker of disease activity. Our goal was to compare of EBV DNA detection in samples obtained from lymphoma patients versus controls."n"nMethods: One milliliter uncoagulated and 1ml coagulated blood sample for DNA extraction and serum analysis using ELISA for IgG anti EBNA-1 were obtained from 44 lymphoma patients and from 44 normal controls, respectively. EBV genome, EBNA-2, was examined from DNA extracts of paraffin embedded and blood samples using Nested PCR with type specific inner primers."n"nResults: Positive results for ELISA, Blood and biopsy PCR in study group were, 84.1%, 27.3% and 13.6%, respectively. However, these results in control group were 47.7% and 16% for ELISA and Blood PCR assays, respectively. Positive results in ELISA, Blood PCR and Biopsy PCR in Hodgkin and non-Hodgkin patients were found in 21(84%, 6(24%, 4(16% and 16(84.2%, 6(31.6%, 2(10.5% of specimens, respectively. No significant differences in EBV detection were found between these two patient groups (p values for ELISA, Blood PCR

  4. Design Methods of Cell Adhesion Proteins Based on ELISA Usable in-vitro in Gene Therapy

    Directory of Open Access Journals (Sweden)

    E Hosseini

    2016-07-01

    Full Text Available Background & aim: One of the strategies to improve the therapeutic gene is targeting gene therapy. A method which can be considered, is adding code sequences peptide or protein with high tendency to target cells and secreting the therapeutic gene encodes a protein. However, evaluating the effectiveness of such changes in the targeted cell binding protein gene product with the usual therapeutic methods produced in prokaryotic system is directly impossible. The purpose of this study was to evaluate the design methods of cell adhesion proteins based on ELISA usable in-vitro in gene therapy. Methods: In order to target the therapeutic gene Mda-7 by using genetic engineering, peptide coding sequence RGD4C with the tendency to cancerous cell surface integrin were inserted shortly after the artificial signal peptide sequence and the N-terminal coding region of the protein. Then, the modified and unmodified cDNA eukaryotic expression vector pCDNA3.1 were matched. Vectors were transfected in HEK-293 cell line. Then Mda-7 secreted expression levels were measured in cell culture by ELISA. After adjusting the protein concentration of Mda-7 and RGD.Mda-7, in cells transfected media, they were used as a source of protein. Reduce the concentration of these genes was assessed two hours after exposure to the integrin cell lines with HepG2, M21 and lacking integrin Saos-2  were also determined by ELISA. The present study was conducted three times independently.  Data were analyzed using t-test. Results: Statistical analysis of the results suggested that the gene product of the gene product RGD.Mda-7 and Mda-7 to connect to HepG2 cells and M21 were more likely to have integrin. While binding to the cell lines of Saos-2, no significant difference were observed. Conclusions: It seems the present ELISA based method was a suitable strategy for cell attachment assay in gene therapy research.  

  5. Shape and Stress Sensing of Multilayered Composite and Sandwich Structures Using an Inverse Finite Element Method

    Science.gov (United States)

    Cerracchio, Priscilla; Gherlone, Marco; Di Sciuva, Marco; Tessler, Alexander

    2013-01-01

    The marked increase in the use of composite and sandwich material systems in aerospace, civil, and marine structures leads to the need for integrated Structural Health Management systems. A key capability to enable such systems is the real-time reconstruction of structural deformations, stresses, and failure criteria that are inferred from in-situ, discrete-location strain measurements. This technology is commonly referred to as shape- and stress-sensing. Presented herein is a computationally efficient shape- and stress-sensing methodology that is ideally suited for applications to laminated composite and sandwich structures. The new approach employs the inverse Finite Element Method (iFEM) as a general framework and the Refined Zigzag Theory (RZT) as the underlying plate theory. A three-node inverse plate finite element is formulated. The element formulation enables robust and efficient modeling of plate structures instrumented with strain sensors that have arbitrary positions. The methodology leads to a set of linear algebraic equations that are solved efficiently for the unknown nodal displacements. These displacements are then used at the finite element level to compute full-field strains, stresses, and failure criteria that are in turn used to assess structural integrity. Numerical results for multilayered, highly heterogeneous laminates demonstrate the unique capability of this new formulation for shape- and stress-sensing.

  6. Determination of aflatoxins in nuts of Tabriz confectionaries by ELISA and HPLC methods

    Directory of Open Access Journals (Sweden)

    Siahi Shadbad Mohammad Reza

    2012-06-01

    Full Text Available Purpose: Aflatoxins (AFs are a group of mycotoxins and secondary metabolites of various species of Aspergillus. There are various forms of aflatoxins including B1, B2, G1, G2, M1 and M2 types. Aflatoxins cause important health problems and have high potential effect on liver cancer. Therefore, numerous investigations have been conducted during last three decades. The aim of this work is to determine the contamination levels of nuts used by the confectionaries in Tabriz. Methods: A total of 142 samples including 35 almond , 26 walnut, 4 seeds of apricot, 6 sunflower seeds kernel, 6 sesame seed, 6 peanuts , 32 pistachio,13 hazelnuts and 14 cashews samples were collected from Tabriz confectionaries. The ELISA method was employed for the screening of total aflatoxins. Results: In 13 cases (28.1% of pistachios, 5.1% of walnuts and 7.1% of cashews contamination rate of higher than 15 ppb were observed. The HPLC method was applied for the confirmation of ELISA results. Aflatoxin B1 was the highest detected AFs. Conclusion: The overall results of the tested samples indicated that the rate of contamination of pistachios is higher than the other tested samples.

  7. A New Method with Sandwiched Composite Films for Encapsulating Flexible OLEDs

    Institute of Scientific and Technical Information of China (English)

    LI Yang; WANG Li-Duo; DUAN Lian; QIU Yong

    2005-01-01

    @@ We introduce a novel method for sandwiched-composite-film encapsulation that successfully extends the lifetime of flexible organic light-emitting diodes (FOLEDs). The encapsulation layers include two parts: one is a thin multilayer barrier coating, which is made up of two applications of alternating layers composed of a polymer layer (consisting of UV capable resins) and a ceramic layer (consisting of titanium nitride with excellent barrier performance), and the other is a thick polymer film of approximately 70μm in thickness fabricated by a doctor blade onto the thin encapsulation film described above. FOLEDs encapsulated by this novel method have a longer lifetime, and this lifetime is 74 times as much as the lifetime of unencapsulated ones.

  8. The validity of some haematological and ELISA methods for the diagnosis of canine heartworm disease.

    Science.gov (United States)

    Martini, M; Capelli, G; Poglayen, G; Bertotti, F; Turilli, C

    1996-01-01

    Examinations for heartworm (Dirofilaria immitis) were performed on 175 impounded dogs from a hyperendemic area of the Po Valley (Italy). Each blood sample was used with five haematological diagnostic methods (filtration, direct smear, modified Knott, clotted blood and capillary tube) and three commercial ELISA kits (PetChek, Diasystems, Uni-Tec). The results were compared with the true infection status obtained from post-mortem examination of the heart, pulmonary arteries, thoracic venae cavae and lungs. The prevalence of the infection by adult worms at necropsy was 63%. The sensitivity of the tests ranged from 60% (capillary tube) to 81% (Diasystems) and the specificity from 88% (filtration) to 98% (PetChek). The results of all the tests differed significantly (p < 0.01) from those obtained at necropsy. The sensitivity of the tests was also assessed with respect to the differing numbers of worms in the hosts. A positive correlation between the worm burden and the sensitivity was observed in all the tests. It is apparent that the ELISA methods were better able to detect cases with a low number of worms than the haematological tests.

  9. [Contamination level of aflatoxin B1 in lotus seeds rapid screening by indirect competitive ELISA method].

    Science.gov (United States)

    Chu, Xian-feng; Dou, Xiao-wen; Kong, Wei-jun; Yang, Mei-hua; Zhao, Chong; Zhao, Ming; Ouyang, Zhen

    2015-02-01

    A simple and cost-effective indirect competitive enzyme-linked immune sorbent assay (ic-ELISA) was developed to rapidly screen the content of aflatoxin B1 (AFB1) in lotus seeds, and the results were confirmed by ultra-fast liquid chromatography-tandem mass spectrometry( UFLC-MS/MS). Matrix-matched calibration expressed a good linearity ranging from 0. 171 to 7. 25 µg · L(-1) for AFB, with R2 > 0.978. The medium inhibitory concentration( IC50 ) for AFB1 was 1.29 µg · L(-1), the recovery for AFB1 was 74.73% to 126.9% with RSD lotus seeds samples and the results indicated that the contents of AFB, in samples 1-15 were in the range of 1. 19- 115. 3 µg · kg(-1) and in 40% of the samples exceeded the legal limit(5 µg · kg(-1)), while the contamination rate of AFB, in samples 16-20 was 40%. Pearson correlation coefficient(r) reached 0.997 for AFB1 content in the samples detected by ic-ELSIA and UFLC-MS/MS methods. The results proved that the developed ic-ELISA method is simple, sensitive and reliable, and can be used for rapid and high-throughput screening of AFB1 in lotus seeds

  10. Adjuvant chemo-radiotherapy in the "sandwich" method for high risk endometrial cancer--a review of literature.

    Science.gov (United States)

    Bie, Yachun; Zhang, Zhenyu; Wang, Xiaolan

    2015-06-24

    Endometrial cancer is a common female malignancy. Patients with high-risk endometrial cancer have relatively high incidence of metastasis and recurrence. Despite complete resection, patients with stage III or IV are at high risk of local or distant recurrence. Systemic adjuvant treatment includes chemotherapy and radiotherapy. But the optimal scheduling is not known. Recently proposed sequential chemo-radiotherapy as sandwich therapy for high risk endometrial cancer have yielded encouraging results. This article is to review the adjuvant chemo-radiotherapy in the "sandwich" method for high risk endometrial cancer to help clinicians identify the most effective adjuvant treatment for patients with high risks of it. We used MEDLINE, EMBASE, Cochrane Library and CBM databases to search the literature. A systematic review was made. And most data showed "sandwich" therapy is feasible, efficacious, well-tolerated and resulted in excellent long-term progression free and overall survival in the setting of advanced endometrial cancer. Randomized trials are necessary to compare chemo-radio therapy given in the "sandwich" fashion to other means of sequencing these treatment modalities. It is also necessary to define which population is best suited for "sandwich" adjuvant therapy.

  11. Detection of Vibrio anguillarum and Its Virulent Metalloprotease Using the Method of ELISA

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    A method detecting pathogenic Vibrio anguillarum and its virulent metalloprotease is reported in this paper. The metalloprotease is isolated from extracellular product (ECP) of V.anguillarum by the cellophane plate technique and purified by gel filtration and ion-exchange chromatography. Anti-sera are prepared by injecting V.anguillarum cells and metalloprotease into the rabbits. Slide Agglutination Assay is used to detect V.anguillarum in the infection experiment and Enzyme Linked Immunosorbent Assay (ELISA) is carried out to detect concentration of metalloprotease. The result shows that bacterium strain M3 is able to diffuse into the viscera of infected fish through the blood circulating system 10 hours after intramuscular infection, and ELASA is a sensitive method to detect the metalloprotease with detectable amount of 7.8ng. The aim of this study is to establish a sensitive and specific method to observe the infection of V.anguillarum in the host.

  12. A Sandwich Method Using Kapandji Intrafocal Pinning to Facilitate Palmar Plating of Displaced Distal Radius Fractures.

    Science.gov (United States)

    Huang, Hui-Kuang; Huang, Yi-Chao; Wang, Jung-Pan; Chang, Ming-Chau

    2017-09-01

    The use of palmar plating in the treatment of dorsally displaced distal radius fracture is very common, and can result in a good functional outcome. We present an easy "sandwich method" to reduce the fracture and to facilitate osteosynthesis. Firstly, the dorsal Kapandji intrafocal wire was applied to achieve the reduction of the dorsally displaced fracture and provide a volar-directed force. Then the intra-articular fractures were reduced or the metaphyseal defect is stuffed with bone graft if necessary. Finally, the anatomical plate is used to buttress and push back the distal fragment to complete the osteosynthesis. The postoperative radiographic parameters, comparing with the contralateral noninjured side, could achieve similar radial height, radial inclination, volar tilt, and ulnar variance without significant difference.

  13. Chromatofocusing combined with the ELISA technique. A sensitive method for the analysis of immune complexes.

    Science.gov (United States)

    Kneba, M; Krieger, G; Kehl, A; Bause, I; Nagel, G A

    1983-07-15

    A sensitive method which permits analysis of IgG containing circulating immune complexes without detailed knowledge of the nature of the antigens and the specificity of the antibodies involved is described. Soluble BSA: anti-BSA were used as model immune complexes and isolated from serum. The procedure involves the use of gel chromatography for the separation of the high molecular weight fraction containing the immune complexes as measured by binding to 125I-labeled Clq, followed by absorption of the immune complex fraction to immobilized protein A-Sepharose CL-4B. After desorption from protein A-Sepharose the complexes were dissociated and separated into free antigen and antibody by chromatofocusing in the presence of urea. The isolated free antigen and antibody retained their immunological activity as shown by immunodiffusion, binding after their recombination to 125I-labeled Clq, and by recombining antigen and antibody with much enhanced sensitivity using a microplate ELISA system. By means of the ELISA recombination technique it is possible to analyze less than 1 microgram of BSA:anti-BSA model complexes. Application of this technique may provide more information about the nature of immune complex like material associated with diseases.

  14. An ELISA Based Binding and Competition Method to Rapidly Determine Ligand-receptor Interactions.

    Science.gov (United States)

    Syedbasha, Mohameedyaseen; Linnik, Janina; Santer, Deanna; O'Shea, Daire; Barakat, Khaled; Joyce, Michael; Khanna, Nina; Tyrrell, D Lorne; Houghton, Michael; Egli, Adrian

    2016-01-01

    A comprehensive understanding of signaling pathways requires detailed knowledge regarding ligand-receptor interaction. This article describes two fast and reliable point-by-point protocols of enzyme-linked immunosorbent assays (ELISAs) for the investigation of ligand-receptor interactions: the direct ligand-receptor interaction assay (LRA) and the competition LRA. As a case study, the ELISA based analysis of the interaction between different lambda interferons (IFNLs) and the alpha subunit of their receptor (IL28RA) is presented: the direct LRA is used for the determination of dissociation constants (KD values) between receptor and IFN ligands, and the competition LRA for the determination of the inhibitory capacity of an oligopeptide, which was designed to compete with the IFNLs at their receptor binding site. Analytical steps to estimate KD and half maximal inhibitory concentration (IC50) values are described. Finally, the discussion highlights advantages and disadvantages of the presented method and how the results enable a better molecular understanding of ligand-receptor interactions.

  15. An ELISA method using serum derived HDAg for the sorological detection of HDV antigens and antibodies

    Directory of Open Access Journals (Sweden)

    Celso Granato

    1987-12-01

    Full Text Available One of the main difficulties related to the detection of the Hepatitis Delta Virus (HDV antigen and antibody has been the source of the needed HD antigen since HDV containing human and animal livers are very difficult to obtain and since yield is low. This fact prompted us to try to use the serum of patients in the acute phase of HDV infection as a source of HDAg and turn to enzyme immunoassays (EIA instead of RIA for the sake of easiness and economy in the amount of HDAg needed. The antigen for EIA was obtained from patients during the acute phase of HDV infection and the antibody from patients who have been carriers for many years. For the detection of the antigen, a sandwich type method was employed, whereas for the antibody a competition assay was developed. In order to assess the relative specificity and sensibility of the test, the antibody assay was compared to a commercial RIA (C. RIA, Abbott and to a non-commercial RIA (NC RIA. Forty-two sera were tested by the two methods and only in two cases discrepant results were obtained. Its is concluded that: 1 sera from patients in the acute and chronic phases of HDV infection can be used as source of both antigen and antibody, for immunoassays; 2 EIA and RIA have comparable relative specificity and sensibility and 3 EIA is easier to perform, cheaper, non-hazardous, has a longer shelf-life and saves scarce HDAg.

  16. Validation study of a rapid ELISA for detection of aflatoxin in corn. Performance Tested Method 050901.

    Science.gov (United States)

    Lupo, Anthony; Roebuck, Chris; Dutcher, Monica; Kennedy, Justina; Abouzied, Mohamed

    2010-01-01

    Neogen Corp. developed the Veratox aflatoxin test kit for the detection of total aflatoxin. The purpose of this study was to validate the method under the requirements of the AOAC Research Institute Performance Tested Methods (PTM) program. There are several AOAC Official Methods for total aflatoxin detection in corn (994.08, 990.33, 979.18, 993.17, 990.32, 993.16, 991.31, and 990.74), varying between rapid and analytical-based methods and one rapid method that has been performance tested by the AOAC Research Institute (PTM 030701). However, the widely used reference method is AOAC Official Method 994.08, which is an HPLC method and is referred to as the reference method in this paper. Although considered the reference method, the HPLC procedure is complicated and requires the investment of both expensive equipment and a highly skilled technician. A rapid (e.g., ELISA) test kit to be validated by the AOAC Research Institute is needed.

  17. Validation of an ELISA method for the serological diagnosis of canine brucellosis due to Brucella canis

    OpenAIRE

    Oliveira, Maria Zoraida Daltro de; Vale, Vera; Keid, Lara; Freire, Songeli Menezes; Nascimento, Roberto José Meyer; Portela, Ricardo Wagner Dias; Melo, Stella Maria Barrouin

    2011-01-01

    Acesso restrito: Texto completo. p. 425-431. In the present study, the validation of an enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of canine brucellosis is described. Two different antigenic extracts, obtained by heat or ultrasonic homogenization of microbial antigens from a wild isolate of Brucella canis bacteria, were compared by ELISA and Western blot (WB). A total of 145 canine sera were used to define sensitivity, specificity and accuracy of the ELISA as f...

  18. Concentrations of haptoglobin in bovine plasma determined by ELISA or a colorimetric method based on peroxidase activity.

    Science.gov (United States)

    Cooke, R F; Arthington, J D

    2013-06-01

    The objective was to compare different procedures for determination of haptoglobin in bovine plasma. Nine Angus steers were vaccinated against Mannheimia haemolytica to stimulate an acute-phase response. Blood samples were collected immediately prior to vaccination (day 0), and on days 1, 3, 5, 7 and 10. Plasma samples were frozen in duplicates at -80 °C. One set of the duplicates was analysed for haptoglobin concentrations using a commercial ELISA kit. A day effect was detected (p peroxidase activity (CPPA) with results expressed as optical density. Further, based on the ELISA results, the plasma sample with the greatest haptoglobin concentration was also serially diluted into a plasma sample with negligible haptoglobin concentration from the same steer (1:1 through 1:1024 dilution). These dilutions were used within the CPPA method to generate a standard curve and estimate plasma haptoglobin concentrations (CPPA + STD). A linear standard curve was generated (r(2)  = 0.99). A day effect similar to the ELISA method was detected for the CPPA and CPPA + STD methods (p ELISA methods were positively correlated (r = 0.97; p ELISA method. In conclusion, assessing concentrations of haptoglobin in bovine plasma using the CPPA and CPPA + STD methods generate highly correlated or similar results, respectively, compared to ELISA. Therefore, the CPPA + STD and CPPA methods can be used as a less expensive alternative to ELISA to determine concentrations or monitor changes in plasma haptoglobin in bovine samples.

  19. Sensitive detection of Escherichia coli O157:H7 based on cascade signal amplification in ELISA.

    Science.gov (United States)

    Shan, Shan; Liu, Daofeng; Guo, Qi; Wu, Songsong; Chen, Rui; Luo, Kai; Hu, Liming; Xiong, Yonghua; Lai, Weihua

    2016-09-01

    In this study, cascade signal amplification in ELISA involving double-antibody sandwich ELISA and indirectly competitive ELISA was established to sensitively detect Escherichia coli O157:H7. In the double-antibody sandwich ELISA, a complex was formed comprising anti-E. coli O157:H7 polyclonal antibody, E. coli O157:H7, biotinylated anti-E. coli O157:H7 monoclonal antibody, streptavidin, and biotinylated β-lactamase. Penicillin solution was then added into the ELISA well and hydrolyzed by β-lactamase. Afterward, the penicillin solution was transferred to indirectly competitive ELISA. The concentration of penicillin can be sensitively detected in indirectly competitive ELISA. In the cascade signal amplification system, increasing the amount of added E. coli O157:H7 resulted in more β-lactamase and less penicillin. The detection sensitivity of E. coli O157:H7, which was 20cfu/mL with the cascade signal amplification in ELISA, was 1,000-fold higher than that of traditional ELISA. Furthermore, the novel method can be used to detect E. coli O157:H7 in milk (2cfu/g). Therefore, this new signaling strategy will facilitate analyses of highly sensitive foodborne pathogens.

  20. An ELISA for the quantitation of von Willebrand Factor

    DEFF Research Database (Denmark)

    Vinholt, Pernille Just; Overgaard, Martin; Diederichsen, Axel Cosmus Pyndt;

    2013-01-01

    with and without documented coronary calcification (total n=118). RESULTS AND CONCLUSIONS: The assay detected VWF:OPG complexes in human plasma, while no significant signal was observed when testing solutions containing VWF or recombinant OPG alone. Importantly, the ELISA assay was able to detect in vitro formed...... for measurement of von Willebrand factor-osteoprotegerin complex (VWF:OPG) in human plasma. Furthermore, the significance of VWF:OPG complex as a marker of cardiovascular disease (CVD) was evaluated. PATIENTS/METHODS: A sandwich ELISA for quantification of VWF:OPG was developed using a polyclonal rabbit anti...

  1. Validation of a digital photographic method for assessment of dietary quality of school lunch sandwiches brought from home

    Directory of Open Access Journals (Sweden)

    Marianne S. Sabinsky

    2013-07-01

    Full Text Available Background: It is a challenge to assess children's dietary intake. The digital photographic method (DPM may be an objective method that can overcome some of these challenges. Objective: The aim of this study was to evaluate the validity and reliability of a DPM to assess the quality of dietary intake from school lunch sandwiches brought from home among children aged 7–13 years. Design: School lunch sandwiches (n=191 were prepared to represent randomly selected school lunch sandwiches from a large database. All components were weighed to provide an objective measure of the composition. The lunches were photographed using a standardised DPM. From the digital images, the dietary components were estimated by a trained image analyst using weights or household measures and the dietary quality was assessed using a validated Meal Index of Dietary Quality (Meal IQ. The dietary components and the Meal IQ obtained from the digital images were validated against the objective weighed foods of the school lunch sandwiches. To determine interrater reliability, the digital images were evaluated by a second image analyst. Results: Correlation coefficients between the DPM and the weighed foods ranged from 0.89 to 0.97. The proportion of meals classified in the same or an adjacent quartile ranged from 98% (starch to 100% (fruits, vegetables, fish, whole grain, and Meal IQ. There was no statistical difference between fish, fat, starch, whole grains, and Meal IQ using the two methods. Differences were found for fruits and vegetables; Bland–Altman analyses showed a tendency to underestimate high amounts of these variables using the DPM. For interrater reliability, kappa statistics ranged from 0.59 to 0.82 across the dietary components and Meal IQ. Conclusions: The standardised DPM is a valid and reliable method for assessing the dietary quality of school lunch sandwiches brought from home.

  2. Development and use of specific ELISA methods for quantifying the biological activity of bevacizumab, cetuximab and trastuzumab in stability studies.

    Science.gov (United States)

    Suárez, Inmaculada; Salmerón-García, Antonio; Cabeza, José; Capitán-Vallvey, Luis Fermín; Navas, Natalia

    2016-10-01

    Bevacizumab (BVZ), cetuximab (CTX) and trastuzumab (TTZ) are monoclonal antibodies (mAbs) used worldwide for the treatment of several widespread kinds of cancer. They are marketed as medicines under their respective tradenames: Avastin(®), Erbitux(®) and Herceptin(®). The aim of this research was to develop in-house specific enzyme-linked immunosorbent assays (ELISA) to assess the long-term stability of these three mabs. These assays assess the biological functionality of the mAbs by quantifying their biological activity. For this purpose, we developed an indirect ELISA procedure whereby the specific antigens against which the mAbs are directed are used as specific "capturing" antibodies on the ELISA plates. We therefore used vascular endothelial growth factor (VEGF) in the ELISA for BVZ; human epidermal growth factor receptor (hEGFR) in the ELISA for CTX and human receptor HER2 (hHER2) in the ELISA for TTZ. After the mAbs had attached to their antigen, we used an anti-human IgG (whole molecule) peroxidase-conjugate and o-phenylenediaminedihydrochloride substrate. The reaction was stopped using sulphuric acid and absorbance was recorded at a wavelength of 450nm. The three ELISA methods were validated in terms of calibration models, range of the assay, limits of detection and quantitation, intra and interday precision and accuracy, and specificity by cross reactions. Forced degradation studies were also conducted on the medicines, providing useful information. Finally, the proposed ELISA were successfully used in a long-term stability study to quantify the remaining biological activity in medicines that had been opened and then stored under two different storage conditions, i.e. refrigerated at 4°C and frozen at -20°C. Results indicated that BVZ (Avastin(®)) is the most stable of the three in terms of its biological functionality.

  3. A Simple and Specific Noncompetitive ELISA Method for HT-2 Toxin Detection

    Science.gov (United States)

    Arola, Henri O.; Tullila, Antti; Nathanail, Alexis V.; Nevanen, Tarja K.

    2017-01-01

    We developed an HT-2 toxin-specific simple ELISA format with a positive read-out. The assay is based on an anti-immune complex (IC) scFv antibody fragment, which is genetically fused with alkaline phosphatase (AP). The anti-IC antibody specifically recognizes the IC between a primary anti-HT-2 toxin Fab fragment and an HT-2 toxin molecule. In the IC ELISA format, the sample is added together with the scFv-AP antibody to the ELISA plate coated with the primary antibody. After 15 min of incubation and a washing step, the ELISA response is read. A competitive ELISA including only the primary antibody recognizes both HT-2 and T-2 toxins. The anti-IC antibody makes the assay specific for HT-2 toxin, and the IC ELISA is over 10 times more sensitive compared to the competitive assay. Three different naturally contaminated matrices: wheat, barley and oats, were used to evaluate the assay performance with real samples. The corresponding limits of detection were 0.3 ng/mL (13 µg/kg), 0.1 ng/mL (4 µg/kg) and 0.3 ng/mL (16 µg/kg), respectively. The IC ELISA can be used for screening HT-2 toxin specifically and in relevant concentration ranges from all three tested grain matrices. PMID:28425967

  4. A Simple and Specific Noncompetitive ELISA Method for HT-2 Toxin Detection

    Directory of Open Access Journals (Sweden)

    Henri O. Arola

    2017-04-01

    Full Text Available We developed an HT-2 toxin-specific simple ELISA format with a positive read-out. The assay is based on an anti-immune complex (IC scFv antibody fragment, which is genetically fused with alkaline phosphatase (AP. The anti-IC antibody specifically recognizes the IC between a primary anti-HT-2 toxin Fab fragment and an HT-2 toxin molecule. In the IC ELISA format, the sample is added together with the scFv-AP antibody to the ELISA plate coated with the primary antibody. After 15 min of incubation and a washing step, the ELISA response is read. A competitive ELISA including only the primary antibody recognizes both HT-2 and T-2 toxins. The anti-IC antibody makes the assay specific for HT-2 toxin, and the IC ELISA is over 10 times more sensitive compared to the competitive assay. Three different naturally contaminated matrices: wheat, barley and oats, were used to evaluate the assay performance with real samples. The corresponding limits of detection were 0.3 ng/mL (13 µg/kg, 0.1 ng/mL (4 µg/kg and 0.3 ng/mL (16 µg/kg, respectively. The IC ELISA can be used for screening HT-2 toxin specifically and in relevant concentration ranges from all three tested grain matrices.

  5. DPI-ELISA: a fast and versatile method to specify the binding of plant transcription factors to DNA in vitro

    Directory of Open Access Journals (Sweden)

    Chaban Christina

    2010-11-01

    Full Text Available Abstract Background About 10% of all genes in eukaryote genomes are predicted to encode transcription factors. The specific binding of transcription factors to short DNA-motifs influences the expression of neighbouring genes. However, little is known about the DNA-protein interaction itself. To date there are only a few suitable methods to characterise DNA-protein-interactions, among which the EMSA is the method most frequently used in laboratories. Besides EMSA, several protocols describe the effective use of an ELISA-based transcription factor binding assay e.g. for the analysis of human NFκB binding to specific DNA sequences. Results We provide a unified protocol for this type of ELISA analysis, termed DNA-Protein-Interaction (DPI-ELISA. Qualitative analyses with His-epitope tagged plant transcription factors expressed in E. coli revealed that EMSA and DPI-ELISA result in comparable and reproducible data. The binding of AtbZIP63 to the C-box and AtWRKY11 to the W2-box could be reproduced and validated by both methods. We next examined the physical binding of the C-terminal DNA-binding domains of AtWRKY33, AtWRKY50 and AtWRKY75 to the W2-box. Although the DNA-binding domain is highly conserved among the WRKY proteins tested, the use of the DPI-ELISA discloses differences in W2-box binding properties between these proteins. In addition to these well-studied transcription factor families, we applied our protocol to AtBPC2, a member of the so far uncharacterised plant specific Basic Pentacysteine transcription factor family. We could demonstrate binding to GA/TC-dinucleotide repeat motifs by our DPI-ELISA protocol. Different buffers and reaction conditions were examined. Conclusions We successfully applied our DPI-ELISA protocol to investigate the DNA-binding specificities of three different classes of transcription factors from Arabidopsis thaliana. However, the analysis of the binding affinity of any DNA-binding protein to any given DNA

  6. [Development of immunoenzyme methods for detecting antibodies to Aujeszky's disease virus gB glycoprotein in swine serum].

    Science.gov (United States)

    Morenkov, O S

    2000-01-01

    Four ELISA methods have been developed for detecting antibodies to Aujeszky's disease virus (ADV) glycoprotein gB. Indirect ELISA is based on affinity-purified gB (affi-gB-ELISA); three blocking ELISAs: indirect blocking ELISA (lbgB-ELISA), direct blocking ELISA (db-gB-ELISA), and two-site "sandwich" ELISA (sb-gB-ELISA) are based on monoclonal antibodies to conservative immunodominant epitopes of gB. The specificities and sensitivities of ELISAs were compared with each other and with indirect ELISA based on purified ADV virions (vir-ELISA). Affi-gB ELISA, db-gB-ELISA, and sb-gB-ELISA possess 100% sensitivity, ib-gB-ELISA 98% sensitivity, and vir-ELISA 93% sensitivity. Affi-gB ELISA, ib-gB-ELISA, db-gB-ELISA, and sb-gB-ELISA possess 100% specificity and vir-ELISA 92% specificity. The efficiency of detection of ADV-specific antibodies by affi-gB ELISA, db-gB-ELISA, and sb-gB-ELISA was comparable to that of analogous commercial test. Since db-gB-ELISA is easier to perform than affi-gB-ELISA or sb-gB-ELISA, it is concluded to be the most appropriate test for detecting pigs infected with ADV among non-vaccinated animals.

  7. Finite element method calculations of GMI in thin films and sandwiched structures: Size and edge effects

    Energy Technology Data Exchange (ETDEWEB)

    Garcia-Arribas, A. [Departamento de Electricidad y Electronica, Universidad del Pais Vasco, Apartado 644, 48080 Bilbao (Spain)], E-mail: alf@we.lc.ehu.es; Barandiaran, J.M.; Cos, D. de [Departamento de Electricidad y Electronica, Universidad del Pais Vasco, Apartado 644, 48080 Bilbao (Spain)

    2008-07-15

    The impedance values of magnetic thin films and magnetic/conductor/magnetic sandwiched structures with different widths are computed using the finite element method (FEM). The giant magneto-impedance (GMI) is calculated from the difference of the impedance values obtained with high and low permeability of the magnetic material. The results depend considerably on the width of the sample, demonstrating that edge effects are decisive for the GMI performance. It is shown that, besides the usual skin effect that is responsible for GMI, an 'unexpected' increase of the current density takes place at the lateral edge of the sample. In magnetic thin films this effect is dominant when the permeability is low. In the trilayers, it is combined with the lack of shielding of the central conductor at the edge. The resulting effects on GMI are shown to be large for both kinds of samples. The conclusions of this study are of great importance for the successful design of miniaturized GMI devices.

  8. Development and application of a double-antigen sandwich enzyme-linked immunosorbent assay for detection of antibodies to porcine circovirus 2.

    Science.gov (United States)

    Ge, Meng; Luo, Wei; Jiang, Daliang; Li, Runcheng; Zhao, Wenwei; Chen, Guoliang; Yang, Xingdong; Yu, Xinglong

    2012-09-01

    A double-antigen sandwich enzyme-linked immunosorbent assay (ELISA) is described for detection of porcine circovirus 2 (PCV2) antibodies using the well-characterized recombinant PCV2 capsid protein. In a comparative test of 394 pig sera against an indirect immunofluorescence (IIF) test and a commercial ELISA kit (also based on the recombinant PCV2 capsid protein), the results showed that the diagnostic sensitivity, specificity, and accuracy of the assay were, respectively, 90.61, 94.02, and 91.62% compared with IIF and 94.38, 95.28, and 94.67% compared with the commercial ELISA kit. Assay of 12 PCV-free pigs over a 5-week period produced only PCV2-negative titers by all 3 methods. These results and the seroprofiles of 4 pig farms obtained by both the commercial ELISA kit and the double-antigen sandwich ELISA indicate that the sandwich ELISA is a reliable method for detection of antibodies to PCV2. Additionally, the method described here permits the use of undiluted test serum samples simultaneously loaded with horseradish peroxidase (HRP)-conjugated antigen into the test well, and the complete test procedure can be performed in less than 90 min. This double-antigen sandwich ELISA should be a useful tool to aid swine industry professionals in deciding the intervention strategies for the control of PCV2-associated diseases.

  9. A short history, principles, and types of ELISA, and our laboratory experience with peptide/protein analyses using ELISA.

    Science.gov (United States)

    Aydin, Suleyman

    2015-10-01

    Playing a critical role in the metabolic homeostasis of living systems, the circulating concentrations of peptides/proteins are influenced by a variety of patho-physiological events. These peptide/protein concentrations in biological fluids are measured using various methods, the most common of which is enzymatic immunoassay EIA/ELISA and which guide the clinicians in diagnosing and monitoring diseases that inflict biological systems. All the techniques where enzymes are employed to show antigen-antibody reactions are generally referred to as enzymatic immunoassay EIA/ELISA method. Since the basic principles of EIA and ELISA are the same. The main objective of this review is to present an overview of the historical journey that had led to the invention of EIA/ELISA, an indispensible method for medical and research laboratories, types of ELISA developed after its invention [direct (the first ELISA method invented), indirect, sandwich and competitive methods], problems encountered during peptide/protein analyses (pre-analytical, analytical and post-analytical), rules to be followed to prevent these problems, and our laboratory experience of more than 15 years. Copyright © 2015 Elsevier Inc. All rights reserved.

  10. Semi-in situ atomic force microscopy imaging of intracellular neurofilaments under physiological conditions through the 'sandwich' method.

    Science.gov (United States)

    Sato, Fumiya; Asakawa, Hitoshi; Fukuma, Takeshi; Terada, Sumio

    2016-08-01

    Neurofilaments are intermediate filament proteins specific for neurons and characterized by formation of biochemically stable, obligate heteropolymers in vivo While purified or reassembled neurofilaments have been subjected to morphological analyses by electron microscopy and atomic force microscopy, there has been a need for direct imaging of cytoplasmic genuine intermediate filaments with minimal risk of artefactualization. In this study, we applied the modified 'cells on glass sandwich' method to exteriorize intracellular neurofilaments, reducing the risk of causing artefacts through sample preparation. SW13vim(-) cells were double transduced with neurofilament medium polypeptide (NF-M) and alpha-internexin (α-inx). Cultured cells were covered with a cationized coverslip after prestabilization with tannic acid to form a sandwich and then split into two. After confirming that neurofilaments could be deposited on ventral plasma membranes exposed via unroofing, we performed atomic force microscopy imaging semi-in situ in aqueous solution. The observed thin filaments, considered to retain native structures of the neurofilaments, exhibited an approximate periodicity of 50-60 nm along their length. Their structural property appeared to reflect the morphology formed by their constituents, i.e. NF-M and α-inx. The success of semi-in situ atomic force microscopy of exposed bona fide assembled neurofilaments through separating the sandwich suggests that it can be an effective and alternative method for investigating cytoplasmic intermediate filaments under physiological conditions by atomic force microscopy.

  11. [Levels of Ochratoxin A and total Aflatoxins in Panamanian exportation coffee by an ELISA Method].

    Science.gov (United States)

    Franco, Heriberto; Vega, Aracelly; Reyes, Stephany; De Léon, Javier; Bonilla, Alexis

    2014-03-01

    A study about processing conditions of exportation coffee in 15 benefits located in Chiriqui, western region of Panama, was conducted. In addition, 21 samples of processed coffee (green beans), from the benefits, were analyzed. The samples were microbiologically tested in order to quantify total aflatoxins (B1, B2, G1 and G2) and Ochratoxin A (OTA), using the immunoaffinity ELISA method. A detection limit of 0.017 ng/mL, was determined for Ochratoxin A, which is equivalent to a concentration of 0.829 µg/kg, and a detection limit of 0.027 ng/mL, for total aflatoxins, which is equivalent to a concentration of 1.350 µg/kg. It was found that four (19%) out of the 21 samples were positive to the presence of Ochratoxin A and three (14%) to the presence of total aflatoxins. Samples showed levels of Ochratoxin A in the range 4.90 - 37.73 µg/kg; only three of them exceeded the maximum limit allowed by the European Union, for the concentration of Ochratoxin, which is of 5.0 µg/kg. Total aflatoxins were found in the range 1.51 - 1.93 µg/kg, below 10 µg/kg which is the maximum limit allowed for coffee by the European Union. The results indicate that the processing of coffee produced in Panama successfully meets international standards for postharvest handling, which leads to a low incidence ofmycotoxins and very low levels ofmycotoxin-producing fungi.

  12. Sandwiched composites in aerospace engineering

    OpenAIRE

    Nunes, J. P.; Silva,J.F.

    2016-01-01

    This chapter considers sandwiched composites used in aerospace applications. Typical sandwich composites consist of two thin, stiff, high-strength facing skins separated by a thick and light core. New developments in the type of face and core materials, production methods and joining and repair techniques are discussed in this chapter. It also discusses various properties as well as their main design methods for existing and future applications of sandwiched composites.

  13. Development of a sulfonamide ELISA and its comparison with LC-MS/MS method

    Science.gov (United States)

    Sulfonamides are common chemotherapeutic agents used in veterinary and human medicines to treat bacterial and protozoa infections. Because of their widespread usage sulfonamides, such as sulfamethoxazole, are some of the most prevalent pharmaceuticals found in waterways. A sulfonamide ELISA has be...

  14. Predictions of thermal buckling strengths of hypersonic aircraft sandwich panels using minimum potential energy and finite element methods

    Science.gov (United States)

    Ko, William L.

    1995-01-01

    Thermal buckling characteristics of hypersonic aircraft sandwich panels of various aspect ratios were investigated. The panel is fastened at its four edges to the substructures under four different edge conditions and is subjected to uniform temperature loading. Minimum potential energy theory and finite element methods were used to calculate the panel buckling temperatures. The two methods gave fairly close buckling temperatures. However, the finite element method gave slightly lower buckling temperatures than those given by the minimum potential energy theory. The reasons for this slight discrepancy in eigensolutions are discussed in detail. In addition, the effect of eigenshifting on the eigenvalue convergence rate is discussed.

  15. Establishment and preliminary application of an ELISA method for determination of tetanus toxoid%检测破伤风类毒素的酶联免疫吸附法的建立及应用

    Institute of Scientific and Technical Information of China (English)

    王玲; 王珣; 韩俊杰; 张建玲; 杨云凯; 张娜; 佟芳; 潘殊男; 张霖阳

    2016-01-01

    Objective To establish an ELISA method for quantitative determination of tetanus toxoid (TT ) for DTaP .Methods Rabbits were immunized with purified TT to prepare high‐titer serum antibodies against TT .Polyclonal anti‐TT antibodies were purified by octanoic acid‐ammonium sulfate precipitation method and labeled with horseradish peroxidase to establish a double antibody sandwich ELISA method .Results The ELISA method had good specificity for TT and no cross reactivities with filamentous hemagglutinin ,pertussis toxin or diphtheria toxoid .The best linearity of the ELISA method was within the range of 0 .5‐16 .0 Lf /L TT (R 2 > 0 .99) .The coefficient of variation and recover rates of intra‐ and inter‐assay were 4 .7% ‐9 .8% and 92 .7% ‐109 .0% ,respectively ,when 14 .0 ,12 .0 ,6 .0 ,3 .0 and 1 .5 Lf /L TT standards were detected ,and the precision and accuracy both met requirements of quality control . The detection limit of the ELISA method was 1 .5 Lf /L TT . Conclusion The established ELISA method can be applied to detecting TT during purification of tetanus vaccine and lays foundation for quality control of TT .%目的:建立定量检测白喉‐破伤风‐无细胞百日咳疫苗生产过程中破伤风类毒素(tetanus toxoid ,TT )的方法。方法 TT 免疫家兔以制备高效价血清抗 TT 抗体。辛酸‐硫酸铵沉淀法纯化抗TT 多克隆抗体并进行辣根过氧化物酶标记,建立双抗体夹心 ELISA 法。结果建立的 ELISA 法与丝状血凝素、百日咳毒素及白喉类毒素无交叉反应,特异性较好。该 ELISA 法在0.5~16.0 Lf /L TT 检测区间有最佳线性,决定系数>0.99。实验内和实验间检测14.0、12.0、6.0、3.0和1.5 Lf/L TT ,变异系数为4.7%~9.8%,回收率为92.7%~109.0%,精密度和准确度均符合常规质控要求。该法的定量下限为1.5 Lf/L TT 。结论建立的 ELISA 法可有效检测破

  16. Measuring Hordein (Gluten) in Beer – A Comparison of ELISA and Mass Spectrometry

    Science.gov (United States)

    Blundell, Malcolm J.; Goswami, Hareshwar P.; Howitt, Crispin A.

    2013-01-01

    Background Subjects suffering from coeliac disease, gluten allergy/intolerance must adopt a lifelong avoidance of gluten. Beer contains trace levels of hordeins (gluten) which are too high to be safely consumed by most coeliacs. Accurate measurement of trace hordeins by ELISA is problematic. Methods We have compared hordein levels in sixty beers, by sandwich ELISA, with the level determined using multiple reaction monitoring mass spectrometry (MRM-MS). Results Hordein levels measured by ELISA varied by four orders of magnitude, from zero (for known gluten-free beers) to 47,000 µg/mL (ppm; for a wheat-based beer). Half the commercial gluten-free beers were free of hordein by MS and ELISA. Two gluten-free and two low-gluten beers had zero ELISA readings, but contained significant hordein levels (pgluten in beverages such as beer and highlights the need for the development of new sensitive and selective quantitative assay such as MS. PMID:23509606

  17. Detection of Lupine (Lupinus spp. L. as a food allergen using three methods: end-point PCR, Real-Time PCR and Elisa

    Directory of Open Access Journals (Sweden)

    Ondrej Revák

    2014-07-01

    Full Text Available The aim of this work was to compare three methods for the detection and quantification of lupine as an allergen in food. The methods that were used in this work were the direct method: ELISA and the indirect methods: end-point PCR and real-time PCR. We examined the detection limit (the sensitivity with which we can detect the presence of the allergen in a sample and the reliability for performing an analysis. We used 17 samples of plant species from a processing plant for dehydrated soups production and lupine samples from lupine processing companies. Its practical use is wide and it is used mainly in the bakery industry, in the manufacture of confectionery, pasta, sauces, as a substitute for soy and also in the production of gluten-free food, because it does not contain gluten. Lupine, however, is also included in the list of 14 allergenic substances, which in accordance with the EU legislation must be listed on food labels. The high risk group, which suffers from primary sensitization or cross-reaction with peanuts, are allergic patients. In the EU, people who are allergic to peanuts range from 0.7 to 1.5%. In experiment 1, we detected the presence of lupine using primers for the detection of α- and δ-conglutine in the samples, using the end-point PCR method and the detection limit of this reaction was at the level of 100 ppm. For the vizualization of the DNA fragments, we used a 2% agarose gel and UV visualizer. In experiment 2 we detected lupine using the TaqMan real-time PCR reaction and primers for the detection of α and δ-conglutine at the level of 10 ppm of lupine in sample. The CP values of lupine using primers for the detection of α-conglutine was 24.85 ± 0.12 and the reliability equation was R2 = 0.9767. The CP lupine values using primers for the detection of δ-conglutine was 22.52 ± 0.17 and the reliability equation was R2 = 0.9925. In experiment 3, we used a sandwich ELISA method for the detection of lupine and the

  18. Entamoeba spp. diagnosis in patients with inflmmatory diarrhea by staining, copro-antigen ELISA and multiplex PCR methods

    Directory of Open Access Journals (Sweden)

    Zahra Gharibi

    2017-10-01

    Full Text Available Objective: To evaluate Entamoeba spp. diagnosis in patients with inflammatory diarrhea by staining, copro-antigen ELISA and multiplex PCR methods. Methods: In this descriptive cross-sectional survey, 200 stool samples were randomly collected during 2015–2016. The stool samples were evaluated microscopically for the presence of the parasite using direct and formalin-ether concentration and trichrome staining methods. Then, the stool samples were examined by copro-antigen ELISA (Biomerica Company and multiplex PCR methods. Results: Of 200 samples, 17, 29 and 23 cases were positive for Entamoeba species by the staining, copro-antigen ELISA and multiplex PCR methods, respectively. Of 23 positive samples in multiplex PCR test, 13 and 10 samples were positive for Entamoeba dispar (E. dispar and Entamoeba histolytica (E. histolytica, respectively. Conclusions: Our finding indicated a relatively high prevalence of Entamoeba species in patients with inflammatory diarrhea in Ahvaz city. Due to the complications of E. histolytica/ dispar infection, the health authorities of the city must pay more attention to control and prevent the transmission of E. histolytica/dispar to individuals.

  19. Predicting method of local damage in reinforced concrete plate with absorber sandwiched between two concrete panels under hard projectile impact

    Energy Technology Data Exchange (ETDEWEB)

    Shirai, Tetsuo; Kambayashi, Atsushi; Ueda, Masatoshi [Takenaka Corp., Osaka (Japan); Ohno, Tomonori; Ishikawa, Nobutaka

    1995-07-01

    The predicting method of local damage in reinforced concrete plate with absorber sandwiched between two concrete panels (double-layered RC plate) under the impact of hard projectile is studied in this paper. The results of high-velocity impact tests to investigate the impact resistance of double-layered RC plates are reported. To evaluate quantitatively the extent of local damage, the existing formulae and the alternative predicting formulae with the multivariate analysis presented here are employed. The prediction of the proposed formulae can agree reasonably well with the actual observed damage, thus can be a useful method in impact resistant design. (author).

  20. 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

  1. 2004 National Atrazine Occurrence Monitoring Program using the Abraxis ELISA method.

    Science.gov (United States)

    Graziano, Nicole; McGuire, Michael J; Roberson, Alan; Adams, Craig; Jiang, Hua; Blute, Nicole

    2006-02-15

    The goal of this project was to gain a better understanding of atrazine occurrence in the United States by surveying drinking water utilities' sources and finished water for atrazine on a weekly basis for seven months. Atrazine is a contaminant of interest because the United States Environmental Protection Agency (USEPA) has found short-term atrazine exposure above the drinking water maximum contaminant level (MCL) to potentially cause heart, lung, and kidney congestion, low blood pressure, muscle spasms, weight loss, and damage to the adrenal glands. Long-term exposure to atrazine concentrations above the drinking water MCL has been linked to weight loss, cardiovascular damage, retinal and muscle degeneration, and cancer. This survey effort improved upon previously conducted atrazine surveys through intensive, high frequency sampling (participating plants sampled their raw and finished water on a weekly basis for approximately seven months). Such an intensive effort allowed the authors to gain a better understanding of short-term atrazine occurrence and its variability in drinking water sources. This information can benefit the drinking water industry by facilitating (1) better atrazine occurrence management (i.e., awareness when plants may be more susceptible to atrazine), (2) more efficient atrazine control (e.g., effective treatment alternatives and more effective response to atrazine occurrence), and (3) treatment cost reduction (e.g., efficient atrazine control can result in substantial cost savings). Forty-seven drinking watertreatment plants located primarily in the Midwestern United States participated in the survey and sampled their raw and finished water on a weekly basis from March through October. Samples were analyzed using the Abraxis enzyme-linked immunosorbent assay (ELISA) test kit. Confirmation samples for quality assurance/quality control (QA/QC) purposes were analyzed using solid-phase extraction (SPE) followed by gas chromatography mass

  2. Research on a Novel Exciting Method for a Sandwich Transducer Operating in Longitudinal-Bending Hybrid Modes.

    Science.gov (United States)

    Liu, Yingxiang; Shen, Qiangqiang; Shi, Shengjun; Deng, Jie; Chen, Weishan; Wang, Liang

    2017-06-27

    A novel exciting method for a sandwich type piezoelectric transducer operating in longitudinal-bending hybrid vibration modes is proposed and discussed, in which the piezoelectric elements for the excitations of the longitudinal and bending vibrations share the same axial location, but correspond to different partitions. Whole-piece type piezoelectric plates with three separated partitions are used, in which the center partitions generate the first longitudinal vibration, while the upper and lower partitions produce the second bending vibration. Detailed comparisons between the proposed exciting method and the traditional one were accomplished by finite element method (FEM) calculations, which were further verified by experiments. Compared with the traditional exciting method using independent longitudinal ceramics and bending ceramics, the proposed method achieves higher electromechanical coupling factors and larger vibration amplitudes, especially for the bending vibration mode. This novel exciting method for longitudinal-bending hybrid vibrations has not changed the structural dimensions of the sandwich transducer, but markedly improves the mechanical output ability, which makes it very helpful and meaningful in designing new piezoelectric actuators operated in longitudinal-bending hybrid vibration modes.

  3. Frequency of antiphospholipid antibodies in patients with infectious diseases using three different ELISA methods Freqüência de anticorpos antifosfolípides em pacientes com doenças infecciosas usando três diferentes testes de ELISA

    Directory of Open Access Journals (Sweden)

    Mittermayer Barreto Santiago

    2006-02-01

    Full Text Available OBJECTIVE: The standard enzyme-linked immunosorbent assay (ELISA for anticardiolipin (aCL antibodies is the most important test for the diagnosis of antiphospholipid syndrome (APS. However, the test is also positive in some infectious diseases and other non-related syndromes. It has been suggested that the detection of antibodies to a mixture of phospholipids or to beta2-glycoprotein I (beta2-GP I has higher specificity for APS than the standard aCL ELISA. The aim of the present work is to compare the diagnostic specificity of three different antiphospholipid (aPL assays in patients with infectious diseases. METHODS: Antiphospholipid antibodies were searched by three ELISA techniques, namely standard aCL, APhL® ELISA kit and anti-beta2-GP I, in sera of patients with infectious diseases, including syphilis (69, leptospirosis (33 and visceral leishmaniasis (30. RESULTS: The frequency of positivity of IgG aPL in patients with syphilis, leptospirosis and Kala-azar was 13/69 (19%, 9/33 (27% and 2/30 (6%, respectively, using standard ELISA, versus only 1/69 (1.4%, 0/33 (0% and 0/30 (0% positivity by the APhL® ELISA kit. The positivity of the isotype IgM aPL was 10/69 (14%, 4/33 (12% and 1/30 (3%, respectively, by the standard ELISA, and 1/69 (1.4%, 0/33 (0% and 0/30 (0% by the APhL® ELISA kit. The presence of significant levels of IgG anti-beta2GPI was observed in 14/69 cases of syphilis (20%, 6/33 cases of leptospirosis (18% and 16/30 cases of Kala-azar (53%. The APhL® ELISA kit had superior performance showing the highest specificity: 97% (95% CI: 92%-99% for IgG compared to 81% (95% CI: 74%-87% for standard ELISA and 72% (95% CI: 64%-79% for anti-beta2 GPI assay. CONCLUSIONS: The APhL® ELISA kit proved to be significantly more specific than the aCL standard ELISA and the anti-beta2GPI ELISA, and it should be used to help in the diagnosis and confirmation of APS.OBJETIVO: O ensaio de enzyme-linked immunosorbent assay (ELISA para a pesquisa de

  4. Structural and failure mechanics of sandwich composites

    CERN Document Server

    Carlsson, LA; Carlsson, Leif A

    2011-01-01

    Focusing on important deformation and failure modes of sandwich structures, this volume describes the mechanics behind fracture processes. The text also reviews test methods developed for the cr, structural integrity, and failure mechanisms of sandwich structures.

  5. Establishment of a sandwich ELISA for quantitative measurement of human supersensitivity C-reactive protein and primary clinical applica-tion study%定量检测人超敏C-反应蛋白双抗体夹心ELISA方法的建立及初步临床应用

    Institute of Scientific and Technical Information of China (English)

    沈丹丹; 卞智萍; 何国平; 顾春荣; 徐晋姚; 杨笛; 张寄南

    2009-01-01

    AIM: To establish a sandwich ELISA for quantitative measurement of supersensitivity C-reac-tive protein(hs-CRP). METHODS: Anti-CRP monoclonal antibodies (mAbs) prepared by our laboratory were purified by Protein A affinity chromatography and analyzed by SDS-PAGE and Western-blotting to test their characteristics. All the mAbs were labeled with horseradish peroxidase by sodium oxidation method and antibody mating test were performed using anti-CRP mAb as coating antibody and HRP labeled anti-CRP mAb as labeled antibody, in which optimal concentrations were defined by square mateix titration. Standard curve was performed using purified CRP and the sensitivity, reproducibility and recovery rate test of ELISA was evaluated. The CRP levels in plasma were measured in this assay. RESULTS: The optimal paired antibodies were anti-CRP mAb 1C10 and HRP labeled anti-CRP mAb 2C11, and the optimal concentrations were 10 μg/mL and 1: 2000,respectively. The coefficient of variation were 3.1% to 9.7 % within assay and 3.6% to 13.6% between assay. The sensitivity in this assay was 8.3 ng/mL. The recovery rate was 90% to 109%. According to the result,68 normal persons' hs-CRP level in plasma of coronary arteriography were detected by ELISA, 59 with stenosis < 50% and 67 with stenosis ≥50%. The results showed that the plasma hs-CRP level in patients with stenosis < 50% (3.65 ± 1.15) mg/L was significantly higher ( P < 0.05)than hs-CRP in normol persons(1.75 ± 0.74) mg/L, the plasma hs-CRP level in stenosis ≥ 50% patients ( 8.93 ± 3.29) mg/L was significantly higher (P < 0.05) than stenosis < 50% patients. CONCLUSION: A sandwich ELISA for detecting hs-CRP was established.%目的:建立定量检测人超敏C-反应蛋白(CRP)双抗体夹心ELISA方法.方法:采用ProteinA亲和层析法纯化本室制备的抗CRP单克隆抗体(mAbs),并行SDS-PAGE和Western blotting对纯化抗体特性进行鉴定;利用简易过碘酸钠法对抗CRP mAbs进行标记HRP后行抗体配对实

  6. Accuracy of ELISA detection methods for gluten and reference materials: a realistic assessment.

    Science.gov (United States)

    Diaz-Amigo, Carmen; Popping, Bert

    2013-06-19

    The determination of prolamins by ELISA and subsequent conversion of the resulting concentration to gluten content in food appears to be a comparatively simple and straightforward process with which many laboratories have years-long experience. At the end of the process, a value of gluten, expressed in mg/kg or ppm, is obtained. This value often is the basis for the decision if a product can be labeled gluten-free or not. On the basis of currently available scientific information, the accuracy of the obtained values with commonly used commercial ELISA kits has to be questioned. Although recently several multilaboratory studies have been conducted in an attempt to emphasize and ensure the accuracy of the results, data suggest that it was the precision of these assays, not the accuracy, that was confirmed because some of the underlying assumptions for calculating the gluten content lack scientific data support as well as appropriate reference materials for comparison. This paper discusses the issues of gluten determination and quantification with respect to antibody specificity, extraction procedures, reference materials, and their commutability.

  7. Analysis of questionnaire of Sandwich teaching method%Sandwich教学法的应用调查问卷分析

    Institute of Scientific and Technical Information of China (English)

    何姗姗; 谢莎; 石焕焕; 李艳文; 刘登宇; 唐莉莉; 战廷正

    2015-01-01

    In order to inspire the subjective initiative of students and increase the comprehensive quality of teachers as well as students, the author of this thesis used the "Sandwich" method in the teaching of Medical Parasitology and collected students' opinions in the form of questionnaires for teaching evaluation, which indicates that the "Sandwich" method is a practical method that could accommodate to the current situation of basic medical education.%为了发挥学生的主观能动性,充分体现其主体作用,提高教师与学生综合素质。笔者在《医学寄生虫学》教学中应用Sandwich教学法,并收集学生调查问卷进行教学评价。分析结果显示:Sandwich教学法是一种较适应当前基础医学教育实情的教学方法。

  8. A novel colorimetric sandwich aptasensor based on an indirect competitive enzyme-free method for ultrasensitive detection of chloramphenicol.

    Science.gov (United States)

    Abnous, Khalil; Danesh, Noor Mohammad; Ramezani, Mohammad; Emrani, Ahmad Sarreshtehdar; Taghdisi, Seyed Mohammad

    2016-04-15

    Analytical methods for detection and quantitation of chloramphenicol in blood serum and foodstuffs arse highly in demand. In this study, a colorimetric sandwich aptamer-based sensor (aptasensor) was fabricated for sensitive and selective detection of chloramphenicol, based on an indirect competitive enzyme-free assay using gold nanoparticles (AuNPs), biotin and streptavidin. The designed aptasensor acquires characteristics of AuNPs, including large surface area and unique optical properties, and strong interaction of biotin with streptavidin. In the absence of chloramphenicol, the sandwich structure of aptasensor forms, leading to the observation of sharp red color. In the presence of target, functionalized AuNPs could not bind to 96-well plates, resulting in a faint red color. The fabricated colorimetric aptasensor exhibited high selectivity toward chloramphenicol with a limit of detection as low as 451 pM. Moreover, the developed colorimetric aptasensor was successfully used to detect chloramphenicol in milk and serum with LODs of 697 and 601 pM, respectively.

  9. Development of ELISA-based methods to measure the anti-malarial drug chloroquine in plasma and in pharmaceutical formulations

    Directory of Open Access Journals (Sweden)

    Ronn Anita

    2011-08-01

    Full Text Available Abstract Background In Central and South America and Eastern and Southern Africa, Plasmodium vivax infections accounts for 71-81% and 5% of malaria cases, respectively. In these areas, chloroquine (CQ remains the treatment of choice for P. vivax malaria. In addition, CQ has recently proven to be an effective HIV-1 therapeutic agent. There is a dire need to continue monitoring quality of CQ as there is a major influx of substandard and fake formulations into malaria-endemic countries. The use of fake/substandard drugs will result in sub-therapeutic levels endangering the patient and possibly select for parasite resistance. The aim of this study was to develop an inexpensive, simple antibody-based ELISA to measure CQ concentrations in tablets and in plasma. Methods A monoclonal antibody (MAb that reacts with the N-side chain of the CQ molecule was prepared by use of a CQ analogue. A specific and reliable ELISA for detection of CQ was developed. The developed assay was validated by measuring CQ in tablets sold in Denmark, India and Sudan. Furthermore, kinetics of CQ concentrations in plasma of four volunteers, who ingested two tablets of Malarex® containing, 250 mg CQ base, were measured before drug intake, three hours later and thereafter at days 1, 3, 7, 14, 21 and 28. The same plasma samples were simultaneously measured by high performance liquid chromatography (HPLC. Results The ELISA proved an easy-to-handle and very sensitive tool for the detection of CQ with a lower limit of detection at 3.9 ng/ml. ELISA levels of CQ in plasma showed high agreement with the levels obtained by HPLC (r = 0.98. The specificity in the negative control group was 100%. Conclusion The developed ELISA can be used for quality screening of CQ in pharmaceutical formulations and for drug monitoring in malaria and in other infectious diseases, such as HIV, where CQ proved to be an effective therapeutic agent. The methodology has been exploited to develop monoclonal

  10. The detection of specific immune complex by using double McAb sandwich ELISA in sera of patients with neurocysticercosis%双单抗夹心ELISA法检测脑囊尾蚴病血清特异性免疫复合物

    Institute of Scientific and Technical Information of China (English)

    张明霞; 林尔昕; 王淑民; 崔琢

    2001-01-01

    Objective:To develop a double monoclonal an tibody sandwich ELISA assay for detecting specific immune complex(IC) in sera of patients with neurocysticercosis.Methods:Anti-human IgG McAb was employed as coat,and HRP labeled an ti-cysticerci McAb as recognizing system to detect antibody specific IC.Results:The IC positive rate was found in 53.3% of patients(P <0.005).By using the new method to detect three different kinds of serological markers,the results showed the IC positive rates for group of CAg(-) and CAb(-) was 50%(P<0.005).The IC detection rate was higher than CAg's(P<0. 005).Conclusions:It suggests that detection of IC is of significance in i mproving diagnosis in neurocysticercosis,especially with CAg(-) and CAb(-).%目的:建立双单抗夹心酶联免疫吸附试验法检测脑囊尾蚴病患者血清中特异性免疫复合物。方法:用抗人IgG单抗包被,捕获血清中特异性IgG型免疫复合物,通过酶标抗囊尾蚴单抗结合物显色,测其OD值。结果:用本方法测得患者特异性免疫复合物阳性率为53.3%,明显高于正常对照组(P<0.005);循环抗原、抗体皆阴性患者,免疫复合物的阳性率为50%,与各组相比差异有显著性(P<0.005);且免疫复合物的检出率明显高于循环抗原的检出率( P<0.005)。结论:本法对囊尾蚴病的诊断有重要的临床价值,尤其对抗原、抗体皆阴性患者意义更大。

  11. Direct ELISA kits as a sensitive and selective screening method for abstinence control in urine.

    Science.gov (United States)

    Kirschbaum, Katrin M; Musshoff, Frank; Wilbert, Ansgar; Röhrich, Jörg; Madea, Burkhard

    2011-04-15

    In 2009 cutoff values of assessment criteria to testify abstinence control in order to estimate driving ability were standardized in Germany. The cutoff values are lower than required in existing guidelines like SAMHSA and there is critical discussion about detection of low concentrations by using immunoassay, especially concerning amphetamines in urine (50 ng/ml). In this study Direct ELISA kits were tested for their applicability to identify the absence of amphetamines, cannabinoids, opiates, cocaine, methadone and benzodiazepines in urine. Results were confirmed by LC/MS or GC/MS analyses. Sensitivity, specificity, predictive values (positive as well as negative) and overall misclassification rates were evaluated by contingency tables and were compared to ROC-analyses. Sensitivity results as well as specificity results were satisfying showing sensitivity values higher than 96% for each analyte. The amphetamine test we used showed sensitivity and specificity of 100% and 88%, respectively, even if amphetamine tests usually react with high cross-reactivity. Our study results include high discrimination at required cutoff values between positives and negatives for each drug group and demonstrate that immunological tests complying with requirements of current decreased urine cutoff values for assessment of driving ability do exist.

  12. Setting up of a liquid chromatography-high resolution tandem mass spectrometry method for the detection of caseins in food. A comparison with ELISA method

    Directory of Open Access Journals (Sweden)

    Daniela Gastaldi

    2013-06-01

    Full Text Available Determination of caseins in food matrices is usually performed by using the competitiveenzyme- linked immunosorbent assay (ELISA technique. However such a technique suffers from a number of limitations. Among these, the applicability to a narrow concentration range, a non linear (logarithmic response, a non-negligible cross-reactivity and a high cost per kit. At the time of the completion of this study, in case of ELISA positive feedback, there was poor availability in the literature of finding reliable instrumental methods able to determine both qualitatively and quantitatively this class of substances. In the present study, a liquid chromatography-high resolution tandem mass spectrometry (HPLC-HRMS/MS instrumental method was developed with a high resolution mass spectrometer (Orbitrap. Real samples of sausages in which caseins were detected by ELISA technique were analysed. A casein-free sample of ham was used as a blank. The analytical characteristics of the instrumental method were compared with those of a commercial ELISA test, declared specific for α- and β-casein.

  13. [Surgical Treatment of Large Muscular Ventricular Septal Defect nearby the Moderator Band Using the Sandwich Method;Report of a Case].

    Science.gov (United States)

    Motokawa, Mamika; Sasahara, Akihiro; Terakawa, Katsunari; Miyamoto, Takashi

    2016-09-01

    We describe the rare case of a 1-year-old girl who had large muscular ventricular defect (VSD) nearby the moderator band. We experienced the patch closure using sandwich method. A 1-month-old girl was referred to our institution for treatment of muscular VSD. At the age of 2 month, she underwent the pulmonary artery banding to control the pulmonary high flow. After follow up, the patient have reached 70 cm tall and weighed 7 kg. One year after the "sandwich operation", cardiac catheterization revealed the tiny residual shunt. Nevertheless, the cardiac function was good and the growth was in fine fettle. Sandwich method is a useful surgical technique to close the muscular VSD without resect the right ventricular trabeculation.

  14. Identification of tylosin photoreaction products and comparison of ELISA and HPLC methods for their detection in water.

    Science.gov (United States)

    Hu, Dingfei; Fulton, Bruce; Henderson, Keri; Coats, Joel

    2008-04-15

    Tylosin is a widely used macrolide antibiotic for therapeutics and growth promotion in swine, beef cattle, and poultry production. Through various routes such as manure application, emission, inappropriate disposal, etc., tylosin enters the environment. The fate of tylosin in the environment is not yet fully understood. In this study, two photoreaction products of tylosin in water were identified as isotylosin A alcohol (E,Z) and isotylosin A aldol (E,Z). Tylosin A, B, C, D, isotylosin A alcohol, and isotylosin A aldol were purified, and immunological cross-reactivities of these tylosin-related compounds were tested with a specificity of 26% for tylosin B, 19% for tylosin C, 106% for tylosin D, 121% for isotylosin A alcohol, and 46% for isotylosin A aldol, compared to 100% for tylosin A. Competitive direct enzyme-linked immunosorbent assay (ELISA) for tylosin detection in water was compared with a high-performance liquid chromatography (HPLC) method by analyzing the same water samples from a study of tylosin dissipation in water. ELISA kits detect the other tylosin-related compounds besides tylosin A, which can result in differences in tylosin determination in water.

  15. Characterization of a Toxocara canis species-specific excretory-secretory antigen (TcES-57) and development of a double sandwich ELISA for diagnosis of visceral larva migrans

    OpenAIRE

    Iddawela, R.D.; R.P.V.J Rajapakse; Perera, N.A.N.D.; Agatsuma, Takeshi

    2007-01-01

    This study describes the isolation of a Toxocara canis species-specific excretory-secretory (ES) antigen and the development of an enzyme-linked immunosorbent assay (ELISA) based on this antigen. Analysis of the ES antigens of T. canis, Toxocara vitulorum, Ascaris lumbricoides and Necator americanus larval antigen was performed by SDS-PAGE followed by western blotting. A 57 kDa T. canis-specific antibody fraction (TcES-57) was identified by western blotting and labelling with anti-Toxocara an...

  16. Evaluation of modified ELISA method to detect Hepatitis B surface antigen in saliva specimens%ELISA 检测唾液中乙肝表面抗原方法的优化与评价

    Institute of Scientific and Technical Information of China (English)

    张进

    2012-01-01

      目的优化用于检测唾液中乙肝表面抗原的 ELISA 法,并对其进行评价.方法对 ELISA 的不同条件:样品类型、加样体积和孵育温度及时间进行优化,筛选最佳条件;并采用三种不同的临界值计算方法,判断最佳临界值.结果采用优化后方法,47例血清阳性对应的唾液样品中,44例检测结果为阳性;68例血清阴性对应的唾液样品中,63例检测结果为阴性.血清和唾液样品检测结果的一致性比较高,κ指数为0.87.且该方法的特异度和敏感度分别达到92.6%和93.6%.结论优化后的 ELISA 方法在唾液样品的乙肝表面抗原检测中具有潜在的应用价值.%  Objective To evaluated a modified ELISA method for detecting the Hepatitis B surface antigen (HBsAg) in saliva samples. Meyhods Optimized the different conditions, including sample type, sample volume and incubation condition, to screen the best condition. Three different methods to calculate cut-off value was evaluated to screen the most acceptable. Results HBsAg was detected in 44 saliva samples out of 47 paired positive serum specimens and not detected in 63 saliva samples out of 68 matched negative serum samples by the optimized ELISA assay. There was excelent agreement between the results for the serum and saliva specimens and the kappa value was 0.87 for saliva specimens. Using an optimized protocol, the sensitivities and specificities were 93.6% and 92.6%, respectively. Conclusion Our data showed a significant promise for the use of the modified commercial ELISA in saliva sample for Hepatitis B virus infection surveilance.

  17. Food allergen analysis for processed food using a novel extraction method to eliminate harmful reagents for both ELISA and lateral-flow tests.

    Science.gov (United States)

    Ito, Kaori; Yamamoto, Takayuki; Oyama, Yuriko; Tsuruma, Rieko; Saito, Eriko; Saito, Yoshikazu; Ozu, Takeshi; Honjoh, Tsutomu; Adachi, Reiko; Sakai, Shinobu; Akiyama, Hiroshi; Shoji, Masahiro

    2016-09-01

    Enzyme-linked immunosorbent assay (ELISA) is commonly used to determine food allergens in food products. However, a significant number of ELISAs give an erroneous result, especially when applied to highly processed food. Accordingly, an improved ELISA, which utilizes an extraction solution comprising the surfactant sodium lauryl sulfate (SDS) and reductant 2-mercaptoethanol (2-ME), has been specially developed to analyze food allergens in highly processed food by enhancing analyte protein extraction. Recently, however, the use of 2-ME has become undesirable. In the present study, a new extraction solution containing a human- and eco-friendly reductant, which is convenient to use at the food manufacturing site, has been established. Among three chemicals with different reducing properties, sodium sulfite, tris(3-hydroxypropyl)phosphine, and mercaptoethylamine sodium sulfite was selected as a 2-ME substitute. The protein extraction ability of SDS/0.1 M sodium sulfite solution was comparable to that of SDS/2-ME solution. Next, the ELISA performance for egg, milk, wheat, peanut, and buckwheat was evaluated by using model-processed foods and commercially available food products. The data showed that the SDS/0.1 M sulfite ELISA significantly correlated with the SDS/2-ME ELISA for all food allergens examined (p food allergens in processed food, showing consistency with the SDS/0.1 M sulfite ELISA results. Accordingly, a harmonized analysis system for processed food comprising a screening LF test and a quantitative ELISA with identical extraction solution has been established. The ELISA based on the SDS/0.1 M sulfite extraction solution has now been authorized as the revised official method for food allergen analysis in Japan.

  18. Preparation of double-antibody and establishment of sandwich ELISA against milk β-casein and soybeanβ-conglycinin . Journal of Zhejiang University (Agric . & Life Sci .), 2013,39(2):222-226%牛奶β-酪蛋白和大豆β-伴球蛋白双抗制备及夹心ELISA快速定性检测技术的建立

    Institute of Scientific and Technical Information of China (English)

    肖海龙; 赵凯; 林赛君; 王红青; 潘建红

    2013-01-01

    Summary Protein adulteration and allergens are the two major food safety issues , and can pose health threats to consumers . One of the effective precautions is extensive test , which needs simple , rapid and low‐cost test method . Present methods including sodium dodecyl sulfate polyacrylamide gel electrophoresis ( SDS‐PAGE) , high performance liquid chromatography ( HPLC) and liquid chromatography‐mass spectrometry ( LC‐MS)/MS are not acceptable for consumers due to expensive instruments . Immunological technique is a rapid method for screening and is suitable for consumers to use .β‐casein andβ‐conglycinin are not only the major proteins of milk and soybean , but also important food borne allergens . In this study , the monoclonal antibodies and polyclonal antibodies against β‐casein and β‐conglycinin were prepared and the enzyme‐linked immunosorbent assay ( ELISA ) kits were established for detecting adulteration and allergens . BALB/c mice were immunized four times with purified antigens adding adjuvant or not until the serum titer achieved 1∶1 × 105 , and the mice spleen cells and myeloma cells SP2/0 were fused as the routine cell‐fusion technology . Positive cells were screened for 3‐4 times with indirect ELISA by coating purified antigens , and were injected into mice peritoneal . The monoclonal antibodies were obtained after the purification of ascites . The polyclonal antibodies against β‐casein and β‐conglycinin were also prepared from rabbit serum immunized by antigens , respectively . The double antibody sandwich ELISA for β‐casein and β‐conglycinin were successfully established by optimized parameters . The results showed that the titers of purified monoclonal antibodies of β‐casein and β‐conglycinin were over 1∶1 × 107 , and the polyclonal antibody titers of both were about 1∶2 × 105 . The minimum detection limits of both ELISA kits were about 15 ng/mL , and no cross reaction were observed among

  19. Rapid screen for truncating ATM mutations by PTT-ELISA

    Energy Technology Data Exchange (ETDEWEB)

    Du Liutao; Lai, C.-H. [Department of Pathology and Laboratory Medicine, David Geffen School of Medicine at UCLA, CA 90095 (United States); Concannon, Patrick [Department of Biochemistry and Molecular Genetics, University of Virginia, VA 22908 (United States); Gatti, Richard A. [Department of Pathology and Laboratory Medicine, David Geffen School of Medicine at UCLA, CA 90095 (United States); Department of Human Genetics, David Geffen School of Medicine at UCLA, CA 90095 (United States)], E-mail: rgatti@mednet.ucla.edu

    2008-04-02

    Mutations in the ataxia-telangiectasia mutated (ATM) gene are responsible for the autosomal recessive genetic disorder, ataxia-telangiectasia (A-T). Approximately 80% of ATM mutations found in A-T patients results in truncations, which can be detected by Protein Truncation Test (PTT). Conventional PTT uses SDS-PAGE electrophoresis to detect mobility of radiolabeled truncated protein fragments. In this study, we developed a non-radioactive Protein Truncation Test which utilizes an enzyme-linked immunosorbent assay (PTT-ELISA) to detect ATM mutations in eight overlapping fragments. N- and C-terminal epitopes (c-myc and V5, respectively) were introduced into transcription/translation products, which could then be detected by Sandwich ELISA. Using this assay, we screened 9 newly diagnosed A-T patients consecutively. Of the 18 expected mutations, 14 truncating mutations were independently identified by cDNA direct sequencing and/or DNA dHPLC analysis. PTT-ELISA detected all of these 14. Four mutations were novel. The PTT-ELISA provides a rapid method for detecting truncating mutations in large genes and should be considered prior to using more laborious or costly methods, such as direct sequencing.

  20. Measuring hordein (gluten in beer--a comparison of ELISA and mass spectrometry.

    Directory of Open Access Journals (Sweden)

    Gregory J Tanner

    Full Text Available BACKGROUND: Subjects suffering from coeliac disease, gluten allergy/intolerance must adopt a lifelong avoidance of gluten. Beer contains trace levels of hordeins (gluten which are too high to be safely consumed by most coeliacs. Accurate measurement of trace hordeins by ELISA is problematic. METHODS: We have compared hordein levels in sixty beers, by sandwich ELISA, with the level determined using multiple reaction monitoring mass spectrometry (MRM-MS. RESULTS: Hordein levels measured by ELISA varied by four orders of magnitude, from zero (for known gluten-free beers to 47,000 µg/mL (ppm; for a wheat-based beer. Half the commercial gluten-free beers were free of hordein by MS and ELISA. Two gluten-free and two low-gluten beers had zero ELISA readings, but contained significant hordein levels (p<0.05, or near average (60-140% hordein levels, by MS, respectively. Six beers gave false negatives, with zero ELISA readings but near average hordein content by MS. Approximately 20% of commercial beers had ELISA readings less than 1 ppm, but a near average hordein content by MS. Several barley beers also contained undeclared wheat proteins. CONCLUSIONS: ELISA results did not correlate with the relative content of hordein peptides determined by MS, with all barley based beers containing hordein. We suggest that mass spectrometry is more reliable than ELISA, as ELISA enumerates only the concentration of particular amino-acid epitopes; this may vary between different hordeins and may not be related to the absolute hordein concentration. MS quantification is undertaken using peptides that are specific and unique, enabling the quantification of individual hordein isoforms. This outlines the problem of relying solely on ELISA determination of gluten in beverages such as beer and highlights the need for the development of new sensitive and selective quantitative assay such as MS.

  1. Development of a Double-antibody Sandwich ELISA for Detection of Subgroup J Avian Leukosis Virus%J亚群禽白血病病毒双抗体夹心ELISA检测方法的建立

    Institute of Scientific and Technical Information of China (English)

    廖亚琳; 梁艺瑜; 王秀珑; 冯敏; 谭利强; 曹伟胜

    2013-01-01

    本研究利用J亚群禽白血病病毒(subgroup J avian leukosis virus,ALV-J)gp85单因子血清纯化后的抗体成功建立了检测ALV-J抗原的双抗体夹心ELISA方法(DAS-ELISA).结果表明,该方法具有良好的特异性、重复性和稳定性,对ALV-J抗原的最小检出量为0.165 μg/mL.用该法对48份临床血浆样品进行检测,结果与PCR方法的符合率达到85.2%.

  2. Determination of aflatoxins in nuts of Tabriz confectionaries by ELISA and HPLC methods

    OpenAIRE

    Siahi Shadbad Mohammad Reza; Ansarin Masoud; Tahavori Ali; Ghaderi Faranak; Nemati Mahboob

    2012-01-01

    Purpose: Aflatoxins (AFs) are a group of mycotoxins and secondary metabolites of various species of Aspergillus. There are various forms of aflatoxins including B1, B2, G1, G2, M1 and M2 types. Aflatoxins cause important health problems and have high potential effect on liver cancer. Therefore, numerous investigations have been conducted during last three decades. The aim of this work is to determine the contamination levels of nuts used by the confectionaries in Tabriz. Methods: A total of 1...

  3. 肾综合征出血热IMMS-ELISA检测方法建立%IMMS-ELISA :a test method for hemorrhagic fever with renal syndrome

    Institute of Scientific and Technical Information of China (English)

    苏旭; 赵凤玲; 吕莉琨; 李力; 杨东靖; 陈锦英

    2012-01-01

    目的 建立一种用于检测肾综合征出血热(HFRS)特异性抗体IgG的ELISA方法一免疫磁性微球(IMMS)ELISA.方法 利用重组菌株E.coli BL21( DE3) /pET32a-L99S诱导表达SEO型汉坦病毒(HV)重组核蛋白(rNP),并进行镍亲和层析纯化,以纯化rNP为抗原,分别建立检测HFRS特异性抗体IgG的3种ELISA法:间接ELISA、捕获ELISA和IMMS -ELISA,并进行方法学比较.结果 新建立的3种ELISA的检测灵敏度和特异度为≥90%,总符合率≥95%.其中间接ELISA的灵敏度达100%,而假阳性率为10%;捕获ELISA特异度达100%,灵敏度较低(92.5%),且假阴性率为7.5%;IMMS-ELISA的灵敏度和特异度均达到100%.结论 IMMS-ELISA较其他2种ELISA检测方法更为简单、安全和准确,易于在基层公共卫生和临床医疗机构推广使用.%Objective To establish a novel enzyme-linked immunosorbent assay(ELISA) method, namely immu-nomagnetic microspheres( IMMS) -ELISA, to detect specific IgG in hemorrhagic fever patients with renal syndrom( HFRS). Methods The recombinant nucleocapsid protein(rNP) of SEO hantavirus was expressed by the recombinant strain,E. Coli BL21(DE3)/pET32a-L99S. Purified rNP was used as an antigen to establish three ELISA methods, including indirect ELISA,capture ELISA and IMMS-ELISA,for the detection of specific IgG in HFRS patients. The results of the three methods were compared with each other. Results The sensitivity and specificity of the three ELISA methods were all higher than 90% and their coincidence rates were all higher than 95%. The sensitivity of indirect ELISA was 100% ,but its false positive rate was 10%. The specificity of capture ELISA was 100% ,but its sensitivity was lower(92. 5% ) than that of other two methods. The sensitivity and specificity of IMMS-ELISA were all 100%. Conclusion IMMS-ELISA is a simpler, safer,and more accurate than indirect and capture ELISA method, and easy to use in grassroots public health and clinical medical

  4. Quantifying and Reducing Uncertainty in Estimated Microcystin Concentrations from the ELISA Method.

    Science.gov (United States)

    Qian, Song S; Chaffin, Justin D; DuFour, Mark R; Sherman, Jessica J; Golnick, Phoenix C; Collier, Christopher D; Nummer, Stephanie A; Margida, Michaela G

    2015-12-15

    We discuss the uncertainty associated with a commonly used method for measuring the concentration of microcystin, a group of toxins associated with cyanobacterial blooms. Such uncertainty is rarely reported and accounted for in important drinking water management decisions. Using monitoring data from Ohio Environmental Protection Agency and from City of Toledo, we document the sources of measurement uncertainty and recommend a Bayesian hierarchical modeling approach for reducing the measurement uncertainty. Our analysis suggests that (1) much of the uncertainty is a result of the highly uncertain "standard curve" developed during each test and (2) the uncertainty can be reduced by pooling raw test data from multiple tests. Based on these results, we suggest that estimation uncertainty can be effectively reduced through the effort of either (1) regional regulatory agencies by sharing and combining raw test data from regularly scheduled microcystin monitoring program or (2) the manufacturer of the testing kit by conducting additional tests as part of an effort to improve the testing kit.

  5. Detection of syphilis antibody TRUSE method and ELISA method%TRUSE法与ELISA法检测梅毒抗体的比较

    Institute of Scientific and Technical Information of China (English)

    崔金兰

    2016-01-01

    目的:分析两种梅毒检测方法的临床意义。方法:应用酶联免疫吸附试验(ELISA)和梅毒甲苯胺红不加热血清试验(TRUSE)对北京市某医院2015年6月~9月间对新入所的1247例人员进行的抗TP抗体检测。比较分析ELISA法与TRUSE法检测的结果。结果:1247例标本中ELISA阳性病例86例,感染率6.89%;胶体金法阳性病例61例,感染率4.89%;TURSE法阳性病例27,感染率2.16%。以胶体金法结果为金标准,对TRUSE法与ELISA法检测梅毒抗体的结果进行临床诊断性试验的比较,TURSE法检测阳性率低于ELISA(P<0.05)。结论:TURSE法检测灵敏度比ELISA低,并且受人为因素影响较大,不适用于梅毒螺旋体感染初筛的检测。ELISA检测梅毒螺旋体抗体的敏感性和特异性较高,适用与临床大样本量筛查。%Objective: to analyze the clinical significance of the detection methods of two kinds of syphilis. Methods: using the enzyme-linked immune adsorption assay (ELISA) and Syphilis toluidine red unheated serum test (TRUSE) detection in a hospital in Beijing by 2015 from 6 months to 9 months to new into the 1247 cases of the anti TP antibody detection. Comparative analysis of the results of ELISA reagents and coloidal gold detection. Results: 1247 cases of ELISA positive cases in 86 cases, the infection rate was 6.89%; 61 cases of coloid gold positive cases, the infection rate was 4.89%; TRUSE positive cases 27, the infection rate was 2.16%. With coloidal gold method as a gold standard, the results of TRUSE method and ELISA method for the detection of syphilis antibody clinical diagnostic tests are compared.The positive rate of TRUSE was lower than that of ELISA ( P<0.05). . Conclusion: TRUSE assay sensitivity than ELISA is low, and by human factors influence is big, is not applicable in the early infection of Treponema palidum sieve detection. The sensitivity and specificity of ELISA for the detection of syphilis

  6. Optimal locations of piezoelectric patches for supersonic flutter control of honeycomb sandwich panels, using the NSGA-II method

    Science.gov (United States)

    Nezami, M.; Gholami, B.

    2016-03-01

    The active flutter control of supersonic sandwich panels with regular honeycomb interlayers under impact load excitation is studied using piezoelectric patches. A non-dominated sorting-based multi-objective evolutionary algorithm, called non-dominated sorting genetic algorithm II (NSGA-II) is suggested to find the optimal locations for different numbers of piezoelectric actuator/sensor pairs. Quasi-steady first order supersonic piston theory is employed to define aerodynamic loading and the p-method is applied to find the flutter bounds. Hamilton’s principle in conjunction with the generalized Fourier expansions and Galerkin method are used to develop the dynamical model of the structural systems in the state-space domain. The classical Runge-Kutta time integration algorithm is then used to calculate the open-loop aeroelastic response of the system. The maximum flutter velocity and minimum voltage applied to actuators are calculated according to the optimal locations of piezoelectric patches obtained using the NSGA-II and then the proportional feedback is used to actively suppress the closed loop system response. Finally the control effects, using the two different controllers, are compared.

  7. Biotin-avidin sandwich elisa with specific human isotypes IgG1 and IgG4 for Culicidae mosquito blood meal identification from an epizootic yellow fever area in Brazil

    Directory of Open Access Journals (Sweden)

    AM Marassá

    2009-01-01

    Full Text Available With a view toward investigating the feeding behavior of Culicidae mosquitoes from an area of epizootic yellow fever transmission in the municipalities of Garruchos and Santo Antônio das Missões, Rio Grande do Sul State, Brazil, specimens were collected by aspiration from September 2005 to April 2007. The engorged females were submitted to blood meal identification by enzyme-linked immunosorbent assay (ELISA. A total of 142 blood-engorged samples were examined for human or monkey blood through species-specific IgG. Additional tests for specificity utilizing isotypes IgG1 and IgG4 of human monoclonal antibodies showed that only anti-human IgG1 was effective in recognizing blood meals of human origin. The results indicated a significant difference (p = 0.027 in detection patterns in samples of Haemagogus leucocelaenus recorded from human blood meals at Santo Antônio das Missões, which suggests some degree of exposure, since it was an area where epizootic outbreaks have been reported.

  8. 双抗体夹心ELISA检测HIV-1 gp41抗原方法的建立%Establishment of Antibody-Sandwich ELISA for Detection of HIV-1 gp41 Antigen

    Institute of Scientific and Technical Information of China (English)

    于澜; 徐志凯; 王海涛; 黎志东; 张芳琳

    2006-01-01

    目的:建立检测HIV-1 gp41抗原的双抗体夹心ELISA,并探讨其临床应用的可行性.方法:用饱和硫酸铵(SAS)纯化抗HIV-1 gp41-5单克隆抗体(mAb),用HRP标记后建立双抗体夹心ELISA法,对其灵敏度及特异性进行检测,并用该方法对40份HIV-1阳性血清进行了检测.结果:用mAb E12(5μg/mL)为包被抗体,2H6为酶标记抗体(1:900)建立了双抗体夹心ELISA法,检测gp41-5多肽的灵敏度是100 pg/mL.对HIV-1阳性血清中gp41抗原的检出率为67.5%(27/40).结论:建立了特异性强、灵敏度良好的检测HIV-1 gp41抗原的双抗体夹心ELISA法.

  9. 牛病毒性腹泻病毒双抗体夹心ELISA检测方法的建立%Detection of bovine viral diarrhea virus by sandwich ELISA

    Institute of Scientific and Technical Information of China (English)

    高存福; 秦建华; 赵月兰; 包永占; 张宁

    2005-01-01

    将牛病毒性腹泻病毒超免疫血清以常规方法提取IgG,采用过碘酸钠法标记辣根过氧化物酶(HRP),建立了从粪样中检测牛病毒性腹泻病毒抗原的双抗体夹心ELISA.结果,抗体的最佳包被量为150μg/mL,酶标抗体最适工作浓度为1∶200;封闭液为50mL/L的兔血清;待检粪样及酶标抗体的感作时间为37℃ 120min;底物显色时间为室温15min.应用建立的检测方法对河北省8个大中型奶牛场298份乳牛腹泻粪样进行了检测,结果,阳性检出率为42.6%.

  10. Establishment and preliminary application of an ELISA method for determination of pertactin%检测百日咳黏着素的酶联免疫吸附法的建立及初步应用

    Institute of Scientific and Technical Information of China (English)

    王玲; 潘殊男; 张霖阳; 夏菊; 王宇星; 张萍; 肖詹蓉

    2014-01-01

    目的 建立定量检测白喉-破伤风-无细胞百日咳疫苗生产过程中黏着素(pertactin,Prn)的方法.方法 纯化的Prn免疫家兔以制备高效价血清抗Prn抗体.辛酸-硫酸铵沉淀法纯化抗Prn多克隆抗体并进行辣根过氧化物酶标记,建立双抗体夹心ELISA法.结果 建立的方法与丝状血凝素和百日咳毒素无交叉反应,特异性较好.该ELISA法在1.25 ~ 80.00μg/L Prn测量区间有最佳线性,相关系数>0.99.实验内和实验间检测64.0、32.0、16.0 μg/L Prn,变异系数为2.6%~7.7%,回收率为84.9%~ 95.5%,精密度和准确度均符合常规质控要求,因此该法的定量限度为16.0 μg/L.结论 建立的ELISA法可有效检测百日咳疫苗纯化过程中的Prn含量,为组分百日咳疫苗质量控制奠定了重要基础.%Objective To establish an ELISA method for quantitative determination of pertactin (Prn) during production of combined diphtheria,tetanus and acellular pertussis vaccine.Methods Rabbits were immuned with purified Prn to prepare high-titer serum antibodies against Prn.The polyclonal antibodies against Prn were purified by octanoic acid-ammonium sulfate precipitation method and labeled with horseradish peroxidase to establish a double antibody sandwich ELISA method.Results The ELISA method had good specificity for Prn and no cross reaction with filamentous hemagglutinin and pertussis toxin.The best linearity of the ELISA method was formed in the range of 1.25-80.00 μg/L (r > 0.99).The coefficient of variation and recover rates of intra-and inter-assay were 2.6%-7.7% and 84.9%-95.5% respectively when 64.0,32.0 and 16.0 μg/L standard Prn were detected,and the precision and accuracy both met requirements of quality control.The quantitative limit was 16.0 μg/L.Conclusion The established ELISA method can be applied to detecting Prn during purification of pertussis vaccine and lay the foundation for quality control of Prn.

  11. Enrichment-ELISA for Detection of Salmonella typhi From Food and Water Samples

    Institute of Scientific and Technical Information of China (English)

    S.KUMAR; K.BALAKRISHNA; HV.BATRA

    2008-01-01

    Objective Development of monoclonal antibody based sandwich enzyme linked immunosorbant assay(sELISA)for rapid detection of Salmonella enterica serovar typhi(S.typhi) from food and water samples and optimization of enrichment procedures for use with the developed sELISA to increase the detection Sensitivity of the assay. Methods Spleen cells from BALB/c mice immunized with flagellin(H=d)antigen of S.typhi were fused with Sp2/0 myeloma cells.The hybridoma cell line specific to H=d antigen was established.characterized and ascites raised against one of these clones.The hypefimmune serum to flagellin antigen was raised in New Zealand White rabbits.An sELISA was developed using polyclonalantibody as capture and monoelonal antibody as detection antibody.To design the efficient culture sUrategies for use with the sELISA.different pre-enrichment and enrichment brothswere evaluated.The mediaincluded buffered peptonewater(BPW)and brain heart infusion broth for pre-enrichment and selenite F broth and Rappaport-Vassiliadis broth as enrichment broths.The developed sELISA with preceding enrichment step in BPW(Enrichment-ELISA)was evaluated in various food samples artificially inoculated with S. typhi bacteria.Various food(30)and water(35)samples collected from field were also tested by Enrichment-ELISA and culture method. Results Out of four specific clones to H=d antigen,one clone(#2/56.IgG2a isotype)was usedin sELISA.The sELISA had the detectionlimit of 104-105 cfu of S.typhi.Of the various broths used with sELISA,BPW was found to yield maximum ELISA values.Enrichment-ELISA,when tested in artificially inoculated food samples,generally,could detect 102S.typhi cfu/mL within 10 h from variousfood rinses(meat,vegetable)and milk samples.After overnight enrichment in BPW,as less as 2 bacteria per 10 mL of milk,meat rinse.and chicken rinse could be detected.Only one of the field samples(water)gave false positive result by Enrichment-ELISA.Conclusion In comparison to culture

  12. Development and evaluation of an antibody ELISA for sarcoptic mange in sheep and a comparison with the skin-scraping method.

    Science.gov (United States)

    Rodríguez-Cadenas, F; Carbajal-González, M T; Fregeneda-Grandes, J M; Aller-Gancedo, J M; Huntley, J F; Rojo-Vázquez, F A

    2010-08-01

    In this work an indirect ELISA for detecting serum-specific IgG antibodies in sheep was developed using a crude saline extract from Sarcoptes scabiei var. ovis mites and then the repeatability of the ELISA outcomes was estimated. Subsequently, its diagnostic accuracy was evaluated by Receiver Operating Characteristics (ROC) analysis using a sample collected from the entire sheep population of western Castile and Leon region in Spain, and then compared with that of the skin-scraping method. The reference method used was a combination of clinical examination, skin-scraping analysis and epidemiological surveys, but it introduced selection and probably information biases. Furthermore, we attempted to identify biological factors useful to predict the sensitivity or specificity of the ELISA as determined by comparison with the reference method. Additionally, conventional latent-class analysis [Hui, S.L., Walter, S.D., 1980. Estimating the error rates of diagnostic tests. Biometrics 36, 167-171] was also used to estimate accuracy parameters. The between-run coefficient of variation (CV) for a standard serum was 8.8% and the within-run CV 4.3%. No significant deviation between the OD% means and strength positive correlation between the OD% values (r=0.98) were found for the results from two different batches of antigen. When compared to the reference method, the Area Under the ROC curve (AUC) for the reference population was 0.967 (95% CI: 0.949-0.985) for the ELISA and 0.915 (95% CI: 0.863-0.968) for the skin-scraping method. By logistic regression analysis, one explanatory biological factor-result to the skin-scraping method-and four explanatory biological factors-Tyroglyphidae individual status, Trichophyton verrucosum individual status, Oestrus ovis status of the flock and presence of adjacent animals with a clinical disease neighbour to S. scabiei infection-were found for diagnostic sensitivity and specificity of the ELISA, respectively, although this depended on the

  13. Separation and determination of trace uranium using a double-receptor sandwich supramolecule method based on immobilized salophen and fluorescence labeled oligonucleotide

    Energy Technology Data Exchange (ETDEWEB)

    Wu Minlong [College of Chemistry and Chemical Engineering, University of South China, Hengyang, Hunan 421001 (China); Liao Lifu, E-mail: lf_liao@yahoo.com.cn [College of Chemistry and Chemical Engineering, University of South China, Hengyang, Hunan 421001 (China); Zhao Minmin; Lin Yingwu; Xiao Xilin; Nie Changming [College of Chemistry and Chemical Engineering, University of South China, Hengyang, Hunan 421001 (China)

    2012-06-04

    Highlights: Black-Right-Pointing-Pointer We report a double-receptor sandwich method for separating and determining uranium. Black-Right-Pointing-Pointer One receptor used for separating uranium is an immobilized salophen. Black-Right-Pointing-Pointer Another used for determining uranium is a fluorescence labeled oligonucleotide. Black-Right-Pointing-Pointer The method utilizes the formation of supramolecule oligonucleotide-uranyl-salophen. Black-Right-Pointing-Pointer It has the advantages of high selectivity, high sensitivity and good stability. - Abstract: A double-receptor sandwich supramolecule method for the separation and determination of trace uranium was proposed in this paper. One receptor is a salophen which can react with uranyl to form a uranyl-salophen complex, and another receptor is an oligonucleotide which can bind uranyl to form oligonucleotide-uranyl-salophen supramolecule. The salophen was immobilized on the surface of silica gel particles and used as the solid phase receptor for separating uranium from solution. The oligonucleotide was labeled with a fluorescent group and used as the labeled receptor for quantitatively analyzing uranium. In the procedure of separation and determination, uranyl ion was first combined with the solid phase receptor and then conjugated with the labeled receptor to form the sandwich-type supramolecule. The labeled receptor in the sandwich supramolecule was then eluted and determined by fluorescence analysis. The experimental results demonstrate that this method has a number of advantages such as high selectivity, excellent pre-concentration capability, high sensitivity, good stability and low cost. Under optimal conditions, the linear range for the detection of uranium is 0.5-30.0 ng mL{sup -1} with a detection limit of 0.2 ng mL{sup -1}. The proposed method was successfully applied for the separation and determination of uranium in real samples with the recoveries of 95.0-105.5%.

  14. Fatigue characterization of Poly Vinyl Chloride (PVC) foam core sandwich composite using the G-control method

    DEFF Research Database (Denmark)

    Manca, Marcello; Berggreen, Christian; Carlsson, Leif A.

    2016-01-01

    This paper presents experimental results from cyclic crack propagation tests performed on sandwich specimens with glass/epoxy face sheets and Poly Vinyl Chloride (PVC) foam cores using the G-controlled cyclic energy release rate (ΔG) test procedure. The face material was tested in tension......, compression and shear to determine in-plane and out-of-plane mechanical properties, such as Young’s modulus, Poisson’s ratio and shear modulus. These properties were then used in an analytical model of the mixed-mode bending sandwich specimen to calculate compliance and energy release rate. Finite element...... analysis was used to determine the mode-mixity of the crack loading. Experimental crack growth cyclic tests were carried out on pre-cracked mixed-mode bending sandwich specimens with H45, H100 and H160 PVC foam cores under two mode-mixities (mode I and mode II dominant). Post-mortem analysis was performed...

  15. [Optimization of ELISA and immunoblot methods for the detection of IgG antibodies against old world hantaviruses in wild rodents].

    Science.gov (United States)

    Polat, Ceylan; Karataş, Ahmet; Sözen, Mustafa; Matur, Ferhat; Abacıoğlu, Hakan; Öktem, Mehmet Ali

    2016-04-01

    Hantaviruses infect humans via inhalation of viral particles in infected rodents' secretions such as saliva, urine and faeces or via direct contact with infected rodents. The rodent species that are known as the carriers of Dobrava (DOBV), Puumala (PUUV), Saaremaa (SAAV), Tula (TULV) and Seoul (SEOV) viruses are found in our country. The presence of specific antibodies against hantaviruses have been demonstrated in rodents collected from Black Sea and Aegean Regions of Turkey in 2004 for the first time. The first hantavirus-related hemorrhagic fever with renal syndrome (HFRS) cases were reported in Black Sea region in 2009. The determination of the hantavirus prevalence in wild life and rodent populations in the field is crucial for the information about hantavirus-related cases and to clarify the state of risk. There is no commercial product optimized for the screening of rodent serum samples in terms of HFRS agents like DOBV and PUUV that are widely seen in Eurasia as well as Turkey. In this study, the antigens belonging to the commercial enzyme-linked immunoassay (ELISA) and immunoblot tests that are produced for the screening of human sera were used for the development of antibody screening tests against hantavirus in rodent sera and were optimized. The most appropriate serum and conjugate dilutions were determined for the optimization of ELISA (Anti-Hantavirus Pool ELISA; Euroimmun, Germany) and immunoblot (Euroline Anti-Hanta Profile 1 strips; Euroimmun, Germany) methods. Optimized ELISA method was used for the screening and optimized immunoblot method was used for the confirmation. A total of 84 wild rodent sera that belonged to Apodemus and Microtus species were evaluated with this procedure and the cut-off value, sensitivity and specificity of optimized ELISA method were determined. For the optimization of ELISA 1/50, 1/100 and 1/200 serum dilutions and 1/10.000, 1/20.000 and 1/40.000 conjugate dilutions were tested. For the optimization of immunoblot, 1

  16. Sandwich教学法在医学寄生虫学教学中的体会%Experience of sandwich teaching method in teaching of medical parasitology

    Institute of Scientific and Technical Information of China (English)

    战廷正; 石焕焕; 刘登宇

    2013-01-01

    为引导学生自主学习,将Sandwich教学法引入医学寄生虫学教学.Sandwich教学法通过老师与学生、学生与学生之间的不断交流沟通,使学生发挥主观能动性,从而达到自主学习的目的.笔者将这种方法用于《医学寄生虫学》教学中,既能有效引导学生主动学习,又能培养学生分析问题和解决问题的能力.%To guide the independent learning of students,sandwich teaching method was introduced into teaching of medical parasitology.Teacher and students or students and students communicate with each other through sandwich teaching method,thus exploring the activeness of students and achieving the purpose of autonomous learning.Sandwich teaching method in teaching of medical parasitology can guide the students to active learning effectively as well as to cultivate students' ability for analyzing and solving problems.

  17. Sandwich DIY

    Institute of Scientific and Technical Information of China (English)

    肖蕾

    2006-01-01

    我们都知道sandwich是一种方便食品,就是在两片面包中加上一些肉和蔬菜。Sandwich这个名字来源于英国的一位桑威治伯爵(Earl of Sandwich)。据说这位伯爵嗜赌如命,就是到吃饭的时候也不愿停下来。于是他就叫侍者把肉、蛋、菜夹在面包片中,让他拿在手上边赌边吃。后来人们就把这种夹馅面包叫做sandwich。现在sandwich已成为风靡世界的快餐食品(snack)了。Sandwich的做法其实很简单。如果你有两片面包,你几乎可以在这两片面包之间夹上任何食物来给自己做一个三明治。下面就让我们试一试,做一个三明治来吃。第一步:在一片面包上抹上黄油(butter)或植物黄油,在另一片面包上抹上蛋黄酱(mayonnaise)和芥末酱(mustard)。喜欢吃番茄酱(catsup)也可以放番茄酱!第二步:把花生酱(peanut butter)或者乳酪片(cream)、熟肉片放在涂了黄油的面包片上。想吃什么肉就放什么肉,香肠也可以!第三步:在乳酪上面放酸黄瓜片、番茄片和生菜。也可以根据个人的口味再放些乳酪、芥末酱和(或)番茄酱、洋葱、辣椒、盐、黑胡椒和醋。第四步:将第二片面包盖在上面,就做成了一个sand...

  18. Standard Test Method for Water Absorption of Core Materials for Structural Sandwich Constructions

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    2001-01-01

    1.1 This test method covers the determination of the relative amount of water absorption by various types of structural core materials when immersed or in a high relative humidity environment. This test method is intended to apply to only structural core materials; honeycomb, foam, and balsa wood. 1.2 The values stated in SI units are to be regarded as the standard. The inch-pound units given may be approximate. This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.

  19. 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.

  20. Characterization of the MoO3/Ag grids/MoO3 sandwich electrode deposited on flexible substrate via thermal deposition method

    Science.gov (United States)

    Wang, Chen-Tao; Ting, Chu-Chi; Li, Shan-Rong; Chu, Sheng-Yuan

    2016-09-01

    In this paper, we will discuss the characteristics of the flexible sandwich electrode. We fabricate the MoO3/Ag grids/MoO3 via thermal deposition method. We will measure the bending test and the optical and electric characteristics. The conclusion of the MoO3/Ag grids/MoO3 will compare with the MoO3/Ag film/MoO3 and ITO flexible electrodes. This sandwich electrode will increase the transmittance by less silver coverage but the MoO3/Ag grids/MoO3 have lower sheet resistance compared with MoO3/Ag film/MoO3. Therefore, we propose this new electrode structure is proper for application of OLEDs.

  1. Analysis of ELISA method in the diagnosis of treponema pallidum infection%诊断梅毒螺旋体感染中ELISA法的检验分析

    Institute of Scientific and Technical Information of China (English)

    丁兆明

    2016-01-01

    目的 探讨诊断梅毒螺旋体感染中ELISA法的检验分析效果.方法 收集我院2014年8月至2015年8月入院的70例接受疑似梅毒螺旋体感染治疗的患者,分别对所有患者采用梅毒螺旋体ELISA试验(ELISA法)及快速血浆反应素试验(RPR法)进行检测,比较两种方法的检测结果以及诊断效能等.结果 临床确诊梅毒患者50例,非患者20例;患者RPR法检测阳性43例(61.4%),检测阴性27例(38.6%);患者ELISA法检测阳性47例(67.1%),检测阴性23例(32.9%);ELISA法灵敏度90.0%,漏诊率10.0%,特异度93.3%,误诊率6.7%,正确指数83.3%;RPR法灵敏度80.0%,漏诊率20.0%,特异度85.0%,误诊率15.0%,正确指数65.0%;ELISA法的灵敏度、特异度及正确指数均高于RPR法;ELISA法的漏诊率及误诊率均低于RPR法.结论 诊断梅毒螺旋体感染中ELISA法的检验分析效果显著,具有临床诊断借鉴意义.%Objective To investigate the effect of ELISA method in the diagnosis of treponema pallidum infection.Methods 70 patients with suspected treponema pallidum infection treated in our hospital from August 2014 to August 2015 were collected,all patients underwent treponema pallidum ELISA assay (ELISA method) and rapid plasma reagin (RPR method) testing.Test results and diagnostic performance of two methods were compared.Results There were 50 patients with clinical diagnosis of syphilis,20 cases of non-patient.There were 43 positive cases (61.4%) detected by RPR,27 negative cases (38.6%);there were 47 positive cases (67.1%) detected by ELISA,23 negative cases (32.9%).The sensitivity of ELISA method was 90.0%,missed diagnosis rate was 10.0%,specificity was 93.3%,misdiagnosis rate was 6.7%,correct index was 83.3%;the sensitivity of RPR method was 80.0%,missed diagnosis rate was 20.0%,specificity was 85.0%,misdiagnosis rate was 15.0%,correct index was 65.0%;sensitivity,specificity and correct index of ELISA method were

  2. Nanospherical Brush as Catalase Container for Enhancing the Detection Sensitivity of Competitive Plasmonic ELISA.

    Science.gov (United States)

    Huang, Xiaolin; Chen, Rui; Xu, Hengyi; Lai, Weihua; Xiong, Yonghua

    2016-02-01

    Plasmonic enzyme-linked immunosorbent assay (pELISA) based on catalase (CAT)-mediated gold nanoparticle growth shows great potential for the determination of disease-related biomarkers at ultralow concentrations by using sandwich formats. However, the relatively low sensitivity of this strategy using competitive formats limits its adoption for hapten detection. Herein, we present an improved competitive pELISA for ultrasensitive detection of ochratoxin A (OTA), where silica nanoparticles carrying poly(acrylic acid) brushes (SiO2@PAA) were used to decrease the affinity of competing antigens to anti-OTA monoclonal antibodies and amplify the signal as a "CAT container" (SiO2@PAA@CAT). The developed competitive pELISA exhibits extremely high sensitivity for OTA with detection limits of 10(-18) and 5 × 10(-20) g/mL by the naked eye and microplate reader, respectively. These values are at least 7 orders of magnitude lower than that of competitive CAT-based pELISA (10(-11) g/mL by the naked eye) and 8 orders of magnitude lower than that of horseradish peroxidase-based conventional ELISA (10(-11) g/mL by the microplate reader), respectively. Reliability and robustness of the proposed method were evaluated using actual agricultural products and human serum samples. This study demonstrated the potential of this modified method in practical applications involving the ultrasensitive detection of mycotoxins or other haptens.

  3. Development of a species-specific coproantigen ELISA for human Taenia solium taeniasis.

    Science.gov (United States)

    Guezala, Maria-Claudia; Rodriguez, Silvia; Zamora, Humberto; Garcia, Hector H; Gonzalez, Armando E; Tembo, Alice; Allan, James C; Craig, Philip S

    2009-09-01

    Taenia solium causes human neurocysticercosis and is endemic in underdeveloped countries where backyard pig keeping is common. Microscopic fecal diagnostic methods for human T. solium taeniasis are not very sensitive, and Taenia saginata and Taenia solium eggs are indistinguishable under the light microscope. Coproantigen (CoAg) ELISA methods are very sensitive, but currently only genus (Taenia) specific. This paper describes the development of a highly species-specific coproantigen ELISA test to detect T. solium intestinal taeniasis. Sensitivity was maintained using a capture antibody of rabbit IgG against T. solium adult whole worm somatic extract, whereas species specificity was achieved by utilization of an enzyme-conjugated rabbit IgG against T. solium adult excretory-secretory (ES) antigen. A known panel of positive and negative human fecal samples was tested with this hybrid sandwich ELISA. The ELISA test gave 100% specificity and 96.4% sensitivity for T. solium tapeworm carriers (N = 28), with a J index of 0.96. This simple ELISA incorporating anti-adult somatic and anti-adult ES antibodies provides the first potentially species-specific coproantigen test for human T. solium taeniasis.

  4. Aptamer-based Sandwich Assay and its Clinical Outlooks for Detecting Lipocalin-2 in Hepatocellular Carcinoma (HCC)

    Science.gov (United States)

    Lee, Kyeong-Ah; Ahn, Ji-Young; Lee, Sang-Hee; Singh Sekhon, Simranjeet; Kim, Dae-Ghon; Min, Jiho; Kim, Yang-Hoon

    2015-01-01

    We validated a single-stranded, DNA aptamer-based, diagnostic method capable of detecting Lipocalin-2 (LCN2), a biomarker from clinically relevant hepatocellular carcinoma (HCC) patient serum, in the sandwich assay format. Nine aptamers (LCN2_apta1 to LCN2_apta9) for LCN2 were screened with SELEX processes, and a sandwich pair (LCN2_apta2 and LCN2_apta4) was finally chosen using surface plasmon resonance (SPR) and dot blotting analysis. The result of the proposed aptamer sandwich construction shows that LCN2 was sensitively detected in the concentration range of 2.5–500 ng mL−1 with a limit of detection of 0.6 ng mL−1. Quantitative measurement tests in HCC patients were run on straight serum and were compared with the performance of the conventional antibody-based ELISA kit. The aptamer sandwich assay demonstrated an excellent dynamic range for LCN2 at clinically relevant serum levels, covering sub-nanogram per mL concentrations. The new approach offers a simple and robust method for detecting serum biomarkers that have low and moderate abundance. It consists of functionalization, hybridization and signal read-out, and no dilution is required. The results of the study demonstrate the capability of the aptamer sandwich assay platform for diagnosing HCC and its potential applicability to the point-of-care testing (POCT) system. PMID:26039737

  5. Development and Evaluation of Stitched Sandwich Panels

    Science.gov (United States)

    Stanley, Larry E.; Adams, Daniel O.; Reeder, James R. (Technical Monitor)

    2001-01-01

    This study explored the feasibility and potential benefits provided by the addition of through-the-thickness reinforcement to sandwich structures. Through-the-thickness stitching is proposed to increase the interlaminar strength and damage tolerance of composite sandwich structures. A low-cost, out-of-autoclave processing method was developed to produce composite sandwich panels with carbon fiber face sheets, a closed-cell foam core, and through-the-thickness Kevlar stitching. The sandwich panels were stitched in a dry preform state, vacuum bagged, and infiltrated using Vacuum Assisted Resin Transfer Molding (VARTM) processing. For comparison purposes, unstitched sandwich panels were produced using the same materials and manufacturing methodology. Test panels were produced initially at the University of Utah and later at NASA Langley Research Center. Four types of mechanical tests were performed: flexural testing, flatwise tensile testing, core shear testing, and edgewise compression testing. Drop-weight impact testing followed by specimen sectioning was performed to characterize the damage resistance of stitched sandwich panels. Compression after impact (CAI) testing was performed to evaluate the damage tolerance of the sandwich panels. Results show significant increases in the flexural stiffness and strength, out-of-plane tensile strength, core shear strength, edgewise compression strength, and compression-after-impact strength of stitched sandwich structures.

  6. Specific Recognition of Influenza A/H1N1/2009 Antibodies in Human Serum: A Simple Virus-Free ELISA Method

    Science.gov (United States)

    Alvarez, Mario M.; López-Pacheco, Felipe; Aguilar-Yañez, José M.; Portillo-Lara, Roberto; Mendoza-Ochoa, Gonzalo I.; García-Echauri, Sergio; Freiden, Pamela; Schultz-Cherry, Stacey; Zertuche-Guerra, Manuel I.; Bulnes-Abundis, David; Salgado-Gallegos, Johari; Elizondo-Montemayor, Leticia; Hernández-Torre, Martín

    2010-01-01

    Background Although it has been estimated that pandemic Influenza A H1N1/2009 has infected millions of people from April to October 2009, a more precise figure requires a worldwide large-scale diagnosis of the presence of Influenza A/H1N1/2009 antibodies within the population. Assays typically used to estimate antibody titers (hemagglutination inhibition and microneutralization) would require the use of the virus, which would seriously limit broad implementation. Methodology/Principal Findings An ELISA method to evaluate the presence and relative concentration of specific Influenza A/H1N1/2009 antibodies in human serum samples is presented. The method is based on the use of a histidine-tagged recombinant fragment of the globular region of the hemagglutinin (HA) of the Influenza A H1N1/2009 virus expressed in E. coli. Conclusions/Significance The ELISA method consistently discerns between Inf A H1N1 infected and non-infected subjects, particularly after the third week of infection/exposure. Since it does not require the use of viral particles, it can be easily and quickly implemented in any basic laboratory. In addition, in a scenario of insufficient vaccine availability, the use of this ELISA could be useful to determine if a person has some level of specific antibodies against the virus and presumably at least partial protection. PMID:20418957

  7. Specific recognition of influenza A/H1N1/2009 antibodies in human serum: a simple virus-free ELISA method.

    Directory of Open Access Journals (Sweden)

    Mario M Alvarez

    Full Text Available BACKGROUND: Although it has been estimated that pandemic Influenza A H1N1/2009 has infected millions of people from April to October 2009, a more precise figure requires a worldwide large-scale diagnosis of the presence of Influenza A/H1N1/2009 antibodies within the population. Assays typically used to estimate antibody titers (hemagglutination inhibition and microneutralization would require the use of the virus, which would seriously limit broad implementation. METHODOLOGY/PRINCIPAL FINDINGS: An ELISA method to evaluate the presence and relative concentration of specific Influenza A/H1N1/2009 antibodies in human serum samples is presented. The method is based on the use of a histidine-tagged recombinant fragment of the globular region of the hemagglutinin (HA of the Influenza A H1N1/2009 virus expressed in E. coli. CONCLUSIONS/SIGNIFICANCE: The ELISA method consistently discerns between Inf A H1N1 infected and non-infected subjects, particularly after the third week of infection/exposure. Since it does not require the use of viral particles, it can be easily and quickly implemented in any basic laboratory. In addition, in a scenario of insufficient vaccine availability, the use of this ELISA could be useful to determine if a person has some level of specific antibodies against the virus and presumably at least partial protection.

  8. The sandwich sign

    Directory of Open Access Journals (Sweden)

    Nasreen Mahomed

    2012-09-01

    Full Text Available The sandwich sign refers to the sandwiching of mesenteric vessels and fat by enlarged mesenteric nodes on cross-sectional imaging, commonly occurring in lymphoma, but not specific to lymphoma. The sign is radiologically indistinguishable from post-transplant lymphoproliferative disorders. The radiological significance of the sandwich sign is in suggesting the diagnosis of lymphoma so that appropriate treatment may be initiated early as the tumour has a rapid growth pattern.

  9. Quantitative analysis of four protein biomarkers: An automated microfluidic cartridge-based method and its comparison to colorimetric ELISA.

    Science.gov (United States)

    Dysinger, Mark; Marusov, Greg; Fraser, Stephanie

    2017-09-13

    Biomarker quantitation with ligand binding assays has matured greatly in recent years. This maturation has been partly in response to demands for more data points from fewer samples or less available sample volume. Multiplexing offers opportunities to acquire data for multiple analytes from single sample assay iterations, but has its own unique challenges and limitations. ProteinSimple has developed Simple Plex™, an automated immunoassay platform consisting of microfluidic cartridge-based assays run on the Ella instrument. Ella subverts traditional multiplexing challenges by rapidly performing triplicate measurements of up to four different analytes simultaneously, each in their own respective assay vessels and all from a single sample. Here we describe a comparison of the Simple Plex platform versus colorimetric ELISA and their respective abilities to quantitate four common biomarkers (MCP-1/CCL2, VEGF-A, TNF-α, and IL-6) from twenty-eight healthy individual donor plasma samples. Each biomarker was tested on the two platforms on each of two days. Ella analysis required significantly reduced sample volume, manual steps, and total time. Overall, Ella was able to quantify results for all twenty-eight samples for each of the four biomarkers. In contrast, ELISA was able to measure quantifiable results within respective calibration curve ranges for MCP-1/CCL2 (96% of samples) and VEGFA (7% of samples). For TNF-α and IL-6, ELISA was not sensitive enough to quantify any samples in the assay ranges. This stark difference in quantitative results underscores Ella's ability to multiplex without compromising sensitivity, and has far reaching potential for biomarker panel measurement in support of diagnosis, prognosis, and monitoring of disease. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Predicting the cross-reactivities of polycyclic aromatic hydrocarbons in ELISA by regression analysis and CoMFA methods

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Yan-Feng; Dai, Shu-Gui [College of Environmental Science and Engineering, Nankai University, Key Laboratory for Pollution Process and Environmental Criteria of Ministry of Education, Tianjin (China); Ma, Yi [College of Chemistry, Nankai University, Institute of Elemento-Organic Chemistry, Tianjin (China); Gao, Zhi-Xian [Institute of Hygiene and Environmental Medicine, Tianjin (China)

    2010-07-15

    Immunoassays have been regarded as a possible alternative or supplement for measuring polycyclic aromatic hydrocarbons (PAHs) in the environment. Since there are too many potential cross-reactants for PAH immunoassays, it is difficult to determine all the cross-reactivities (CRs) by experimental tests. The relationship between CR and the physical-chemical properties of PAHs and related compounds was investigated using the CR data from a commercial enzyme-linked immunosorbent assay (ELISA) kit test. Two quantitative structure-activity relationship (QSAR) techniques, regression analysis and comparative molecular field analysis (CoMFA), were applied for predicting the CR of PAHs in this ELISA kit. Parabolic regression indicates that the CRs are significantly correlated with the logarithm of the partition coefficient for the octanol-water system (log K{sub ow}) (r{sup 2}=0.643, n=23, P<0.0001), suggesting that hydrophobic interactions play an important role in the antigen-antibody binding and the cross-reactions in this ELISA test. The CoMFA model obtained shows that the CRs of the PAHs are correlated with the 3D structure of the molecules (r{sub cv}{sup 2}=0.663, r{sup 2}=0.873, F{sub 4,32}=55.086). The contributions of the steric and electrostatic fields to CR were 40.4 and 59.6%, respectively. Both of the QSAR models satisfactorily predict the CR in this PAH immunoassay kit, and help in understanding the mechanisms of antigen-antibody interaction. (orig.)

  11. Detección de alérgenos de huevo en fideos secos por método de elisa Detection of egg allergens in dried pasta using elisa method

    Directory of Open Access Journals (Sweden)

    María Julieta Binaghi

    2012-06-01

    acuerdo con los resultados obtenidos el método ELISA permitió la detección de alergenos de huevo en varias de las muestras provistas por la industria y también en algunas muestras comerciales con leyendas precautorias. Para una correcta rotulación de estos productos resultaría necesario contar con valores umbrales establecidos por la Autoridad Sanitaria Nacional.As the mandatory declaration of allergens will be soon included in the Argentine Food Code (article 235, section seven, it is necessary to implement control methodologies to detect the possible existence of allergens in commercial products. The presence of egg allergens in pasta and in products with common wheat flour and /or durum wheat semolina is possible as a result of cross contamination. The objective of this work was to evaluate, using ELISA method, the possible contamination with egg proteins in products made with common wheat flour and /or durum wheat semolina provided by a manufacturer to verify a possible contamination with egg proteins and to analyse commercial products with precautionary labels. Nineteen products made with common wheat flour and /or durum wheat semolina provided by a manufacturer and nine commercial products (dry pasta were analysed. The egg allergen was analyzed using the r-biopharm kit - ELISA. The samples were analysed in duplicates following the kit protocol. Among the samples provided by manufacturers there were products produced in line with the production of pasta with egg, with values below the quantification limit of the kit; products produced with a low percentage of milled products that could be made with egg, in which very low quantities of egg allergens were found (lower than 5 ppm, and products produced with a high percentage of milled products that could be made with egg and in which higher concentrations were obtained (higher than 5 ppm; whole dried egg. Five of the commercial samples presented concentrations that were below or similar to the quantification limit

  12. Use of the Filter-Sandwich carriers in continuous effectiveness monitoring of slurry treatment methods as an element improving biosafety in agriculture.

    Science.gov (United States)

    Skowron, Krzysztof; Olszewska, Halina; Paluszak, Piotr; Skowron, Karolina Jadwiga; Bauza-Kaszewska, Justyna

    2013-01-01

    Slurry, due to high microbiological contamination, requires hygienization before spreading. The agricultural usage of treated slurry has to guarantee biosafety. Therefore, constant monitoring of the slurry treatment process should be conducted. The use of Filter-Sandwich carriers seems to be a prospective solution. The aim of the research was to test whether Filter-Sandwich carriers influence the survivability of microorganisms during the slurry hygienization process and hence, whether they are safe for the environment. Raw cattle and swine slurry with different dry matter content was the research material. Salmonella Senftenberg W775 rods were introduced directly into the slurry and into the carriers placed in the liquid excrements stored at 4 and 20ºC, and underwent anaerobic digestion at 35ºC. The number of tested bacteria obtained from the slurry and carriers was determined using the MPN method with proper microbiological media. The values of physicochemical parameters of the raw and treated slurry were determined, both for the carriers and for slurry only. Biosafety control was also conducted for the carriers in slurry containers. The differences in the theoretical survivability between Salmonella Senftenberg W775 re-isolated from the slurry and the carriers, and in the values of the selected physicochemical parameters obtained at the end of the process, were not statistically significant. The re-contamination of the sterile slurry caused by the bacteria in the carrier was not observed after placement of the carrier with inoculated material. The conducted research proves the usefulness of Filter-Sandwich carriers for continuous hygienization monitoring of the slurry treatment process. This refers not only to the semi-technical scale, but also to the full-scale process.

  13. Detection of Shiga toxin-producing Escherichia coli by sandwich enzyme-linked immunosorbent assay using chicken egg yolk IgY antibodies

    Directory of Open Access Journals (Sweden)

    Yanil R Parma

    2012-06-01

    Full Text Available Enterohemorrhagic Escherichia coli (EHEC, a subset of Shiga toxin producing E. coli (STEC is associated with a spectrum of diseases that includes diarrhea, hemorrhagic colitis and a life-threatening hemolytic uremic syndrome (HUS. Regardless of serotype, Shiga toxins (Stx1 and/or Stx2 are uniformly expressed by all EHEC, and so exploitable targets for laboratory diagnosis of these pathogens. In this study, a sandwich ELISA for determination of Shiga toxin (Stx was developed using anti-Stx2 B subunit antibodies and its performance was compared with that of the Vero cell assay and a commercial immunoassay kit. Chicken IgY was used as capture antibody and a HRP-conjugated rabbit IgG as the detection antibody. The anti-Stx2B IgY was harvested from eggs laid by hens immunized with a recombinant protein fragment. Several parameters were tested in order to optimize the sandwich ELISA assay, including concentration of antibodies, type and concentration of blocking agent, and incubation temperatures. Supernatants from 42 STEC strains of different serotypes and stx variants, including stx2EDL933, stx2vha, stx2vhb, stx2g, stx1EDL933 and stx1d were tested. All Stx variants were detected by the sandwich ELISA, with a detection limit of 400 ng /ml Stx2. Twenty three strains negative for stx genes, including different bacteria species, showed no activity in Vero cell assay and produced negative results in ELISA, except for 2 strains. Our results show that anti-Stx2B IgY sandwich ELISA could be used in routine diagnosis as a rapid, specific and economic method for detection of Shiga toxin-producing E. coli.

  14. THE SIGNIFICANCE ON USING GOLD STANDARD METHOD FOR REEXAMING THE HBSAG WEAK POSITIVE SAMPLES BY ELISA METHOD%金标法对ELISA法检测HBSAg弱阳性标本复查的意义

    Institute of Scientific and Technical Information of China (English)

    栾雪静

    2011-01-01

    [目的]总结分析金标法对ELISA法检测HBSAg弱阳性标本复查的重要性,进一步提高ELISA法手工检测的质量. [方法]以0.1≤A≤0.4作为弱阳性,从4 970例ELISA手工法检测HBSAg的结果中选出37例弱阳性标本用金标法进行复检. [结果]用ELISA法检测的37例弱阳性标本,用金标法复查,其中阴性7例,再次用ELISA法复查检出阳性4例. [结论] ELISA法联合金标法检测HBSAg可以提高检测结果的准确性.%[Objective] To analyze the importance of using the gold standard method to reexamine HBSAg weakly positive samples detected by ELISA method, to further improve the quality of ELISA method's manual testing. [Methods] 0.1 :SA ≤0.4 as weakly positive, selected 37 weak positive samples from the assay results of 4 970 cases of HBSAg weakly positive samples detected by the ELISA manual testing, and reexamined these 37 with gold standard method. [Results] Used Gold standard method to reexamine these 37 weak positive samples detected by ELISA, the result was that 7 cases showed be negative with review of gold standard method. Reexamined them with ELISA again, positive was in 4 cases. [Conclusion] Combining the gold standard method and ELISA method in detecting HBSAg can improve the accuracy of test results.

  15. Discussing Interior Quality Control Method of ELISA Qualitative Test%ELISA法定性试验室内质控方法探讨

    Institute of Scientific and Technical Information of China (English)

    丁海明; 潘婉仪

    2011-01-01

    Objective To discuss ELISA qualitative test inside quality control method. Methods Detected HBsAg,HBsAb,HBeAg,HBeAb and HBcAb of weakly positive oc matter and 900 patients serum by ELISA method,analyse QC data with Levey-Jennings and half hold range method. Results 25 patients serum in 900 patients serum appeared false-negative and false-positive alone utilize Levey-Jennings QC map. Conclusion Combine Levey-Jennings oc map and half hold range method wben utilize ELISA qualitative test interior quality control method,reasonable choose fit experimental QC method by experimentation purpose.%目的 探讨ELISA法定性试验室内质控方法.方法 以ELISA法检测弱阳性质控品和900份病人血清的HBsAg,HBsAb,HBeAg,HBeAb和HBcAb,用Levey-Jennings质控图法和半固定范围法分别分析质控数据.结果 单独应用Levey-Jennings质控图法时,900份血清中有25份标本阴阳性出现误判.结论 在应用ELISA法定性试验室内质控方法时,结合Levey-Jennings质控图法和半固定范围法,根据实验目的合理选择适合各实验的质控方法.

  16. ELISA 方法筛查 TORCH 感染的质量控制方法探讨%Study on quality control of ELISA method for screening TORCH infection

    Institute of Scientific and Technical Information of China (English)

    迟绍琴; 陈奕微; 师宏词; 孙宇飞; 郑立新

    2015-01-01

    目的:探讨实验室定性检测TORCH感染指标(风疹病毒IgG、巨细胞病毒IgG和IgM、弓形虫IgG和IgM ,简称五项)的室内质量控制方法。方法采用统计学方法,正态分布数据,用ELISA方法检测的阳性率比值和标准差,设定1+2s为失控规则,绘制半Lerey‐Jennings质控图;非正态分布数据、小概率事件,则采用直接概率计算法,回顾分析57批次的检测数据。结果风疹病毒IgG阳性率为86.66%、巨细胞病毒 IgG/IgM 的阳性率为98.87%和0.13%、弓形虫 IgG/IgM 阳性率为2.43%和1.71%,五项指标在临界值范围的分别有151、3、5、176、27个标本数据。失控次数为巨细胞病毒IgG 1次,弓形虫IgG/IgM 分别是1次和4次。结论 ELISA定性检测TORCH感染指标的室内质控,可以采用日常检测阳性率或阴性率数据监控假阳性。临界值范围的标本应当进一步复检或确认实验。%Objective Onto investigate the indoor quality control method for qualitatively detecting the laboratory indicators of TORCH infection (rubella virus IgG ,cytomegalovirus IgG and IgM ,toxoplasma IgG and IgM ) .Methods The statistical method , normal distribution data ,ratio and standard deviation of positive rate detected by the ELISA method were adopted ,1+2s was set as the out of control rules ,the semi Lerey‐Jennings quality control chart was drawn;the direct probability calculation method was a‐dopted for the non‐normal distribution data and small probability event .The testing data of 57 batches were retrospectively ana‐lyzed .Results The positive rate of rubella virus IgG was 86 .66% ,cytomegalovirus IgG/IgM positive rates were 98 .87% and 0 .13% ,toxoplasma gondii IgG/IgM positive rates were 2 .43% and 1 .71% ,the data of 151 ,3 ,5 ,176 ,27 samples had the critical value range of five indicators .The number of out of control was once for cytomegalovirus IgG ,once and 4 times for Toxoplasma

  17. Evaluation of measurement of human TNF in plasma by ELISA.

    Science.gov (United States)

    Engelberts, I; Möller, A; Schoen, G J; van der Linden, C J; Buurman, W A

    1991-04-01

    The performance of a sandwich-ELISA for TNF measurement in plasma and serum was studied. The ELISA was first statistically analyzed. Interassay coefficient of variance and the intraassay coefficient of variance for the concentration range between 0.5 and 5 ng/ml was less than 10%. The sensitivity of the sandwich-ELISA for TNF in culture medium was 10 pg/ml. The ELISA was shown to be specific for biologically active TNF, since a good correlation between the ELISA and the WEHI bioassay was observed when partially inactive, denatured TNF was measured. The effect of various anticoagulation systems on the reliability of human TNF measurement has been evaluated. The oxalate/NaF and EDTA systems were both appropriate, as appeared from the observed blockade of the production of TNF in the tube, either in the cell-glycolysis-blocked or in the calcium-depleted situation, respectively. An eventual decrease in the recovery of rTNF after collection of blood was prevented in the oxalate/NaF tubes. Recovery of TNF in the ELISA was diminished in the presence of plasma or serum. Techniques to enhance the efficiency of the measurement of TNF in plasma by ELISA were compared. The data indicate that in the presence of 1.1 M NaCl, the TNF masking effect of normal plasma was largely abrogated. The presence and role of inhibiting plasma components in plasma of healthy and diseased individuals are discussed.

  18. Development of ELISA-based methods to measure the anti-malarial drug chloroquine in plasma and in pharmaceutical formulations

    DEFF Research Database (Denmark)

    Khalil, Insaf F; Alifrangis, Michael; Recke, Camilla

    2011-01-01

    In Central and South America and Eastern and Southern Africa, Plasmodium vivax infections accounts for 71-81% and 5% of malaria cases, respectively. In these areas, chloroquine (CQ) remains the treatment of choice for P. vivax malaria. In addition, CQ has recently proven to be an effective HIV-1 ...... resistance. The aim of this study was to develop an inexpensive, simple antibody-based ELISA to measure CQ concentrations in tablets and in plasma.......In Central and South America and Eastern and Southern Africa, Plasmodium vivax infections accounts for 71-81% and 5% of malaria cases, respectively. In these areas, chloroquine (CQ) remains the treatment of choice for P. vivax malaria. In addition, CQ has recently proven to be an effective HIV-1...

  19. The current preference for the immuno-analytical ELISA method for quantitation of steroid hormones (endocrine disruptor compounds) in wastewater in South Africa.

    Science.gov (United States)

    Manickum, Thavrin; John, Wilson

    2015-07-01

    The availability of national test centers to offer a routine service for analysis and quantitation of some selected steroid hormones [natural estrogens (17-β-estradiol, E2; estrone, E1; estriol, E3), synthetic estrogen (17-α-ethinylestradiol, EE2), androgen (testosterone), and progestogen (progesterone)] in wastewater matrix was investigated; corresponding internationally used chemical- and immuno-analytical test methods were reviewed. The enzyme-linked immunosorbent assay (ELISA) (immuno-analytical technique) was also assessed for its suitability as a routine test method to quantitate the levels of these hormones at a sewage/wastewater treatment plant (WTP) (Darvill, Pietermaritzburg, South Africa), over a 2-year period. The method performance and other relevant characteristics of the immuno-analytical ELISA method were compared to the conventional chemical-analytical methodology, like gas/liquid chromatography-mass spectrometry (GC/LC-MS), and GC-LC/tandem mass spectrometry (MSMS), for quantitation of the steroid hormones in wastewater and environmental waters. The national immuno-analytical ELISA technique was found to be sensitive (LOQ 5 ng/L, LOD 0.2-5 ng/L), accurate (mean recovery 96%), precise (RSD 7-10%), and cost-effective for screening and quantitation of these steroid hormones in wastewater and environmental water matrix. A survey of the most current international literature indicates a fairly equal use of the LC-MS/MS, GC-MS/MS (chemical-analytical), and ELISA (immuno-analytical) test methods for screening and quantitation of the target steroid hormones in both water and wastewater matrix. Internationally, the observed sensitivity, based on LOQ (ng/L), for the steroid estrogens E1, E2, EE2, is, in decreasing order: LC-MSMS (0.08-9.54) > GC-MS (1) > ELISA (5) (chemical-analytical > immuno-analytical). At the national level, the routine, unoptimized chemical-analytical LC-MSMS method was found to lack the required sensitivity for meeting environmental

  20. Can a combination of the conformal thin-sandwich and puncture methods yield binary black hole solutions in quasi-equilibrium?

    CERN Document Server

    Hannam, M D; Cook, G B; Baumgarte, T W; Hannam, Mark D.; Evans, Charles R.; Cook, Gregory B.; Baumgarte, Thomas W.

    2003-01-01

    We consider combining two important methods for constructing quasi-equilibrium initial data for binary black holes: the conformal thin-sandwich formalism and the puncture method. The former seeks to enforce stationarity in the conformal three-metric and the latter attempts to avoid internal boundaries, like minimal surfaces or apparent horizons. We show that these two methods make partially conflicting requirements on the boundary conditions that determine the time slices. In particular, it does not seem possible to construct slices that are quasi-stationary and avoid physical singularities and simultaneously are connected by an everywhere positive lapse function, a condition which must obtain if internal boundaries are to be avoided. Some relaxation of these conflicting requirements may yield a soluble system, but some of the advantages that were sought in combining these approaches will be lost.

  1. Diagnosis of peste des petits ruminants infection in small ruminants through in-house developed Indirect ELISA: Practical considerations

    Directory of Open Access Journals (Sweden)

    K. K. Sharma

    2015-04-01

    Full Text Available Aim: The work was conducted to diagnose peste des petits ruminants (PPR outbreak through an in house developed indirect ELISA (thereafter referred as iELISA its comparison with other available diagnostic tests and description of practical considerations in its development, utility and limitations. Materials and Methods: An outbreak resembled to PPR occurred in two different places of southern Gujarat viz. Vapi and Navsari, affecting 622 animals, including both goat (n = 476 and sheep (n = 146. Animals displayed the typical signs of PPR at Vapi; however diarrhea was the inconsistent feature in animals of Navsari. The affection caused morbidity of 100% and mortality were 73.68% (n = 392/532 and 56.67% (n = 51/90 in Vapi and Navsari outbreaks, respectively. Relevant ante mortem and post mortem samples were collected from representative animals. At the outset of the epidemic no kit was available with us, so agar gel immunodiffusion (AGID was carried out and a commercial ELISA (cELISA kit was ordered for making diagnosis through antibody demonstration. Meanwhile, an iELISA was developed in house using PPR vaccine as antigen and protein G conjugated HRPO antibody as detector. Histopathology and results of sandwich ELISA were also used to diagnose PPR virus (PPRV in the outbreak. Results: The iELISA developed had detected PPRV antibodies in 22/24 samples (91.66%. Significant difference was observed in disease sensitivity pattern of two species by Chi-square test. While AGID failed to detect antibodies in any sample. Results were reconfirmed by comparing with commercially available cELISA kit. Conclusion: PPR is an economically important disease and for the rapid diagnosis of PPR the in house developed antibody capture iELISA can be a suitable cost effective alternative.

  2. Strongyloides stercoralis: a field-based survey of mothers and their preschool children using ELISA, Baermann and Koga plate methods reveals low endemicity in western Uganda.

    Science.gov (United States)

    Stothard, J R; Pleasant, J; Oguttu, D; Adriko, M; Galimaka, R; Ruggiana, A; Kazibwe, F; Kabatereine, N B

    2008-09-01

    To ascertain the current status of strongyloidiasis in mothers and their preschool children, a field-based survey was conducted in western Uganda using a combination of diagnostic methods: ELISA, Baermann concentration and Koga agar plate. The prevalence of other soil-transmitted helminthiasis and intestinal schistosomiasis were also determined. In total, 158 mothers and 143 children were examined from five villages within Kabale, Hoima and Masindi districts. In mothers and children, the general prevalence of strongyloidiasis inferred by ELISA was approximately 4% and approximately 2%, respectively. Using the Baermann concentration method, two parasitologically proven cases were encountered in an unrelated mother and child, both of whom were sero-negative for strongyloidiasis. No infections were detected by Koga agar plate method. The general level of awareness of strongyloidiasis was very poor ( < 5%) in comparison to schistosomiasis (51%) and ascariasis (36%). Strongyloidiasis is presently at insufficient levels to justify inclusion within a community treatment programme targeting maternal and child health. Better epidemiological screening is needed, however, especially identifying infections in HIV-positive women of childbearing age. In the rural clinic setting, further use of the Baermann concentration method would appear to be the most immediate and pragmatic option for disease diagnosis.

  3. Significance on using gold standard method for reexaming the HBSAg weak positive samples by ELISA method%金标法对ELISA法检测HBSAg弱阳性标本复查的意义

    Institute of Scientific and Technical Information of China (English)

    栾雪静

    2012-01-01

    目的 总结分析金标法对ELISA法检测HBSAg弱阳性标本复查的重要性,进一步提高ELISA法手工检测的质量.方法 以0.1≤A≤0.4作为弱阳性,从4 970例ELISA手工法检测HBSAg的结果中选出37例弱阳性标本用金标法进行复检.结果 用ELISA法检测的37例弱阳性标本,用金标法复查,其中阴性7例,再次用ELISA法复查检出阳性4例.结论 ELISA法联合金标法检测HBSAg可以提高检测结果的准确性.%OBJECTIVE To analyse the importance of using the gold standard method to reexamine HBSAg weakly positive samples detected by ELISA method, to further improve the quality of manual testing of ELISA method. METHODS 0.1 ≤A≤0.4 as weakly positive, selected 37 weak positive samples from the assay results of 4 970 cases of HBSAg weakly positive samples detected by the ELISA manual testing, and reexamined these 37 with gold standard method. RESULTS In 37 weak positive samples tested by ELISA method, 7 cases showed be negative when reexamined by gold standard method, reviewed with gold standard method, in which 4 cases were positive. CONCLUSION Combined gold standard method and ELISA method in detecting HBSAg can improve the accuracy of test results.

  4. b型流感嗜血杆菌多糖ELISA检测方法的建立%Development of an ELISA Method for Polyribosylribitolphosphate of Haemophilus influenzae Type b

    Institute of Scientific and Technical Information of China (English)

    祝婧烨; 沈坚; 秦洁; 马相虎; 王维; 瞿爱东

    2011-01-01

    Objective To develop an ELJSA method for polyribosylribitolphosphate (PRP) of Haemophilia infiuenzae type b (Hib). Methods Rabbits were immunized with inactivated Hib, 4 times a week for 6 weeks. Serum samples were collected, determined for antibody titer and subjected to immunoadsorption with non-specific antibody, from which rabbit anti-PRP antibody was purified. A double antibody sandwich ELISA method was developed by coating microtiter plate with the purified rabbit anti-PRP antibody, using HRP-labeled goat anti-rabbit IgC as the second antibody, then determined for optimal linear range of standard curve, and verified for specificity, accuracy and precision. Results The serum antibody titers of the immunized rabbits reached 1 : 2 800. The linear detection range and minimum detection limit of the developed method were 0. 030 - 0. 500 ng / ml and 0. 030 ng / ml respectively. All the detection results of protein in supernatant of E. coli, bovine serum albumin, LB medium and tetanus-Japanese encephalitis combined vaccine by the developed method were negative. The standard PRP at concentrations of 0. 500, 0. 250 and 0. 150 ng/ml were detected by the developed method, and the result showed that the intra-variation coefficient (CV) and recovery rate were 1. 16% ~ 2. 40% and 93. 2% ~ 107. 2%, while the inter-CV and recovery rate were 1. 57% ~ 4. 11% and 101. 3% ~ 101. 6%, respectively. Conclusion The developed ELISA method showed high specificity, accuracy and precision, which might be used for specific detection of PRP of Hib.%目的 建立b型流感嗜血杆菌(Haemophilus influenzae type b,Hib)多糖(Polyribosylribitolphosphate,PRP)的ELISA检测方法.方法 用灭活的Hib菌免疫家兔,1周免疫4次,共免疫6周,采血,检测血清抗体滴度.对兔免疫血清进行非特异性抗体免疫吸附,纯化兔抗PRP抗体,用纯化的兔抗PRP抗体包被酶标板,HRP标记的羊抗兔IgG作为二抗,建立双抗体夹心ELISA法检测PRP.确定标准曲线的

  5. Evaluation of biochemical and serological methods to identify and clustering yeast cells of oral Candida species by CHROMagar test, SDS-PAGE and ELISA

    Directory of Open Access Journals (Sweden)

    J. A. de O. Rodrigues

    Full Text Available The purpose of this work was to evaluate biochemical and serological methods to characterize and identify Candida species from the oral cavity. The strains used were five Candida species previously identified: C. albicans, C. guilliermondii, C. parapsilosis, C. krusei, C. tropicalis, and Kluyveromyces marxianus, as a negative control. The analyses were conducted through the SDS-PAGE associated with statistical analysis using software, chromogenic medium, and CHROMagar Candida (CA, as a differential medium for the isolation and presumptive identification of clinically important yeasts and an enzyme-linked immunoabsorbent assay (ELISA, using antisera produced against antigens from two C. albicans strains. This method enabled the screening of the three Candida species: C. albicans, C. tropicalis, and C. Krusei, with 100% of specificity. The ELISA using purified immunoglobulin G showed a high level of cross-reaction against protein extracts of Candida species. The SDS-PAGE method allowed the clustering of species-specific isolates using the Simple Matching coefficient, S SM = 1.0. The protein profile analysis by SDS-PAGE increases what is known about the taxonomic relationships among oral yeasts. This methodology showed good reproducibility and allows collection of useful information for numerical analysis on information relevant to clinical application, and epidemiological and systematical studies.

  6. ELISA u analitici hrane

    OpenAIRE

    Runje, Mislav; Cvrtila, Željka

    2006-01-01

    U radu su prikazane neke od mogućnosti korištenja “screening” testa ELISA u analitici namirnica. Utvrđivanje vrsta mesa u toplinski obrađenim ili, pak, toplinski neobrađenim proizvodima jedna je od primjena ELISA testova.

  7. Improved methods for urinary atrazine mercapturate analysis-Assessment of an enzyme-linked immunosorbent assay (ELISA) and a novel liquid chromatography-mass spectrometry (LC-MS) method utilizing online solid phase extraction (SPE)

    Energy Technology Data Exchange (ETDEWEB)

    Koivunen, Marja E. [Department of Entomology and the UC Davis Cancer Center, University of California, Davis (United States); Dettmer, Katja [Department of Entomology and the UC Davis Cancer Center, University of California, Davis (United States); Vermeulen, Roel [Division of Cancer Epidemiology and Genetics, National Cancer Institute, NIH, DHHS, Rockville, MD (United States); Bakke, Berit [Division of Cancer Epidemiology and Genetics, National Cancer Institute, NIH, DHHS, Rockville, MD (United States); National Institute of Occupational Health, Oslo (Norway); Gee, Shirley J. [Department of Entomology and the UC Davis Cancer Center, University of California, Davis (United States); Hammock, Bruce D. [Department of Entomology and the UC Davis Cancer Center, University of California, Davis (United States)]. E-mail: bdhammock@ucdavis.edu

    2006-07-21

    Elimination of interfering substances in urine by solid phase extraction (SPE) prior to analysis resulted in 10-fold improvement in the sensitivity of atrazine mercapturate (AM) enzyme-linked immunosorbent assay (ELISA) compared to previous reports. Of the two tested SPE systems, Oasis[reg] HLB and MCX, the mixed-mode MCX gave good recoveries (82%) of AM in spiked samples measured by ELISA, whereas the reverse-phase HLB phase was not compatible with the immunochemical method. At relatively high concentrations of urinary AM (>20 ng mL{sup -1}), sample dilution was effective enough for the elimination of interfering substances. The new liquid chromatography-mass spectrometry (LC-MS) method developed for AM utilizes online-SPE with Oasis[reg] HLB, column switching and a stable-isotope internal standard. The limit of quantification (0.05 ng mL{sup -1}) indicates improved sensitivity compared with most previously published LC-MS methods for AM. Validation of all three methods, LC-MS, ELISA + SPE and ELISA + dilution with spiked urine samples showed good correlation between the known and measured concentrations with R {sup 2} values of 0.996, 0.957 and 0.961, respectively. When a set (n = 70 plus 12 blind duplicates) of urine samples from farmers exposed to atrazine was analyzed, there was a good agreement (R {sup 2} = 0.917) between the log normalized data obtained by ELISA + SPE and LC-MS. High correlation among the data obtained by the two tested methods and the LC-MS method by the Center of Disease Control and Prevention (CDC), together with low variability among the blind duplicates, suggests that both methods reported here would be suitable for the analysis of urinary AM as a biomarker for human exposure of atrazine.

  8. Sandwich classification theorem

    Directory of Open Access Journals (Sweden)

    Alexey Stepanov

    2015-09-01

    Full Text Available The present note arises from the author's talk at the conference ``Ischia Group Theory 2014''. For subgroups FleN of a group G denote by Lat(F,N the set of all subgroups of N , containing F . Let D be a subgroup of G . In this note we study the lattice LL=Lat(D,G and the lattice LL ′ of subgroups of G , normalized by D . We say that LL satisfies sandwich classification theorem if LL splits into a disjoint union of sandwiches Lat(F,N G (F over all subgroups F such that the normal closure of D in F coincides with F . Here N G (F denotes the normalizer of F in G . A similar notion of sandwich classification is introduced for the lattice LL ′ . If D is perfect, i.,e. coincides with its commutator subgroup, then it turns out that sandwich classification theorem for LL and LL ′ are equivalent. We also show how to find basic subroup F of sandwiches for LL ′ and review sandwich classification theorems in algebraic groups over rings.

  9. A Colorimetric Enzyme-Linked Immunosorbent Assay (ELISA) Detection Platform for a Point-of-Care Dengue Detection System on a Lab-on-Compact-Disc.

    Science.gov (United States)

    Thiha, Aung; Ibrahim, Fatimah

    2015-05-18

    The enzyme-linked Immunosorbent Assay (ELISA) is the gold standard clinical diagnostic tool for the detection and quantification of protein biomarkers. However, conventional ELISA tests have drawbacks in their requirement of time, expensive equipment and expertise for operation. Hence, for the purpose of rapid, high throughput screening and point-of-care diagnosis, researchers are miniaturizing sandwich ELISA procedures on Lab-on-a-Chip and Lab-on-Compact Disc (LOCD) platforms. This paper presents a novel integrated device to detect and interpret the ELISA test results on a LOCD platform. The system applies absorption spectrophotometry to measure the absorbance (optical density) of the sample using a monochromatic light source and optical sensor. The device performs automated analysis of the results and presents absorbance values and diagnostic test results via a graphical display or via Bluetooth to a smartphone platform which also acts as controller of the device. The efficacy of the device was evaluated by performing dengue antibody IgG ELISA on 64 hospitalized patients suspected of dengue. The results demonstrate high accuracy of the device, with 95% sensitivity and 100% specificity in detection when compared with gold standard commercial ELISA microplate readers. This sensor platform represents a significant step towards establishing ELISA as a rapid, inexpensive and automatic testing method for the purpose of point-of-care-testing (POCT) in resource-limited settings.

  10. A Colorimetric Enzyme-Linked Immunosorbent Assay (ELISA Detection Platform for a Point-of-Care Dengue Detection System on a Lab-on-Compact-Disc

    Directory of Open Access Journals (Sweden)

    Aung Thiha

    2015-05-01

    Full Text Available The enzyme-linked Immunosorbent Assay (ELISA is the gold standard clinical diagnostic tool for the detection and quantification of protein biomarkers. However, conventional ELISA tests have drawbacks in their requirement of time, expensive equipment and expertise for operation. Hence, for the purpose of rapid, high throughput screening and point-of-care diagnosis, researchers are miniaturizing sandwich ELISA procedures on Lab-on-a-Chip and Lab-on-Compact Disc (LOCD platforms. This paper presents a novel integrated device to detect and interpret the ELISA test results on a LOCD platform. The system applies absorption spectrophotometry to measure the absorbance (optical density of the sample using a monochromatic light source and optical sensor. The device performs automated analysis of the results and presents absorbance values and diagnostic test results via a graphical display or via Bluetooth to a smartphone platform which also acts as controller of the device. The efficacy of the device was evaluated by performing dengue antibody IgG ELISA on 64 hospitalized patients suspected of dengue. The results demonstrate high accuracy of the device, with 95% sensitivity and 100% specificity in detection when compared with gold standard commercial ELISA microplate readers. This sensor platform represents a significant step towards establishing ELISA as a rapid, inexpensive and automatic testing method for the purpose of point-of-care-testing (POCT in resource-limited settings.

  11. 电流变夹层板滑模振动控制%Vibration Control of an Electrorheological Sandwich Plate Using Sliding Control Method

    Institute of Scientific and Technical Information of China (English)

    陈春强; 陈前

    2016-01-01

    为实现板结构振动控制,基于滑动模态控制思想,设计适用于电流变夹层结构的变刚度控制器,并且推导两类控制律——滑模控制律和简化的开关控制律。先将电流变液处理为可控黏弹性材料,基于Hamilton原理建立电流变夹层板有限元模型。设计滑模控制器时,首先将系统变换到模态空间,然后利用LQR最优控制理论设计控制面函数,设计电流变夹层板的滑模控制器,最后给出两个近似的半主动控制律。对悬臂电流变夹层板进行仿真分析,设计的滑模控制器能显著降低电流变夹层板振动水平,均方根降幅达到88.23%,取得明显优于被动控制的减振效果,体现电流变夹层结构在板结构振动控制应用中的前景。%Based on the sliding mode control algorithm, a new active variable stiffness controller was presented and employed to deduce both control laws:continuous sliding mode control law and ON-OFF sliding mode control law. First of all, assuming that the electrorheological (ER) fluids behave as viscoelastic materials and their storage modulus and loss modulus depend on the applied electric fields, the kinetic equations of a sandwich plate with the ER materials layer were derived based on the Hamilton’s principle and finite element method (FEM). The effects of the electric fields on the natural frequencies and modal loss factors of the sandwich plate were obtained. The kinetic equations of the sandwich plate system were transformed to a modal system before designing the sliding mode controller. Then, the sliding surface was designed by LQR optimal control theory, and two semi-active control laws were proposed. The results of numerical simulation of a cantilever ER sandwich plate show that this control method is very effective in attenuating the structural vibration. The mean square root of displacement at point P has depressed about 88%. The attenuation effect is much better than

  12. Development of ELISA-analysis methods for the quantification of bioactive natural products in plants, phytomedicines and in humans or similar.

    Science.gov (United States)

    Tanaka, H; Shoyama, Y

    1998-10-01

    In the course of a program in developing new ELISA-methods for the quantification of bioactive natural products in plants, phytomedicines and animals in a μg and ng scale, monoclonal antibodies against various natural products of medicinal and analytical importance have been developed. The ratio of hapten to bovine serum albumin (BSA) in an antigen conjugate was determined by matrix-assisted laser desorption/ionization (MALDI) of mass spectrometry. A hybridoma secreting monoclonal antibodies (MAb) was produced by fusing splenocytes immunized with an antigen-BSA conjugate with HAT-sensitive mouse myeloma cells. The cross-reaction of anti-forskolin antibodies with 7-deacetyl-forskolin was 5.6%. A very small cross-reaction appeared with other derivatives. The full measuring range of the assay extends from 5 ng to 5 μg/ml of forskolin. Immunoaffinity column chromatography using anti-forskolin MAbs appears to be far superior to previously published separation methods. The capacity of the immunoaffinity column as determined by ELISA is 9 μg/ml. Forskolin has been isolated directly from the crude extracts of tuberous roots and the callus culture of Coleus forskohlii. A MAb against Δ(1)-tetrahydrocannabinolic acid (THCA) was produced. The cross-reaction of anti-Δ(1)-THCA antibody against other cannabinoids was very wide. Many cannabinoids and a spiro-compound were reactive, but did not react with other phenolics. It became evident that this ELISA method was able to be applied to the biotransformation experiments of cannabinoids in plant tissue culture system. Anti-solamargine MAbs were produced. A method of determination for solasodine glycosides by using TLC-immunostaining was established. Solasodine glycosides separated by silica gel TLC were transferred to a polyvinylidene difluoride (PVDF) membrane. The membrane was treated with NaIO(4) solution followed by BSA, resulting in a solasodine glycoside-BSA conjugate. Immunostaining of solasodine glycosides was more

  13. Comparative Study of Three Different ELISA to Measure the Antibodies Against Infectious Bronchitis Virus in Vaccinated and Unvaccinated Broilers

    OpenAIRE

    Cardoso TC; Oliveira. C.; SEL Da-Silva; HL Ferreira; Pinto AA

    2001-01-01

    Broilers were spray-vaccinated (n=150) with H120 serotype at one-day-old, challenged after 28 days with M41 IBV serotype and after bled at day 28, 34 and 46 after challenged. The respective sera were tested by the indirect ELISA (I-ELISA), sandwich ELISA (S-ELISA), liquid phase blocking ELISA (LPB-ELISA) and the standard serum neutralization test (SNT). For this purpose, a total of 300 sera samples, 150 from non vaccinated and 150 from vaccinated broilers were titrated by all the serological ...

  14. Buckling Analysis of Debonded Sandwich Panel Under Compression

    Science.gov (United States)

    Sleight, David W.; Wang, John T.

    1995-01-01

    A sandwich panel with initial through-the-width debonds is analyzed to study the buckling of its faceskin when subject to an in-plane compressive load. The debonded faceskin is modeled as a beam on a Winkler elastic foundation in which the springs of the elastic foundation represent the sandwich foam. The Rayleigh-Ritz and finite-difference methods are used to predict the critical buckling load for various debond lengths and stiffnesses of the sandwich foam. The accuracy of the methods is assessed with a plane-strain finite-element analysis. Results indicate that the elastic foundation approach underpredicts buckling loads for sandwich panels with isotropic foam cores.

  15. Evaluation of a rabies ELISA as an alternative method to seroneutralisation tests in the context of international trade of domestic carnivores.

    Science.gov (United States)

    Wasniewski, M; Labbe, A; Tribout, L; Rieder, J; Labadie, A; Schereffer, J L; Cliquet, F

    2014-01-01

    For several years, international movements with pets have greatly increased. Most countries have relaxed their quarantine measures and adopted a scheme combining vaccination of pets against rabies followed by a serological test to check the efficacy of vaccination. This new scheme has been strongly supported by the OIE, WHO and the European Commission to facilitate the free movement of people and pets around the world. Currently, only two reference methods are recognised and prescribed (the FAVN test and the RFFIT) to measure rabies antibody levels in serum samples for international trade. They are reliable and valuable methods of assessing the efficacy of rabies vaccination but they are time-consuming and require well-trained people and specialised laboratory facilities. A few years ago, an ELISA (Platelia™ Rabies II kit ad usum Veterinarium) was developed for domestic carnivores and wildlife. To our knowledge, this ELISA is the only one certified and prescribed by the OIE. Following its marketing, one task of the EURL for rabies serology was to evaluate the performance of laboratories using this new kit. The results revealed that 26% of the participants, which were already approved laboratories for rabies serology, failed the inter-laboratory trial. Such unsatisfactory results have never been observed during any of the previous proficiency tests organised annually since 2000 by the EURL for rabies serology using reference methods. More investigations were undertaken through internal and collaborative studies to assess the performance of this newly marketed ELISA kit. The results of the internal study revealed that even with a specificity of 100%, the sensitivity evaluated on 593 samples of domestic carnivores came to 78.2%. An issue regarding the underestimation of serum titres was also revealed during the study. The results of a collaborative study involving 23 international laboratories reinforced the preliminary conclusions regarding lack of sensitivity

  16. Predicting safe sandwich production

    DEFF Research Database (Denmark)

    Birk, Tina; Duan, Zhi; Møller, Cleide Oliveira de Almeida

    2014-01-01

    and serving. However, Danish sandwich producing companies find it challenging to comply with this and have expressed a need for more flexibility. The Danish guidelines do allow for a prolongation of the acceptable time outside the cold chain, if the safety of the specific production can be documented....... There is, therefore, room for developing targeted tools for evaluating the time-temperature scenarios in sandwich production. This study describes a decision support tool developed to offer the producers more flexibility. Based on time/temperature measurements obtained during preparation combined......Time and temperature control is crucial to avoid growth of pathogens during production and serving of cold ready-to-eat meals. The Danish guidelines state that chilled foods, such as sandwiches, should not be outside the cold chain for more than 3 hours including the time for preparation...

  17. Influence of plywood grain direction on sandwich panel bending properties

    OpenAIRE

    Jaroslav Kljak; Mladen Brezović; Alan Antonović

    2009-01-01

    This paper investigates the influence of plywood grain direction on bending properties of a sandwich panel, as well as on stress distribution in each layer. Experimental sandwich panels (tnom= 29 mm) were made of two three-ply plywood panels and a rigid PVC core between them. Grain directions of plywood panels were between 0° and 90°, continuously raised by 15°. Seven models of sandwich panels were made. Bending properties of a sandwich panel was determined by three point bending method and s...

  18. Analytical determination of the ultimate strength of sandwich beams

    Science.gov (United States)

    Theotokoglou, Efstathios E.

    1996-09-01

    An analytical determination of the ultimate strength of a typical GRP/PVC sandwich beam has been performed. These beams represent common building practise in marine applications. Equations describing the behaviour of a sandwich panel under beam loading and various failure modes have been developed. The method has been applied to predict the ultimate load for a simple supported sandwich beam. The critical loads have been compared with those from the experimental investigation of a typical bulkhead-to-hull GRP/PVC sandwich T-joint under pull out forces.

  19. HBV infection treatment on using Lamivadine instructed by the method of ELISPOT, ELISA%ELISPOT、ELISA指导拉米夫定治疗HBV感染

    Institute of Scientific and Technical Information of China (English)

    施理; 林锋; 许小珍; 贾杰; 陈所贤; 周炎腾; 沈伟

    2013-01-01

    OBJECTIVE The immune response of the patients with HBV was compared by the method of Enzyme Link Immune Spot (ELISPOT) and Enzyme-Linked Immunosorbnent Assay (ELISA),which direct the usage of Lamivudine.METHODS Patients chronically infected with HBV were divided into patients in clerance period and patients in tolerance period by the method of ELISA and ELISPOT.The effect of Lamivudine in both groups were also studied.RESULTS For the patients in tolerance period and clerance period,the production of interferon-r (IFNr) from Peripheral blood mononuclear cells (PBMCs) by the stimulation of HBcAg were (578.37ng/L ± 7.73ng/L) and (655.25 ng/L ± 3.80ng/L) respectively,and 12.50% patients in tolerance period and 68.81% patients in clerance period were positive by the method of ELISA.By the method of ELISPOT,0% and 87.50% of patients in tolerance period and clerance period were positive respectively,and there was a statistical difference (P =4.23×10-8).There were statistical differences of HBV level (x20.01 (2)=15.13,P < 0.01),and reponses to Lamivudine (x20.01(2)=11.73,P< 0.01) between the patients in clerance period and in tolerance period.There was a statistical difference of HBV level in the patients with various responses to Lamivudine (x20.05 (4)=9.93,P < 0.05).CONCLUSION The response of the production of IFNr stimulated with HBcAg for the patients in clerance period is evidently different from those in tolerance period,and ELISPOT is better than ELISA.The effect of Lamivudine is related to the HBV level and immune response to HBV.%目的 酶联免疫斑点(enzyme link immunal spot,ELISPOT)、酶联免疫吸附试验(Enzyme-Linked Immunosorbnent Assay,ELISA)鉴别HBV感染免疫状态,指导拉米夫定治疗.方法 用ELISPOT及ELISA方法鉴别免疫耐受期、免疫清除期患者,研究2组对拉米夫定治疗效果.结果 用ELISA方法,2组患者PBMCs受HBcAg刺激分泌IFNr,分别为(578.37±7.73)ng/L、(655.25±3.80) ng

  20. On Sandwiched Singularities

    OpenAIRE

    Möhring, Konrad

    2004-01-01

    Sandwich-Singularitäten sind die Singularitäten auf derNormalisierung von Aufblasungen eines regulärenFlächenkeimes. In der Arbeit wird ein enger Zusammenhangzwischen Topologie und Deformationstheorie vonSandwich-Singularitäten einerseits und ebenenKurvensingularitäten andererseits dargestellt. NeueErgebnisse betreffen u.a. Deformationen vonnulldimensionalen komplexen Räumen in der Ebene, die durchvollständige Ideale beschrieben werden, z.B. wann'simultanes Aufblasen' der Fasern einer solchen...

  1. Use of a novel serum ELISA method and the tonsil-carrier state for evaluation of Mycoplasma hyosynoviae distributions in pig herds with or without clinical arthritis

    DEFF Research Database (Denmark)

    Nielsen, Elisabeth Okholm; Lauritsen, Klara Tølbøll; Friis, Niels Filskov

    2005-01-01

    determined by culture of cross-sectional samples of whole-blood (n = 238) and tonsil scrapings (n = 322), respectively. Levels of serum antibodies (n = 396) were measured by the novel indirect ELISA test. There was no significant difference in the ELISA results between the MhA and the MhC herds. Pigs...

  2. A modified DCB sandwich specimen for measuring mixed-mode cohesive laws

    DEFF Research Database (Denmark)

    Lundsgaard-Larsen, Christian; Sørensen, Bent F.; Berggreen, Christian;

    2008-01-01

    A test method is described for measuring cohesive laws for interfaces in sandwich structures. It is proposed to increase the bending stiffness of the sandwich faces by adhering steel bars onto the sandwich faces. This stiffening reduces rotations and ensures that the method is applicable for thin...... which mixed-mode cohesive laws are extracted....

  3. Origin of Sandwich

    Institute of Scientific and Technical Information of China (English)

    2004-01-01

    In every language there are words that have some interesting stories behind them. The word “sandwich” forexample, is very common in English. If we want to know the story behind it, we must know something about an English nobleman named Sandwich who lived in the 18th century.

  4. Making a Sandwich

    Institute of Scientific and Technical Information of China (English)

    郭富强

    2011-01-01

    Do you like eating sandwiches? Here is a recipe(做法) for a fruit sandwich.First , you should put butter(黄油)on two slices(片) of bread. Next, peel(剥开) three bananas. Now cut up(切碎) these three bananas and apple.

  5. A Numerical Investigation on the Natural Frequencies of FGM Sandwich Shells with Variable Thickness by the Local Generalized Differential Quadrature Method

    Directory of Open Access Journals (Sweden)

    Francesco Tornabene

    2017-01-01

    Full Text Available The main aim of the present paper is to solve numerically the free vibration problem of sandwich shell structures with variable thickness and made of Functionally Graded Materials (FGMs. Several Higher-order Shear Deformation Theories (HSDTs, defined by a unified formulation, are employed in the study. The FGM structures are characterized by variable mechanical properties due to the through-the-thickness variation of the volume fraction distribution of the two constituents and the arbitrary thickness profile. A four-parameter power law expression is introduced to describe the FGMs, whereas general relations are used to define the thickness variation, which can affect both the principal coordinates of the shell reference domain. A local scheme of the Generalized Differential Quadrature (GDQ method is employed as numerical tool. The natural frequencies are obtained varying the exponent of the volume fraction distributions using higher-order theories based on a unified formulation. The structural models considered are two-dimensional and require less degrees of freedom when compared to the corresponding three-dimensional finite element (FE models, which require a huge number of elements to describe the same geometries accurately. A comparison of the present results with the FE solutions is carried out for the isotropic cases only, whereas the numerical results available in the literature are used to prove the validity as well as accuracy of the current approach in dealing with FGM structures characterized by a variable thickness profile.

  6. Preparation of monoclonal antibody of anti-feline calicivirus and establishment of double-antibody sandwich enzyme-linked immunosorbent assay detecting method.

    Science.gov (United States)

    Yuan, B; Ai, C-X; Yuan, L; Gao, W; Hu, J-P; Chen, J; Ren, W-Z

    2014-09-12

    This study aimed to prepare monoclonal antibody of feline calicivirus (FCV) and identify its basic biological characteristics. Saturated ammonium sulfate precipitation, combined differential centrifugation, and cesium chloride density gradient centrifugation were used for the purification of FCV. The purified FCV was used as antigen to immunize BALB/c mice. The hybridoma lines of anti-FCV monoclonal antibodies were established using cell fusion and hybridoma screening techniques. The subtypes of the monoclonal antibody were identified. The results showed that 3 strains of hybridoma cell lines stably secreted anti-FCV monoclonal antibody; they were named as D8, E5, and H4. The D8 and E5 were IgM subtype antibodies, and H4 was IgG2b subtype antibody. The monoclonal antibody obtained shared no cross-reactivity with feline parvovirus, canine parvovirus, and canine distemper virus. According to the different recognition sites of 2 monoclonal antibodies E5 and H4 to the FCV, they were used to coat microtiter plates and prepare 2 enzyme-labeled secondary antibodies to establish double-antibody sandwich enzyme-linked immunosorbent assay detecting method.

  7. Application of "Sandwich" Teaching Method in Local Anatomy%“三明治”教学法在局部解剖学中的应用

    Institute of Scientific and Technical Information of China (English)

    李文明; 田志逢

    2016-01-01

    局部解剖学是在系统解剖学基础上,按照局部研究人体结构、形态的科学,逐层解剖人体局部。所以实践在局部解剖中尤为重要,将三明治教学法引进到局部解剖学教学中,能让学生在有限的时间内,有任务驱动的情况下,让学生真正投入到实验课堂上,达到掌握人体结构的教学目的,提高了教学效果。%The local anatomy is based on the systematic anatomy, Scientiifc research on the structure and morphology of the human body according to the local, anatomy of the human body by layer. So practice is particularly important in the local anatomy, the sandwich teaching method is introduced into the local anatomy teaching. can let the student in the limited time, there are task driven situations, so that students move up, in the experimental class, to master the teaching purpose of the structure of the human body, improve the teaching effect.

  8. ECLIA 、CMIA 、ELISA 3种方法测定血清抗CCP抗体在RA诊断中的应用%Application of three kinds of detection method ECLIA,CMIA,ELISA for detecting serum anti-CCP antibody in diagnosis of RA

    Institute of Scientific and Technical Information of China (English)

    康霞; 李欣﹟; 杨春莉; 冯平锋; 潘洁; 袁帅; 裘宇容

    2014-01-01

    目的:评价电化学发光法(ECLIA)、化学发光法(CMIA)及酶联免疫吸附测定法(ELISA)3种方法测定血清抗环瓜氨酸肽(CCP)抗体的差异及在类风湿关节炎(RA)诊断中的作用。方法采用ECLIA、CMIA及 ELISA测定45例临床确诊的RA患者、34例非RA患者及24例其他患者(包括HBsAg阳性9例、anti-HCV阳性8例、EB-IgA阳性7例)血清中抗CCP抗体水平。结果3种方法测定所有标本抗CCP抗体阳性率差异无统计学意义(P>0.05),但ELISA与ECLIA及CMIA间的一致性均较差(Kappa<0.75),ECLIA与CMIA间一致性良好(Kappa>0.75),ECLIA灵敏度优于CMIA及ELISA ,CMIA特异性优于ECLIA及ELISA。HBsAg、anti-HCV、EB-IgA阳性均可造成ELISA结果假阳性,累计阳性率达33.33%。结论在临床标本抗CCP抗体检测中,ECLIA灵敏度最高,CMIA特异性最好,ECLIA、CMIA优于ELISA ,具有重要临床应用价值。%Objective To evaluate the differences of electrochemiluminescence immunoassay (ECLIA ) ,chemiluminesence immu-noassay(CMIA) and enzyme-linked immunosorbent assay(ELISA) for detecting serum anti-CCP antibody and their role in diagno-sing rheumatoid arthritis(RA) .Methods ECLIA ,CMIA and ELISA were adopted to detect serum anti-CCP antibody in 45 patients with confirmed RA ,34 cases of non-RA and other 24 patients ,including 9 cases of positive HBsAg ,8 cases of positive anti-HCV and 7 cases of positive EB-IgA ,respectively .Results The positive rate of anti-CCP antiboy in all samples detected by ECLIA , CMIA and ELISA had no statistically significant difference (P>0 .05) .But the coincidences between ELISA with ECLIA and be-tween ELISA with CMIA were poor (Kappa 0 .75) .ECLIA showed better sensitivity than CMIA and ELISA ,while the specificity of CMIA was superior to that of ECLIA and ELISA .HB-sAg ,anti-HCV and positive EB-IgA could cause the false-positive of anti-CCP antibody detected with ELISA ,the

  9. Cornelius Elisa Bertus Bremekamp

    NARCIS (Netherlands)

    Lanjouw, J.

    1969-01-01

    Cornelis Elisa Bertus Bremekamp was born at Dordrecht on February 7th 1888. He is therefore now just past eighty, and he has been a member of the Koninklijke Nederlandse Botanische Vereniging for sixty years. He studied at the Utrecht State University and, like many of his contemporaries, was

  10. Comportamento do método quimioluminescente-ELISA em relação a resultados considerados discordantes por meio de três técnicas convencionais para diagnóstico da doença de Chagas Behavior of the chemiluminescent ELISA method in relation to results considered discordant via three conventional techniques for diagnosing Chagas disease

    Directory of Open Access Journals (Sweden)

    Cláudia Regina De Marchi

    2007-02-01

    Full Text Available Quando utilizadas, em conjunto, a hemaglutinação indireta, a imunofluorescência indireta e ELISA para diagnóstico sorológico da doença de Chagas por vezes ocorrem resultados considerados discordantes, por não haver concordância entre o que indicam essas técnicas. A disponibilidade do método quimioluminescente-ELISA permitiu executá-lo com 200 soros que examinados pelos três testes citados que motivaram a obtenção de resultados discordantes. Com o método quimioluminescente-ELISA sucederam 193 negativos e sete positivos. O emprego desse novo procedimento trouxe mais um subsídio para compreensão do assunto, mas avanço mais concreto dependerá de documentação com soros de pessoas infectadas ou não pelo Trypanosoma cruzi conforme comprovação parasitológica.When indirect hemagglutination, indirect immunofluorescence and enzyme-linked immunosorbent assay are used together for serologically diagnosing Chagas disease, results that are considered discordant sometimes occur because there is disagreement between what these tests indicate. The availability of the chemiluminescent ELISA method enabled tests on 200 serum samples that had previously produced discordant results from the three abovementioned methods. CL-ELISA revealed that 193 of these samples were negative and seven were positive. The use of this new procedure provides further support for understanding this subject, but more concrete advances will depend on documentation with blood analyses from people previously demonstrated to be unquestionably infected or uninfected with Trypanosoma cruzi.

  11. A multifunctional heat pipe sandwich panel structure

    Energy Technology Data Exchange (ETDEWEB)

    Queheillalt, Douglas T.; Wadley, Haydn N.G. [University of Virginia, Department of Materials Science and Engineering, 140 Chemistry Way, P.O. Box 400745, Charlottesville, VA 22904 (United States); Carbajal, Gerardo [University of Turabo, School of Engineering, P.O. Box 3030, Gurabo 00778 (Puerto Rico); Peterson, G.P. [University of Colorado at Boulder, 914 Broadway, Boulder, CO 80309 (United States)

    2008-01-15

    A multifunctional sandwich panel combining efficient structural load support and thermal management characteristics has been designed and experimentally assessed. The concept is based upon a truncated, square honeycomb sandwich structure. In closed cell honeycomb structures, the transport of heat from one face to the other occurs by a combination of conduction through the webs and convection/radiation within the cells. Here, much more effective heat transport is achieved by multifunctionally utilizing the core as a heat pipe sandwich panel. Its interior consists of a 6061 aluminum truncated-square honeycomb core covered with a stochastic open-cell nickel foam wick. An electroless nickel plating barrier layer inhibited the chemical reaction between the deionized water working fluid and the aluminum structure, retarding the generation of non-condensable hydrogen gas. A thermodynamic model was used to guide the design of the heat pipe sandwich panel. We describe the results of a series of experiments that validate the operational principle of the multifunctional heat pipe sandwich panel and characterize its transient response to an intense localized heat source. The systems measured thermal response to a localized heat source agrees well with that predicted by a finite difference method model used to predict the thermal response. (author)

  12. Development of an ELISA kit using monoclonal antibody to Clostridium difficile toxin A

    Institute of Scientific and Technical Information of China (English)

    Si-Wu Fu; Ya-Li Zhang; Dian-Yuan Zhou

    2004-01-01

    AIM: To establish an ELISA kit using monoclonal antibodiesagainst Clostridium difficile ( C. difficile) toxin A.METHODS: An indirect sandwich ElISA was described using the purified rabbit monospecific antiserum as capturing antibody. After the polystyrene microtitre plates with 96 fiat-bottomed wells were coated with rabbit antiserum, the wells were blocked with 100 g/L BSA in PBS-T. C. difficiletoxin A or culture filtrates were added to each well and then monoclonal antibodies IgG-horseradish peroxidase conjugate was added as detecting antibody, tetramethylbenzidine was used as substrate and A450 of the stopped reacting product was recorded in an automated plate reader. RESULTS: The tested specimens included culture filtrates of 2 strains of toxigenic C. difficile, 2 strains of non-toxigenic C. difficile, 26 strains of E. coli, 2 strains of S. dysenteriae, 1 strain of Bif infantis, 5 strains of V. cholera, 2 strains ofS. typhi, 7 strains of C. botulinum, 1 strain of toxigenic C. sordllii, and 1 strain of C. butyricum. A total of 47 strains of culture filtrates were all negative except for 2 strains of toxigenic C. difficile. The detective limitation of toxin A was 0.1 ng/mL.CONCLUSION: An ELISA kit with high specificity and excellent sensitivity for the rapid detection of C. difficile toxin A was established. It will be a useful tool for diagnostic test of C. difficile toxin A.

  13. Evaluation of a Fast and Simple Sample Preparation Method for Polybrominated Diphenyl Ether (PBDE) Flame Retardants and Dichlorodiphenyltrichloroethane (DDT) Pesticides in Fish for Analysis by ELISA Compared with GC-MS/MS.

    Science.gov (United States)

    Sapozhnikova, Yelena; Simons, Tawana; Lehotay, Steven J

    2015-05-13

    A simple, fast, and cost-effective sample preparation method, previously developed and validated for the analysis of organic contaminants in fish using low-pressure gas chromatography-tandem mass spectrometry (LPGC-MS/MS), was evaluated for the analysis of polybrominated diphenyl ethers (PBDEs) and dichlorodiphenyltrichloroethane (DDT) pesticides using enzyme-linked immunosorbent assay (ELISA). The sample preparation technique was based on the quick, easy, cheap, rugged, effective, and safe (QuEChERS) approach with filter-vial dispersive solid phase extraction (d-SPE). Incurred PBDEs and DDTs were analyzed in three types of fish with 3-10% lipid content: Pacific croaker, salmon, and National Institute of Standards and Technology (NIST) Standard Reference Material 1947 (Lake Michigan fish tissue). LPGC-MS/MS and ELISA results were in agreement: 108-111 and 65-82% accuracy ELISA versus LPGC-MS/MS results for PBDEs and DDTs, respectively. Similar detection limits were achieved for ELISA and LPGC-MS/MS. Matrix effects (MEs) were significant (e.g., -60%) for PBDE measurement in ELISA, but not a factor in the case of DDT pesticides. This study demonstrated that the sample preparation method can be adopted for semiquantitative screening analysis of fish samples by commercial kits for PBDEs and DDTs.

  14. Novel 1-D Sandwich Photonic Bandgap Structure

    Institute of Scientific and Technical Information of China (English)

    庞云波; 高葆新

    2004-01-01

    A sandwich photonic bandgap (PBG) structure is a novel PBG structure whose periodic lattice is buried in the middle of a substrate. Neither drilling nor suspending the substrate is required, and the integrity of the ground plane is maintained. This paper presents several modification techniques for sandwich PBG structure fabrication. The forbidden gap can be improved by adopting the chirping technique, applying the tapering technique, enlarging the periodic elements, adjusting the location of the periodic lattice in the substrate, and using different dielectric media H-shape elements. A finite difference time domain method is applied to analyze the structures. Deep and wide stopbands can be obtained using the modified sandwich structures. Experimental measurement results agree well with the theoretical analysis.

  15. Behaviour of Metal Foam Sandwich Panels

    DEFF Research Database (Denmark)

    2011-01-01

    Sandwich panels as used in structures comprise of a foam core enclosed by thin high strength steel faces. This paper discusses currently design formulae of local buckling behaviour of such panels using the finite element method. Multiple wave finite element models were adopted to investigate...... and examine the adequacy of currently used approach for the design of sandwich panels. The paper presents brief details of the finite element model used including geometry, load pattern and boundary conditions. The selected model gives good agreement with experimental results from Pokharel and Mahendran (2003......). The study shows that currently available design formulae are conservative for stocky sandwich plate elements while being over-conservative for high slenderness. A unified design formula of local buckling behaviour applicable to the full range of slenderness is developed....

  16. OPTIMAL DESIGN OF QUADRATIC SANDWICH PLATE

    Directory of Open Access Journals (Sweden)

    TIMAR Dr. Imre

    2016-05-01

    Full Text Available In this paper, we show the optimal design of the three-layered sandwich plates. The objective function contains the material and fabrication costs. The design constraints are the maximal stresses, the deflection of plates and damping of vibrations. The unknown is the thickness of the filling foam. By the mathematical method, we define the minima of the cost function and the optimal thickness of the filling layer of foam. The active constraint is the deflection, so we calculate of the costs of the sandwich plate with the homogeneous plate.

  17. Validation of a digital photographic method for assessment of dietary quality of school lunch sandwiches brought from home

    DEFF Research Database (Denmark)

    Sabinsky, Marianne; Toft, Ulla; Andersen, Klaus K

    2013-01-01

    from 0.89 to 0.97. The proportion of meals classified in the same or an adjacent quartile ranged from 98% (starch) to 100% (fruits, vegetables, fish, whole grain, and Meal IQ). There was no statistical difference between fish, fat, starch, whole grains, and Meal IQ using the two methods. Differences....... The lunches were photographed using a standardised DPM. From the digital images, the dietary components were estimated by a trained image analyst using weights or household measures and the dietary quality was assessed using a validated Meal Index of Dietary Quality (Meal IQ). The dietary components...

  18. DNA-based hybridization chain reaction and biotin-streptavidin signal amplification for sensitive detection of Escherichia coli O157:H7 through ELISA.

    Science.gov (United States)

    Guo, Qi; Han, Jiao-Jiao; Shan, Shan; Liu, Dao-Feng; Wu, Song-Song; Xiong, Yong-Hua; Lai, Wei-Hua

    2016-12-15

    This study reported on a novel sandwich enzyme linked immunosorbent assay (ELISA) for the sensitive determination of Escherichia coli O157:H7 (E. coli O157:H7) by using DNA-based hybridization chain reaction (HCR) and biotin-streptavidin signal amplification. The anti-E. coli O157:H7 polyclonal antibody (pAb) was immobilized in the ELISA wells. The anti-E. coli O157:H7 monoclonal antibody (mAb) and initiator strand (DNA1) were labeled on gold nanoparticle (AuNP) to form a mAb-AuNP-DNA1 complex. In the presence of the target E. coli O157:H7, the sandwiched immunocomplex, which is pAb-E. coli O157:H7-mAb-AuNP-DNA1, could be formed. Two types of biotinylated hairpin were subsequently added in the ELISA well. A nicked double-stranded DNA (dsDNA) that contained abundant biotins was formed after HCR. Detection was performed after adding horseradish peroxidase-streptavidin and substrate/chromogen solution. Under optimal conditions, E. coli O157:H7 could be detected in the range of 5×10(2) CFU/mL to 1×10(7) CFU/mL; the limit of detection was 1.08×10(2) CFU/mL in pure culture. The LOD of the novel ELISA was 185 times lower than that of traditional ELISA. The proposed method is considerably specific and can be applied in the detection of whole milk samples inoculated with E. coli O157:H7. The coefficient of variation of in pure culture and in whole milk was 0.99-5.88% and 0.76-5.38%, respectively. This method offers a promising application in the detection of low concentrations of food-borne pathogens.

  19. Behaviour of Metal Foam Sandwich Panels

    DEFF Research Database (Denmark)

    2011-01-01

    Sandwich panels as used in structures comprise of a foam core enclosed by thin high strength steel faces. This paper discusses currently design formulae of local buckling behaviour of such panels using the finite element method. Multiple wave finite element models were adopted to investigate...

  20. A phase II trial of carboplatin and docetaxel followed by radiotherapy given in a "Sandwich" method for stage III, IV, and recurrent endometrial cancer.

    Science.gov (United States)

    Geller, Melissa A; Ivy, Joseph J; Ghebre, Rahel; Downs, Levi S; Judson, Patricia L; Carson, Linda F; Jonson, Amy L; Dusenbery, Kathryn; Vogel, Rachel Isaksson; Boente, Matthew P; Argenta, Peter A

    2011-04-01

    To determine feasibility and efficacy of administering docetaxel and carboplatin chemotherapy followed by pelvic radiotherapy and then consolidation chemotherapy in patients with advanced or recurrent endometrial cancer. Patients with surgically staged III-IV (excluding IIIA from positive cytology alone) endometrial cancer or biopsy confirmed recurrent disease were eligible. Treatment consisted of 3 cycles of docetaxel (75 mg/m²) and carboplatin (AUC 6) on a q21 day schedule followed by involved field irradiation (45 Gy)± brachytherapy and three additional cycles of docetaxel and carboplatin. Kaplan-Meier (KM) methods estimated overall survival (OS) and progression free survival (PFS). Forty-two patients enrolled, 7 did not complete therapy. 95% (39/41) had primary disease. Median age=58 years (range: 21-81 years). 78% (32/41)=endometrioid histology. Stages=10 IIIA, 21 IIIC, 1 IVA, 7 IVB, (recurrent=1 IC, 1 IIA). There were 23 non-hematologic and 14 grade 3 and 16 grade 4 hematologic toxicities. Seven patients died following treatment with a median follow-up of 28 months (range: 7-70 months). KM estimates and 95% confidence intervals for OS at 1 year were 95% (82-99%), at 3 years 90% (75-96%), and at 5 years 71% (45-86%). Of the 39 with primary disease, 11 progressed or died within 5 years of study enrollment. KM estimates and 95% confidence intervals for PFS at 1 year were 87% (72-94%), at 3 years 71% (51-83%), and at 5 years 64% (42-80%). "Sandwiching" radiation between chemotherapy for advanced or recurrent endometrial cancer merits further development based on the reported PFS and OS. Copyright © 2010 Elsevier Inc. All rights reserved.

  1. Comparison of HPLC and ELISA Method in Detection of Aflatoxin M1 in Milk%HPLC与ELISA法测定牛奶中黄曲霉毒素M1的比较研究

    Institute of Scientific and Technical Information of China (English)

    胡宏灿; 韩飞; 林悦楷; 卢任杰

    2013-01-01

    The method for detecting aflatoxin Ml in milk was studied by comparison of HPLC and ELISA method. The recovery rates of HPLC were 68%-85% and the recovery rates of ELISA were 97%-109%. Detection results of HPLC and ELISA had good correlation. ELISA method had low cost, rapid detection, it could be used for preliminary screening of aflatoxin Ml. HPLC method was accurate but it had high cost of testing, it is suitable for conclusive evidence of aflatoxin Ml residue.%对HPLC法和ELISA法检测牛奶样品中黄曲霉毒素M1(AFM1)残留量进行了比较,探讨了牛奶中黄曲霉毒素M1的最佳检测方法.结果显示,对于已知加标样品(阳性),HPLC法回收率为68%~85%;ELISA法回收率为97%~109%,HPLC法和ELISA法具有良好的相关性.ELISA法检验成本较低、检测迅速,但可能出现少量的假阳性和假阴性,故可用于待测样品的初筛;HPLC法检测结果准确,但检测费用较高,适用于黄曲霉毒素M1残留的确证.

  2. Identification of Cry1Ac and Cry2Ab proteins in transgenic cotton seeds available in Gujarat (India by ELISA method

    Directory of Open Access Journals (Sweden)

    Alka Dohare

    2014-02-01

    Full Text Available Along with the increase market of the transgenic crops, the demand for testing GMOs and for certifying non-GMO foodstuffs has increased dramatically. Within the arena of expanding techniques for identification and quantification of transgenic crops, two major approaches for detecting GMOs are still applicable on large scale, namely ELISA based protein detection and PCR based gene identification. In present study, ELISA techniques was adopted to identify the specific Cry1Ac and Cry2Ab proteins in some transgenic cotton plants seed samples viz., Gujarat cotton hybrid – 6 (BG II, Gujarat cotton hybrid – 8 (BG II and Gujarat cotton hybrid – 10 (BG II from the Gujarat state of India. The study reveals the presence of both Cry1Ac and Cry2Ab proteins in the transgenic seed samples and also demonstrated that the technique of ELISA for identification of Cry1Ac and Cry2Ab proteins is quite handy and easily adoptable.

  3. Antigen capture ELISA system for henipaviruses using polyclonal antibodies obtained by DNA immunization.

    Science.gov (United States)

    Kaku, Yoshihiro; Noguchi, Akira; Marsh, Glenn A; Barr, Jennifer A; Okutani, Akiko; Hotta, Kozue; Bazartseren, Boldbaatar; Broder, Christopher C; Yamada, Akio; Inoue, Satoshi; Wang, Lin-Fa

    2012-08-01

    A novel antigen-capture sandwich ELISA system targeting the glycoproteins of the henipaviruses Nipah virus (NiV) and Hendra virus (HeV) was developed. Utilizing purified polyclonal antibodies derived from NiV glycoprotein-encoding DNA-immunized rabbits, we established a system that can detect the native antigenic structures of the henipavirus surface glycoproteins using simplified and inexpensive methods. The lowest detection limit against live viruses was achieved for NiV Bangladesh strain, 2.5 × 10(4) TCID(50). Considering the recent emergence of genetic variants of henipaviruses and the resultant problems that arise for PCR-based detection, this system could serve as an alternative rapid diagnostic and detection assay.

  4. Detecting Antibody of Tubereulosis in Cow Milk by the Method of BA-ELISA%应用BA-ELISA检测奶牛结核病乳汁抗体的研究

    Institute of Scientific and Technical Information of China (English)

    杨玉英; 李士泽; 倪宏波

    2001-01-01

    本试验建立了BA-ELISA检测牛乳中结核抗体的方法。以皮试阳性奶牛的乳样12头份,做BA-ELISA检测,结果符合率达83.33%(10/12)。结果表明该法检测牛乳结核抗体重复性好、简洁快速、敏感性高,不失为一种适用于临床的实用检测方法。%The study were conducted by establishing the method of detecting antibody of tubereulosis in cow milk by BA-ELISA. The results showed as follows: By using the way of BA-ELISA to detect 12 samples of milk by PPD positively detected, the rate of positive reaction was 83.33%.it was higher than that of routine ELISA test. The results suggested that this method has a good repetition, the operation is simple and convenient to detect antibody of tuberulosis in cow milk by using the method of BA-ELISA. So it is worth applying in practice.

  5. An ELISA for detection of antibodies against influenza A nucleoprotein in humans and various animal species.

    NARCIS (Netherlands)

    G.F. de Boer; W. Back; A.D.M.E. Osterhaus (Albert)

    1990-01-01

    textabstractA double antibody sandwich blocking ELISA, using a monoclonal antibody (MAb) against influenza A nucleoprotein (NP) was developed to detect antibodies against influenza. Collections of serum samples were obtained from human and various animal species. All influenza A subtypes induced ant

  6. Chicken IgY-based coproantigen capture ELISA for diagnosis of human opisthorchiasis.

    Science.gov (United States)

    Teimoori, Salma; Arimatsu, Yuji; Laha, Thewarach; Kaewkes, Sasithorn; Sereerak, Piya; Sripa, Manop; Tangkawattana, Sirikachorn; Brindley, Paul J; Sripa, Banchob

    2017-08-01

    Diagnosis of Opisthorchis viverrini infection by conventional stool examination is increasingly difficult due to the low intensity of the infection after several rounds of control programmes in endemic regions as well as coinfections with intestinal flukes. Therefore sensitive and specific diagnostic test is needed. In this study, a coproantigen sandwich ELISA using recombinant O. viverrini cathepsin F (rOv-CF) was developed. This sandwich ELISA employing chicken IgY raised against rOv-CF in combination with rabbit IgG antibody to the somatic O. viverrini antigens showed a lower detection limit (LLD) of 70ng native O. viverrini somatic antigens by spiking the parasite antigens into control feces. When applied to the diagnosis, the IgY-based sandwich ELISA exhibited sensitivity and specificity of 93.3% and 76.7%, respectively, in an investigation of 90 human cases positive or negative for opisthorchiasis. The positive predictive value (PPV) and negative predictive value (NPV) for this coproantigen detection were 66.7% and 95.2%, respectively. This IgY-based sandwich ELISA using parasite cathepsin F detection shows a promising immunodiagnostic alternative for human opisthorchiasis in endemic regions. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  7. Development of monoclonal antibodies and quantitative ELISAs targeting insulin-degrading enzyme

    Directory of Open Access Journals (Sweden)

    Dickson Dennis W

    2009-10-01

    Full Text Available Abstract Background Insulin-degrading enzyme (IDE is a widely studied zinc-metalloprotease implicated in the pathogenesis of type 2 diabetes mellitus, Alzheimer disease (AD and varicella zoster virus infection. Despite more than six decades of research on IDE, progress has been hampered by the lack of well-characterized reagents targeting this biomedically important protease. To address this important need, we generated and characterized new mouse monoclonal antibodies (mAbs targeting natively folded human and rodent IDE. Results Eight monoclonal hybridoma cell lines were derived in house from mice immunized with full-length, natively folded, recombinant human IDE. The mAbs derived from these lines were shown to detect IDE selectively and sensitively by a wide range of methods. Two mAbs in particular—designated 6A1 and 6H9—proved especially selective for IDE in immunocytochemical and immunohistochemical applications. Using a variety of methods, we show that 6A1 selectively detects both human and rodent IDE, while 6H9 selectively detects human, but not rodent, IDE, with both mAbs showing essentially no cross reactivity with other proteins in these applications. Using these novel anti-IDE mAbs, we also developed sensitive and quantitative sandwich ELISAs capable of quantifying IDE levels present in human brain extracts. Conclusion We succeeded in developing novel mAbs that selectively detect rodent and/or human IDE, which we have shown to be suitable for a wide range of applications, including western blotting, immunoprecipitation, immunocytochemistry, immunohistochemistry, and quantitative sandwich ELISAs. These novel anti-IDE mAbs and the assays derived from them constitute important new tools for addressing many unresolved questions about the basic biology of IDE and its role in multiple highly prevalent human diseases.

  8. Exploiting Nanobodies in the Detection and Quantification of Human Growth Hormone via Phage-Sandwich Enzyme-Linked Immunosorbent Assay

    Directory of Open Access Journals (Sweden)

    Hossam Murad

    2017-05-01

    Full Text Available BackgroundMonitoring blood levels of human growth hormone (hGH in most children with short stature deficiencies is crucial for taking a decision of treatment with extended course of daily and expensive doses of recombinant hGH (rhGH or Somatropin®. Besides, misusing of rhGH by sportsmen is banned by the World Anti-Doping Agency and thus sensitive GH-detecting methods are highly welcome in this field. Nanobodies are the tiniest antigen-binding entity derived from camel heavy chain antibodies. They were successfully generated against numerous antigens including hormones.MethodsA fully nanobody-based sandwich ELISA method was developed in this work for direct measurement of GH in biological samples.ResultsTwo major characteristics of nanobody were exploited for this goal: the robust and stable structure of the nanobody (NbGH04 used to capture hGH from tested samples, and the great ability of tailoring, enabling the display of the anti-GH detector nanobody (NbGH07 on the tip of M13-phage. Such huge, stable, and easy-to-prepare phage-Nb was used in ELISA to provide an amplified signal. Previously, NbGH04 was retrieved on immobilized hGH by phage display from a wide “immune” cDNA library prepared from a hGH-immunized camel. Here, and in order to assure epitope heterogeneity, NbGH07 was isolated from the same library using NbGH04-captured hGH as bait. Interaction of both nanobodies with hGH was characterized and compared with different anti-GH nanobodies and antibodies. The sensitivity (~0.5 ng/ml and stability of the nanobody-base sandwich ELISA were assessed using rhGH before testing in the quantification of hGH in blood sera and cell culture supernatants.ConclusionIn regard to all advantages of nanobodies; stability, solubility, production affordability in Escherichia coli, and gene tailoring, nanobody-based phage sandwich ELISA developed here would provide a valuable method for hGH detection and quantification.

  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. Standard Test Method for Measuring the Curved Beam Strength of a Fiber-Reinforced Polymer-Matrix Composite - (View Full Text) D6416/D6416M-01(2007) Standard Test Method for Two-Dimensional Flexural Properties of Simply Supported Sandwich Composite Plates Subjected to a Distributed Load

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    2001-01-01

    Standard Test Method for Measuring the Curved Beam Strength of a Fiber-Reinforced Polymer-Matrix Composite - (View Full Text) D6416/D6416M-01(2007) Standard Test Method for Two-Dimensional Flexural Properties of Simply Supported Sandwich Composite Plates Subjected to a Distributed Load

  11. Mechanical Response of All-composite Pyramidal Lattice Truss Core Sandwich Structures

    Institute of Scientific and Technical Information of China (English)

    Ming Li; Linzhi Wu; Li Ma; Bing Wang; Zhengxi Guan

    2011-01-01

    The mechanical performance of an all-composite pyramidal lattice truss core sandwich structure was investigated both theoretically and experimentally. Sandwich structures were fabricated with a hot compression molding method using carbon fiber reinforced composite T700/3234. The out-of-plane compression and shear tests were conducted. Experimental results showed that the all-composite pyramidal lattice truss core sandwich structures were more weight efficient than other metallic lattice truss core sandwich structures. Failure modes revealed that node rupture dominated the mechanical behavior of sandwich structures.

  12. Exploiting Nanobodies in the Detection and Quantification of Human Growth Hormone via Phage-Sandwich Enzyme-Linked Immunosorbent Assay.

    Science.gov (United States)

    Murad, Hossam; Assaad, Jana Mir; Al-Shemali, Rasha; Abbady, Abdul Qader

    2017-01-01

    Monitoring blood levels of human growth hormone (hGH) in most children with short stature deficiencies is crucial for taking a decision of treatment with extended course of daily and expensive doses of recombinant hGH (rhGH or Somatropin(®)). Besides, misusing of rhGH by sportsmen is banned by the World Anti-Doping Agency and thus sensitive GH-detecting methods are highly welcome in this field. Nanobodies are the tiniest antigen-binding entity derived from camel heavy chain antibodies. They were successfully generated against numerous antigens including hormones. A fully nanobody-based sandwich ELISA method was developed in this work for direct measurement of GH in biological samples. Two major characteristics of nanobody were exploited for this goal: the robust and stable structure of the nanobody (NbGH04) used to capture hGH from tested samples, and the great ability of tailoring, enabling the display of the anti-GH detector nanobody (NbGH07) on the tip of M13-phage. Such huge, stable, and easy-to-prepare phage-Nb was used in ELISA to provide an amplified signal. Previously, NbGH04 was retrieved on immobilized hGH by phage display from a wide "immune" cDNA library prepared from a hGH-immunized camel. Here, and in order to assure epitope heterogeneity, NbGH07 was isolated from the same library using NbGH04-captured hGH as bait. Interaction of both nanobodies with hGH was characterized and compared with different anti-GH nanobodies and antibodies. The sensitivity (~0.5 ng/ml) and stability of the nanobody-base sandwich ELISA were assessed using rhGH before testing in the quantification of hGH in blood sera and cell culture supernatants. In regard to all advantages of nanobodies; stability, solubility, production affordability in Escherichia coli, and gene tailoring, nanobody-based phage sandwich ELISA developed here would provide a valuable method for hGH detection and quantification.

  13. Observability and Controllability Analysis for Sandwich Systems with Backlash

    Directory of Open Access Journals (Sweden)

    Luo Na

    2015-12-01

    Full Text Available In this paper, an approach to analyze the observability and controllability of sandwich systems with backlash is proposed. In this method, a non-smooth state-space function is used to describe the sandwich systems with backlash which are also non-smooth non-linear systems. Then, a linearization method based on non-smooth optimization is proposed to derive a linearized state-space function to approximate the non-smooth sandwich systems within a bounded region around the equilibrium point that we are interested in. Afterwards, both observability and controllability matrices are constructed and the methods to analyze the observability as well as controllability of sandwich system with backlash are derived. Finally, numerical examples are presented to validate the proposed method.

  14. Comparative Study of Three Different ELISA to Measure the Antibodies Against Infectious Bronchitis Virus in Vaccinated and Unvaccinated Broilers Estudo Comparativo de Três Diferentes Modalidades de Elisa para Medir os Anticorpos Contra o Vírus da Bronquite Infecciosa em Frangos Vacinados e Não Vacinados

    OpenAIRE

    Cardoso TC; C. Oliveira; SEL Da-Silva; HL Ferreira; Pinto AA

    2001-01-01

    Broilers were spray-vaccinated (n=150) with H120 serotype at one-day-old, challenged after 28 days with M41 IBV serotype and after bled at day 28, 34 and 46 after challenged. The respective sera were tested by the indirect ELISA (I-ELISA), sandwich ELISA (S-ELISA), liquid phase blocking ELISA (LPB-ELISA) and the standard serum neutralization test (SNT). For this purpose, a total of 300 sera samples, 150 from non vaccinated and 150 from vaccinated broilers were titrated by all the serological ...

  15. Wave propagation in sandwich panels with a poroelastic core.

    Science.gov (United States)

    Liu, Hao; Finnveden, Svante; Barbagallo, Mathias; Arteaga, Ines Lopez

    2014-05-01

    Wave propagation in sandwich panels with a poroelastic core, which is modeled by Biot's theory, is investigated using the waveguide finite element method. A waveguide poroelastic element is developed based on a displacement-pressure weak form. The dispersion curves of the sandwich panel are first identified as propagating or evanescent waves by varying the damping in the panel, and wave characteristics are analyzed by examining their motions. The energy distributions are calculated to identify the dominant motions. Simplified analytical models are also devised to show the main physics of the corresponding waves. This wave propagation analysis provides insight into the vibro-acoustic behavior of sandwich panels lined with elastic porous materials.

  16. Measuring Cohesive Laws for Interfaces in Sandwich Structures

    DEFF Research Database (Denmark)

    Lundsgaard-Larsen, Christian; Sørensen, Bent F.; Berggreen, Carl Christian

    2006-01-01

    Extraction of cohesive laws are conducted for interfaces in sandwich structures. Separation between face and core are driven by pure bending moments applied to double cantilever beam (DCB) specimens. By varying the ratio between moments applied to the beams the test is conducted for different mode...... mixities. The sandwich specimens consists of glass fiber faces and Divinycell H200 foam core with a pre-crack between face and core made with teflon film. Arbitrary stiffening of the sandwich faces with steel bars adhered to the faces reduces rotations and ensures that the method is useable for a wide...

  17. Measuring Cohesive Laws for Interfaces in Sandwich Structures

    DEFF Research Database (Denmark)

    Lundsgaard-Larsen, Christian; Sørensen, Bent F.; Berggreen, Carl Christian

    2006-01-01

    Extraction of cohesive laws are conducted for interfaces in sandwich structures. Separation between face and core are driven by pure bending moments applied to double cantilever beam (DCB) specimens. By varying the ratio between moments applied to the beams the test is conducted for different mode...... mixities. The sandwich specimens consists of glass fiber faces and Divinycell H200 foam core with a pre-crack between face and core made with teflon film. Arbitrary stiffening of the sandwich faces with steel bars adhered to the faces reduces rotations and ensures that the method is useable for a wide...

  18. Sandwich Panel as a Structural Element of Overlap

    Directory of Open Access Journals (Sweden)

    Novikov Maxim

    2016-01-01

    Full Text Available This paper considers the issue of sandwich panels using as load-bearing structural elements. The comparison of deflections and critical failure loads were obtained by the results of the full-scale roof sandwich panels tests conducted by the company “Joris Ide” and the theoretical design, according to the calculation method described in Euronorms. Based on these results it was concluded that sandwich panels can be treated as a load-bearing structure only with more taught manufacturing requirements. Thus, the reduced spread of critical loads can be achieved.

  19. Procollagen type I N-terminal propeptide (PINP) as an indicator of type I collagen metabolism: ELISA development, reference interval, and hypovitaminosis D induced hyperparathyroidism

    DEFF Research Database (Denmark)

    Orum, O; Hansen, M; Jensen, Charlotte Harken;

    1996-01-01

    A sandwich enzyme-linked immunosorbent assay (ELISA) for quantification of the N-terminal propeptide of human procollagen type I (PINP) utilizing purified alpha 1-chain specific rabbit antibodies is described. The ELISA measured the content of the alpha 1-chain of PINP independent of the molecula...

  20. Structural Analysis of Sandwich Foam Panels

    Energy Technology Data Exchange (ETDEWEB)

    Kosny, Jan [ORNL; Huo, X. Sharon [Tennessee Technological University

    2010-04-01

    The Sandwich Panel Technologies including Structural Insulated Panels (SIPs) can be used to replace the conventional wooden-frame construction method. The main purpose of this Cooperative Research and Development Agreement (CRADA) between UT-Battelle, LLC and SGI Venture, Inc. was to design a novel high R-value type of metal sandwich panelized technology. This CRADA project report presents design concept discussion and numerical analysis results from thermal performance study of this new building envelope system. The main objective of this work was to develop a basic concept of a new generation of wall panel technologies which will have R-value over R-20 will use thermal mass to improve energy performance in cooling dominated climates and will be 100% termite resistant. The main advantages of using sandwich panels are as follows: (1) better energy saving structural panels with high and uniform overall wall R-value across the elevation that could not be achieved in traditional walls; and (2) reducing the use of raw materials or need for virgin lumber. For better utilization of these Sandwich panels, engineers need to have a thorough understanding of the actual performance of the panels and system. Detailed analysis and study on the capacities and deformation of individual panels and its assembly have to be performed to achieve that goal. The major project activity was to conduct structural analysis of the stresses, strains, load capacities, and deformations of individual sandwich components under various load cases. The analysis simulated the actual loading conditions of the regular residential building and used actual material properties of the steel facings and foam.

  1. Compressive and shear buckling analysis of metal matrix composite sandwich panels under different thermal environments

    Science.gov (United States)

    Ko, William L.; Jackson, Raymond H.

    1993-01-01

    Combined inplane compressive and shear buckling analysis was conducted on flat rectangular sandwich panels using the Raleigh-Ritz minimum energy method with a consideration of transverse shear effect of the sandwich core. The sandwich panels were fabricated with titanium honeycomb core and laminated metal matrix composite face sheets. The results show that slightly slender (along unidirectional compressive loading axis) rectangular sandwich panels have the most desirable stiffness-to-weight ratios for aerospace structural applications; the degradation of buckling strength of sandwich panels with rising temperature is faster in shear than in compression; and the fiber orientation of the face sheets for optimum combined-load buckling strength of sandwich panels is a strong function of both loading condition and panel aspect ratio. Under the same specific weight and panel aspect ratio, a sandwich panel with metal matrix composite face sheets has much higher buckling strength than one having monolithic face sheets.

  2. Enhanced sandwich immunoassay using antibody-functionalized magnetic iron-oxide nanoparticles for extraction and detection of soluble transferrin receptor on a photonic crystal biosensor.

    Science.gov (United States)

    Peterson, Ross D; Chen, Weili; Cunningham, Brian T; Andrade, Juan E

    2015-12-15

    Iron deficiency anemia (IDA) has detrimental effects on individuals and societies worldwide. A standard sandwich assay (SA) for the detection of soluble transferrin receptor (sTfR), a biomarker of IDA, on a photonic crystal (PC) biosensor was established, but it was susceptible to non-specific signals from complex matrixes. In this study, iron-oxide nanoparticles (fAb-IONs) were used as magnetic immuno-probes to bind sTfR and minimize non-specific signals, while enhancing detection on the PC biosensor. This inverse sandwich assay (IA) method completely bound sTfR with low variability (detection in sera (Liquichek™ control sera) on the PC biosensor using two certified ELISAs as reference methods. A linear dose-response curve was elicited at the fAb-IONs concentration in which the theoretical binding ratio (sTfR:fAb-IONs) was calculated to be 0.05) at 14 and 21 μg/mL, respectively. The inherent imprecision of the IA and reference ELISAs was σ(δ)=0.45 µg/mL and the mean biases for Liquichek™ 1, 2 and 3 were 0.18, 0.19 and -0.04 µg/mL, respectively. Whereas the inherent imprecision of the SA and reference ELISAs was σ(δ)=0.52 µg/mL and the biases for Liquichek™ 1, 2 and 3 were 0.66, 0.14 and -0.67 µg/mL, respectively. Thus, unlike the SA, the IA method measures sTfR with the same bias as the reference ELISAs. Combined magnetic separation and detection of nutrition biomarkers on PC biosensors represents a facile method for their accurate and reliable quantification in complex matrixes.

  3. b型流感嗜血杆菌多糖竞争ELISA检测方法的建立与应用%Establishment and application of competitive ELISA method for detertion of Haemophilus influenzae type b polysaccharide

    Institute of Scientific and Technical Information of China (English)

    刘威; 江山; 杨溢尧; 李小波; 兰芳; 崔长法

    2014-01-01

    目的 建立检测b型流感嗜血杆菌(Haemophilus influenzae type b,Hib)多聚核糖基核糖醇磷酸盐(polyribosylribitol phosphate,PRP)的竞争酶联免疫吸附法(competative enzyme-linked immunosorbent assay,cELISA).方法 以Hib PRP-酪胺(PRP-Ty)为包被抗原,待测PRP为竞争抗原,利用方阵滴定法确定抗原与血清抗PRP抗体的最适反应条件,建立cELISA法并验证其准确性和精密度,并将该法与传统检测法进行比较.结果 最适包被PRP-Ty浓度为1.30 mg/L,血清抗PRP抗体稀释度为1∶40 000.该法的检测线性范围为6.25~100.00 mg/L,决定系数(R2)为0.99,批内精密度为3.39%~6.53%,批间精密度为8.48%.该法对Hib PRP的检测结果与传统化学法一致.结论 建立的Hib多糖cELISA法的准确性和精密性良好,可特异性检测Hib PRP.%Objective To establish competitive ELISA (cELISA) method for determining polyribosylribitol phosphate (PRP) of Haemophilus influenzae type b (Hib).Methods PRP-Ty was used as coating antigen and Hib PRP sample was used as competitive antigen.Optimal reaction conditions were determined between antigens and serum antibodies to Hib PRP by criss-cross serial dilution analysis.The cELISA method was established and its accuracy and precision were validated.The cELISA method was compared with the traditional method.Results Optimal coating concentration of PRP-Ty and antiserum dilution were 1.30 mg/L and 1 ∶ 40 000,respectively.The linear range of the cELISA method was from 6.25 mg/L to 100.00 mg/L,and the coefficient of determination (R2) was 0.99.The precision of intraassay and inter-assay were 3.39%-6.53% and 8.48%,respectively.The detection results of Hib PRP with the cELISA method were consistent with those with the traditional chemical method.Conclusion The cELISA method has better accuracy and precision,and can be used for detection of Hib polysaccharide.

  4. ELISA法与TPPA法检测梅毒螺旋体抗体的相关性研究%Relativity of ELISA and TPPA methods in detection of treponema pallidum antibody

    Institute of Scientific and Technical Information of China (English)

    魏方; 张鹏

    2014-01-01

    Objective:To investigate the relativity of enzyme-linked immunosorbent assay(ELISA) and treponema pallidum antibody gelatin particle agglutination test( TPPA) in detection of syphilis. Methods:Treponema pallidum antibodies in the serum were detected by ELISA,and a total of 207 positive samples and 50 negative samples were collected;then the collected samples were confirmed by TPPA test. According to the s/co value,the patients were divided into four groups and the positive rate of each group was analyzed by TPPA method. Results:The treponema pallidum antibody in the sera of 50 patients with s/co value<1 by ELISA were all negative when detected by TPPA;the positive rates of the serum treponema pallidum antibody in patients with s/co value of 1 to 3. 99,4 to 9. 99 and≥10 by ELISA were 88. 7%,98. 6% and 100. 0%,respectively. The positive rate of treponema pallidum antibody by TPPA test increased with the s/co value by ELISA method(P <0. 01). Conclusions:ELISA is a highly sensitive and automated method in detection of syphilis, which is suitable for clinical routine screening of a large number of specimens. Test by ELISA and TPPA is recommended when s/co value is between 1 and 9. 99 in syphilis detection by ELISA to reduce the false positive possibility and provide more accurate laboratory evidence for clinical diagnosis and treatment of syphilis.%目的:探讨酶联免疫吸附试验( ELISA)与梅毒螺旋体抗体明胶颗粒凝集试验( TPPA)对梅毒螺旋体抗体检测的相关性。方法:采用ELISA法对患者血清进行梅毒螺旋体抗体检测,收集阳性血清标本207例,阴性标本50例,使用TPPA法进行确认;根据测定的s/co值将患者分为4组,分析各组TPPA的阳性率。结果:50份ELISA测定值s/co<1的样本经TPPA检测均为阴性;62份ELISA测定值s/co为1~3.99的样本经TPPA检测阳性率为88.7%;80份ELISA测定值s/co为4~9.99的样本经TPPA检测阳性率为98.6%;65份ELISA测定值s/co≥10的

  5. The asymmetric sandwich theorem

    CERN Document Server

    Simons, Stephen

    2011-01-01

    We discuss the asymmetric sandwich theorem, a generalization of the Hahn-Banach theorem. As applications, we derive various results on the existence of linear functionals that include bivariate, trivariate and quadrivariate generalizations of the Fenchel duality theorem. Most of the results are about affine functions defined on convex subsets of vector spaces, rather than linear functions defined on vector spaces. We consider both results that use a simple boundedness hypothesis (as in Rockafellar's version of the Fenchel duality theorem) and also results that use Baire's theorem (as in the Robinson-Attouch-Brezis version of the Fenchel duality theorem). This paper also contains some new results about metrizable topological vector spaces that are not necessarily locally convex.

  6. Fracture of sandwiched composites

    Energy Technology Data Exchange (ETDEWEB)

    Lin, Weh-Huei.

    1989-01-01

    Fracture of a pair of collinear cracks in various materials, such as an isotropic strip, an orthotropic strip, a bonded isotropic adhesive layer, and sandwiched orthotropic layers, is investigated. The crack surfaces are subjected to an arbitrary opening pressure p(x). The problems are formulated in terms of Fredholm integral equation of the second kind by making use the techniques of Fourier transform and finite Hilbert transform. In case of uniform opening pressure p(x)={sigma}, exact expressions for the stress intensity factors and the shape of deformed crack are obtained. Numerical calculations are carried out to study the effects of various boundary geometries and material properties on the fracture of the chosen materials.

  7. Fracture Characterization of Sandwich Face/Core Interfaces

    DEFF Research Database (Denmark)

    Manca, Marcello

    such result it is important to devise new experimental and analytical techniques to establish the multi-mode fracture characteristics of sandwich plate structures and accordingly develop methods to inhibit defect propagation. This thesis deals with characterization of fracture between face and core...... samples. A number of sandwich materials were tested (GFRP/foam cores and CFRP/Nomex) bothin static and fatigue. A linear elastic fracture mechanics model was used to determine the analyticalexpression of compliance which allowed to calculate automatically the crack length. In combination, a finite element...... types of sandwich samples atdifferent mode mixities and ΔG levels, and results in agreement with previous studies were obtained suggesting that the developed testing method is a reliable tool for the study of face/core interface debonded sandwich structures....

  8. An overview on ELISA techniques for FMD

    Directory of Open Access Journals (Sweden)

    Ding Yao-zhong

    2011-09-01

    Full Text Available Abstract Background FMD is one of the major causes of economic loss of cloven-hoofed animals in the world today. The assessment of dominant genotype/lineage and prevalent trends and confirmation the presence of infection or vaccination not only provides scientific basis and first-hand information for appropriate control measure but also for disease eradication and regaining FMD free status following an outbreak. Although different biological and serological approaches are still applied to study this disease, ELISA test based on the distinct format, antigen type and specific antibody reinforce its predominance in different research areas of FMD, and this may replace the traditional methods in the near future. This review gives comprehensive insight on ELISA currently available for typing, antigenic analysis, vaccination status differentiation and surveillance vaccine purity and content at all stages of manufacture in FMDV. Besides, some viewpoint about the recent advances and trends of ELISA reagent for FMD are described here. Methods More than 100 studies regarding ELISA method available for FMD diagnosis, antigenic analysis and monitor were thoroughly reviewed. We investigated previous sagacious results of these tests on their sensitivity, specificity. Results We found that in all ELISA formats for FMD, antibody-trapping and competitive ELISAs have high specificity and RT-PCR (oligoprobing ELISA has extra sensitivity. A panel of monoclonal antibodies to different sites or monoclonal antibody in combination of antiserum is the most suitable combination of antibodies in ELISA for FMD. Even though from its beginning, 3ABC is proven to be best performance in many studies, no single NSP can differentiate infected from vaccinated animals with complete confidence. Meanwhile, recombinant antigens and peptide derived from FMDV NPs, and NSPs have been developed for use as an alternative to the inactivated virus antigen for security. Conclusions There is

  9. Development of an Aquaporin-4 Orthogonal Array of Particle-Based ELISA for Neuromyelitis Optica Autoantibodies Detection.

    Directory of Open Access Journals (Sweden)

    Francesco Pisani

    Full Text Available Serological markers of Nuromyelitis Optica (NMO, an autoimmune disorder of the central nervous system, are autoantibodies targeting the astrocytic water channel aquaporin-4 (AQP4. We have previously demonstrated that the main epitopes for these autoantibodies (AQP4-IgG are generated by the supramolecular arrangement of AQP4 tetramers into an Orthogonal Array of Particles (OAPs. Many tests have been developed to detect AQP4-IgG in patient sera but several procedural issues affect OAP assembly and consequently test sensitivity. To date, the protein based ELISA test shows the lowest sensitivity while representing a valid alternative to the more sensitive cell based assay (CBA, which, however, shows economic, technical and interpretation problems. Here we have developed a high perfomance ELISA in which native OAPs are used as the molecular target. To this aim a native size exclusion chromatography method has been developed to isolate integral, highly pure and AQP4-IgG-recognized OAPs from rat brain. These OAPs were immobilized and oriented on a plastic plate by a sandwich approach and 139 human sera were tested, including 67 sera from NMO patients. The OAP-ELISA showed a 99% specificity and a higher sensitivity (91% compared to the CBA test. A comparative analysis revealed an end-point titer three orders of magnitude higher than the commercial ELISA and six times higher than our in-house CBA test. We show that CNS-extracted OAPs are crucial elements in order to perform an efficient AQP4-IgG test and the OAP-ELISA developed represents a valid alternative to the CBA currently used.

  10. Development of an Aquaporin-4 Orthogonal Array of Particle-Based ELISA for Neuromyelitis Optica Autoantibodies Detection.

    Science.gov (United States)

    Pisani, Francesco; Settanni, Paolo; Rosito, Stefania; Mola, Maria Grazia; Iorio, Raffaele; Tortorella, Carla; Ruggieri, Maddalena; Trojano, Maria; Svelto, Maria; Frigeri, Antonio; Nicchia, Grazia Paola

    2015-01-01

    Serological markers of Nuromyelitis Optica (NMO), an autoimmune disorder of the central nervous system, are autoantibodies targeting the astrocytic water channel aquaporin-4 (AQP4). We have previously demonstrated that the main epitopes for these autoantibodies (AQP4-IgG) are generated by the supramolecular arrangement of AQP4 tetramers into an Orthogonal Array of Particles (OAPs). Many tests have been developed to detect AQP4-IgG in patient sera but several procedural issues affect OAP assembly and consequently test sensitivity. To date, the protein based ELISA test shows the lowest sensitivity while representing a valid alternative to the more sensitive cell based assay (CBA), which, however, shows economic, technical and interpretation problems. Here we have developed a high perfomance ELISA in which native OAPs are used as the molecular target. To this aim a native size exclusion chromatography method has been developed to isolate integral, highly pure and AQP4-IgG-recognized OAPs from rat brain. These OAPs were immobilized and oriented on a plastic plate by a sandwich approach and 139 human sera were tested, including 67 sera from NMO patients. The OAP-ELISA showed a 99% specificity and a higher sensitivity (91%) compared to the CBA test. A comparative analysis revealed an end-point titer three orders of magnitude higher than the commercial ELISA and six times higher than our in-house CBA test. We show that CNS-extracted OAPs are crucial elements in order to perform an efficient AQP4-IgG test and the OAP-ELISA developed represents a valid alternative to the CBA currently used.

  11. 复合材料泡沫夹层结构的缺陷评定方法研究%Research on the Evaluation Method of Defects for Composite Foam Sandwiches

    Institute of Scientific and Technical Information of China (English)

    唐桂云; 王云飞; 于柏峰

    2011-01-01

    本文以厚壁碳纤维复合材料为面板,硬质聚氨酯泡沫为芯材制造复合材料泡沫夹层结构,模拟实际生产过程中容易出现的面板与芯材之间界面的脱粘和界面胶层过厚的现象,采用人工制造试块的方法,研究了超声波探伤对夹层复合材料缺陷的评定方法,解决了实际检测过程中的疑问,为夹层复合材料结构产品的质量检验提供依据。得出了粘接良好区胶层过厚不会被判定为脱粘的结论。%The potential debond and the thicker interfacial bonding layer between the panel and the core during the practical processing were simulated for the foam sandwich structures with thick carbon fiber panels and rigid polyurethane foam core. Using the man - made defect method, the evaluating method of sandwich composite defects was researched with ultrasonic testing, which solved the problems in the practical testing process and provided the evidence for the quality tes- ting of sandwich composite structures. The conclusions were got that the thicker bonding layers in the better adhesive area will not be considered as the debondinz.

  12. Gold nanoparticle-based enzyme-linked antibody-aptamer sandwich assay for detection of Salmonella Typhimurium.

    Science.gov (United States)

    Wu, Wenhe; Li, Jun; Pan, Dun; Li, Jiang; Song, Shiping; Rong, Mingge; Li, Zixi; Gao, Jimin; Lu, Jianxin

    2014-10-01

    Enzyme-linked immunosorbent assay (ELISA) provides a convenient means for the detection of Salmonella enterica serovar Typhimurium (STM), which is important for rapid diagnosis of foodborne pathogens. However, conventional ELISA is limited by antibody-antigen immunoreactions and suffers from poor sensitivity and tedious sample pretreatment. Therefore, development of novel ELISA remains challenging. Herein, we designed a comprehensive strategy for rapid, sensitive, and quantitative detection of STM with high specificity by gold nanoparticle-based enzyme-linked antibody-aptamer sandwich (nano-ELAAS) method. STM was captured and preconcentrated from samples with aptamer-modified magnetic particles, followed by binding with detector antibodies. Then nanoprobes carrying a large amount of reporter antibodies and horseradish peroxidase molecules were used for colorimetric signal amplification. Under the optimized reaction conditions, the nano-ELAAS assay had a quantitative detection range from 1 × 10(3) to 1 × 10(8) CFU mL(-1), a limit of detection of 1 × 10(3) CFU mL(-1), and a selectivity of >10-fold for STM in samples containing other bacteria at higher concentration with an assay time less than 3 h. In addition, the developed nanoprobes were improved in terms of detection range and/or sensitivity when compared with two commercial enzyme-labeled antibody signal reporters. Finally, the nano-ELAAS method was demonstrated to work well in milk samples, a common source of STM contamination.

  13. Forced vibration of a shear thickening fluid sandwich beam

    Science.gov (United States)

    Wei, Minghai; Hu, Gang; Jin, Lu; Lin, Kun; Zou, Dujian

    2016-05-01

    The forced vibration of a sandwich beam integrating a shear thickening fluid (STF) core and with conductive skins subjected to a periodic excitation was investigated theoretically in this study. The rheological properties of the STF material including viscosity, plasticity, and elasticity may be changed under the periodic vibration, and hence they were considered. The governing equation of motion was derived based on the complex stiffness method and some key parameters were derived based on the Timoshenko beam theory. Effects of the excitation frequency, the excitation amplitude, the excitation location, and the skin/core thickness ratio on the nature frequency of the sandwich beam were investigated. It was found that the STF core has a significant effect on the dynamic property of the sandwich beam. Based on the findings, integrating the STF core in a sandwich beam can reduce the vibration of the beam.

  14. Development of Aircraft Sandwich Parts

    Directory of Open Access Journals (Sweden)

    J. Křena

    2000-01-01

    Full Text Available The presented paper shows the design and development process of sandwich parts. A spoiler plate and a main landing gear door are developed. Sandwich parts are made of C/E composite facings and a foam core. FE models have been used for optimization of structures. Emphasis has been placed on deformations of parts under a few load cases. Experimental tests have been used for a verification of structure parts loaded by concentrated forces.

  15. Establishment of a double antibody sandwich ELISA method to detect Avian Influenza Virus (AIV)%检测禽流感病毒抗原的双抗体夹心ELISA方法的建立

    Institute of Scientific and Technical Information of China (English)

    张立怀; 张国中; 霍蕾; 侯艳梅

    2009-01-01

    禽流行性感冒(avian influenza,AI)简称禽流感,是由正粘病毒科流感病毒属A型流感病毒引起的禽类传染病。高致病性禽流感(highly pathogenic avian influenza,HPAI)被世界动物卫生组织(world organization for animal health,OIE)列为A类传染病。我国也把禽流感列为一类动物疫病。禽流感病毒(avian influenza virus,AIV)可感染几乎所有野生禽及家禽,

  16. Investigation on Stake Welding Method of T-joints for Metal Sandwich Panel%金属夹芯结构制造中的T型接头Stake焊接方法研究

    Institute of Scientific and Technical Information of China (English)

    谷侃锋; 魏强; 赵明扬

    2012-01-01

    The Stake welding method of T-joints is a critical technology for making metal sandwich panels. In order to demonstrate the feasibility of making metal sandwich panels with welding method, three welding methods, including laser welding, gas tungsten arc (TIG) welding, and laser-TIG hybrid welding with cold welding wire, were investigated experimentally on Stake welding method of titanium alloy thin sheet T-joints. The weld forming properties of the T-joints and some affecting factors were analyzed under this three welding methods. The results show that the laser-TIG hybrid welding with cold welding wire method is an ideal technical for the Stake welding method of T-joints for making metal sandwich panels, which has more advantages than the laser welding method and the gas tungsten arc(TIG) welding method in the microstructure of welded joint, efficiency of welding, weld shaping and adaptability of weld gap.%T型接头的Stake焊接方法是实现金属夹芯板焊接制造的关键技术之一.为了论证焊接制造金属夹芯板的可行性,本文通过实验对比研究了在单一激光、TIG电弧和激光-TIG电弧复合等三种焊接工艺下,钛合金薄板T型接头Stake焊接焊缝成形特点及影响因素.结果表明,激光-TIG电弧焊是实现钛合金薄板T型接头Stake焊接的理想工艺,在焊缝组织、焊接效率、焊缝成形、间隙适应性等方面比单一激光或TIG焊接工艺有明显的优势,可以应用于金属夹芯板的焊接制造.

  17. Detection of hazelnuts and almonds using commercial ELISA test kits.

    Science.gov (United States)

    Garber, Eric A E; Perry, Jesse

    2010-03-01

    Three commercial sandwich enzyme-linked immunosorbent assay (ELISA) test kits for the detection of hazelnuts and almonds were evaluated. Limits of detection and dynamic ranges were determined for hazelnuts and almonds spiked into cooked oatmeal, dipping chocolate, and muffins (baked). The limit of detection values varied from 1 to 38 μg/g, depending on the food matrix and ELISA test kit. Percent recoveries based on the standards supplied with the test kits varied from 10% to 170%. It is impossible to ascertain whether the percent recoveries reflect the performance of the ELISAs or differences between the protein content of the nuts used to spike the samples and the test kit standards. Unfortunately, reference materials do not exist that can be used to compare the results from different test kits and standardize the test kit standards. Also, insufficient knowledge regarding the epitope specificity of the antibodies used in the ELISAs further hinders interpretation of the results generated by the different test kits.

  18. Optimization of composite sandwich cover panels subjected to compressive loadings

    Science.gov (United States)

    Cruz, Juan R.

    1991-01-01

    An analysis and design method is presented for the design of composite sandwich cover panels that include the transverse shear effects and damage tolerance considerations. This method is incorporated into a sandwich optimization computer program entitled SANDOP. As a demonstration of its capabilities, SANDOP is used in the present study to design optimized composite sandwich cover panels for for transport aircraft wing applications. The results of this design study indicate that optimized composite sandwich cover panels have approximately the same structural efficiency as stiffened composite cover panels designed to satisfy individual constraints. The results also indicate that inplane stiffness requirements have a large effect on the weight of these composite sandwich cover panels at higher load levels. Increasing the maximum allowable strain and the upper percentage limit of the 0 degree and +/- 45 degree plies can yield significant weight savings. The results show that the structural efficiency of these optimized composite sandwich cover panels is relatively insensitive to changes in core density. Thus, core density should be chosen by criteria other than minimum weight (e.g., damage tolerance, ease of manufacture, etc.).

  19. [A simple ELISA method for the detection of HBsAg: Organon Teknika HBsAg Uniform II screening and confirmation test. Comparative study using the HBsAg Hapanostika method. A multicenter study].

    Science.gov (United States)

    Pár, A; Mihály, I; Kömives, K

    1994-09-25

    An one-step enzyme-linked immunoabsorbent method, named as HBsAg Uniform II has been described for the detection of serum HBsAg, and a comparison was made with a widely used ELISA technique HBsAg Hepanostika test, to evaluate sensitivity, specificity and reproducibility of the method. A total of 531 serum samples from patients with liver disease and with renal failure, as well as 1065 samples from blood donors have been investigated. While the sensitivity of Uniform II vs. Hepanostika was 99.5% vs. 72.7%, the specificity was 99.2% of both methods. The positive predictive values did not differ (99.5% vs. 99.2%), however, the negative predictive values were 99.2% vs. 71.7%, respectively, in favour of Uniform II test. The Uniform II confirmatory test confirmed the positive HBsAg results in 94%, this rate was 74% using Hepanostika system. The new method proved to be a simple, quick, reliable test, which can be useful as a valuable tool in both the clinical diagnosis and blood donor screening.

  20. Elisa Biagini (Florencia, 1970

    Directory of Open Access Journals (Sweden)

    Berta González Saavedra

    2017-01-01

    Full Text Available Elisa Biagini vive en Italia tras haber estudiado y enseñado en los Estados Unidos durante varios años. Sus poesías han sido publicadas en varias revistas y antologías italianas y americanas, entre otras. Algunas de las más recientes son Nuovissima poesia italiana (Mondadori, 2004 y Parola plurale (Sosselam 2005. Ha publicado siete colecciones poéticas, algunas bilingües, entre las que figuran  L'Ospite (Einaudi, 2004, Fiato. Parole per musica (Edizionidif, 2006, Nel bosco (Einaudi, 2007, The guest in the wood (Chelsea editions, 2013 – 2014 Best Translated Book Award, y la reciente Da una crepa (Einaudi, 2014. Sus poesías han sido traducidas al inglés, español, francés, portugués, japonés, croata, eslovaco, alemán, albanés, ruso, árabe y chino. Ha participado en importantes festivales italianos e internacionales (entre otros, en Italia “Festival della Letteratura” de Mantua, “Festival Poesia” de Parma, “RomaPoesia” de Roma, y en el extranjero “Stanza- Scotland's International Poetry Festival” en St. Andrews, Escocia, “Dubai International Poetry Festival” en Emiratos Árabes Unidos, “PoesieFestivalBerlin” en Berlín, “International Writers Workshop” en Hong Kong, “Struga Poetry Festival” en Struga, Macedonia, “Poetry Parnassus” en Londres, “Printemps des poètes” en Luxemburgo, “Queensland Poetry Festival” en Brisbane, Australia, “Festival Internacional de Poesía de Granada” en Nicaragua, “Xu Zhimo Poetry and Art Festival” en el King's College de Cambridge. Asimismo es traductora de poesía americana y, además de editar algunas colecciones de poetisas americanas contemporáneas, se ha encargado de la edición del volumen Nuovi poeti americani (Einaudi, 2006. Infine, insegna Scrittura Creativa (poesia, Travel Writing e Storia dell'Arte in Italia e all’estero, además de colaborar con artistas visules, coreógrafos y músicos. Entre otras actividades, es artista visual. www.elisabiagini.it

  1. 金标法和ELISA法检测乙型肝炎病毒血清标志物的比较%Comparative Study on Colloidal Gold-labeled Method and ELISA of Testing Five Serological Markers of HBV

    Institute of Scientific and Technical Information of China (English)

    丁邦胜; 潘健

    2012-01-01

    目的 探讨金标法检测乙型肝炎病毒(HBV)HBsAg、HBsAb、HBeAg、HBeAb、HBcAb五项血清标志物的敏感性、特异性.方法 采用ELISA法和金标法对256例血清样本同时进行HBV五项血清标志物的检测,并且对结果进行对比分析.结果 256例样本中:ELISA法“全阴性”(HBsAg,HBsAb,HBeAg,HBeAb,HBcAb均阴性)结果模式,金标法结果相同;ELISA法HBsAg、HBeAb、HBcAb阳性,HBsAb、HBeAg阴性(简称模式1);HBsAg、HBeAg、HBcAb阳性,HBeAb、HBsAb阴性(简称模式2)结果模式,金标法检测样本的结果模式发生改变.256例样本两法各单项指标检测结果经x2检验:HBsAb和HBeAg两项结果的差异无统计学意义(P>0.05);HB-sAg,HBeAb,HBcAb三项结果的差异有统计学意义(P<0.05),少数ELISA法阳性或OD为临界值的样本金标法检测结果为阴性;无ELISA法阴性而金标法为阳性的标本.结论 两种方法对HBV五项血清标志物测定的特异性相近,但是金标法敏感性不如ELISA法.%Objective To investigate the sensitivity and specificity of testing HBsAg. HBsAb, HBeAg, HBeAb and HBcAb with colloidal gold-labeled method. Methods 256 serological cases were tested with both colloidal gold-labeled method and ELISA. The results of two methods were analyzed. Results All five negative results samples by ELISA were the same with colloidal gold-labeled method.but for the results HBsAg. HBeAb, HBcAb are positive)HBsAb,HBeAg are negative and HBsAg.HBeAg. HBcAb are positive; HBeAb.HBsAb are negative by ELISA. some were changed by colloidal gold-labeled method. The chi-aquare test(x2) showed that the result differences of HBsAb and HBeAg were not statistically significant with two methods (P>0.05 ). But X2 test showed the differences were significant on HBsAg,HBeAb and HBcAb with two methods. The weak positive results by ELISA always were negative by colloidal gold-labeled method. but no negative results by ELISA were found positive tested by colloidal gold

  2. Development of monoclonal antibodies and quantitative sandwich enzyme linked immunosorbent assay for the characteristic sialoglycoprotein of edible bird's nest.

    Science.gov (United States)

    Zhang, Shiwei; Lai, Xintian; Liu, Xiaoqing; Li, Yun; Li, Bifang; Huang, Xiuli; Zhang, Qinlei; Chen, Wei; Lin, Lin; Yang, Guowu

    2013-01-01

    The article presents a sandwich enzyme linked immunosorbent assay (ELISA) for identification of edible bird's nest. The characteristic sialoglycoproteins were found by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and purified by liquid-phase isoelectric focusing (LIEF). According to the analysis, the molecular weight was 106-128 kDa and the isoelectric point was ≤pH 3.0. Two anti-characteristic sialoglycoprotein monoclonal antibodies were produced. The monoclonal antibodies were examined by western-blot assay. One of the monoclonal antibody was used as coating and the other as the enzyme-labeled antibody after being coupled to horseradish peroxidase (HRP). Based on the optimized ELISA condition, the method was established with IC(50) of 1.5 ng/mL, and low cross-reactivity with various fake materials (ELISA provided a suitable means for screening of a large number of samples. The coefficients of variation were between 2.9% and 5.8%.

  3. Influence of film thickness on structural and optical properties of ZnS thin films obtained by SILAR method and analysis of Zn/ZnS/n-GaAs/In sandwich structure

    Energy Technology Data Exchange (ETDEWEB)

    Oezakin, Oguzhan; Guezeldir, Betuel; Saglam, Mustafa [Department of Physics, Science Faculty, Atatuerk University, Erzurum (Turkey); Yildirim, M. Ali [Department of Physics, Science and Art Faculty, Erzincan University, Erzincan (Turkey); Ates, Aytunc [Department of Material Engineering, Faculty of Engineering and Natural Sciences, Yildirim Beyazit University, Ankara (Turkey)

    2012-04-15

    ZnS thin films were deposited on glass substrates using SILAR method at room temperature and ambient pressure. The relationship between refractive index and energy bandgap was investigated. The film thickness effect on the structural, morphological and optical properties of ZnS thin films was investigated. The crystalline and surface properties of the films improved with increasing film thickness. The energy bandgap values changed from 3.87 to 3.58 eV with increasing film thickness. The refractive index (n), high frequency dielectric constant ({epsilon}{sub {infinity}}) values were calculated by using the energy bandgap values as a function of the film thickness. Also, ZnS thin film was deposited directly on n-GaAs substrate for obtaining the Zn/ZnS/n-GaAs/In sandwich structure at room temperature. The sandwich structure demonstrated clearly rectifying behaviour by the current-voltage (I-V) curves at room temperature. From I-V characteristics n and {phi}{sub b} values were calculated as 1.894 and 0.632 eV at room temperature, respectively. (Copyright copyright 2012 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

  4. Comparative evaluation of conventional methods and elisa based IgG antibodies detection for diagnosis of helicobacter pylori infection in cases of dyspepsia

    Directory of Open Access Journals (Sweden)

    Arora U

    2003-01-01

    Full Text Available Seventy five gastric biopsy specimens and 75 serum samples of same patients complaining of dyspepsia were collected. Biopsy specimens were processed for rapid urease test, gram staining and culture. Serum samples were used for detecting IgG antibodies against 128kDa external protein (Cog A of H.pylori using a commercially available ELISA kit. Rapid urease test was positive in 54 (72%, culture in 21 (28% and gram staining in 15 (20%. Significant IgG levels were detected in 57 (76% cases. It was therefore concluded that for diagnosis of H.pylori infection in cases of dyspepsia, determination of IgG levels can act as an important screening procedure.

  5. A Sandwich HIV p24 Amperometric Immunosensor Based on a Direct Gold Electroplating-Modified Electrode

    Directory of Open Access Journals (Sweden)

    Ning Gan

    2012-05-01

    Full Text Available Acquired immune deficiency syndrome (AIDS is a severe communicable immune deficiency disease caused by the human immune deficiency virus (HIV. The analysis laboratory diagnosis of HIV infection is a crucial aspect of controlling AIDS. The p24 antigen, the HIV-1 capsid protein, is of considerable diagnostic interest because it is detectable several days earlier than host-generated HIV antibodies following HIV exposure. We present herein a new sandwich HIV p24 immunosensor based on directly electroplating an electrode surface with gold nanoparticles using chronoamperometry, which greatly increased the conductivity and reversibility of the electrode. Under optimum conditions, the electrochemical signal showed a linear relationship with the concentration of p24, ranging from 0.01 ng/mL to 100 ng/mL (R > 0.99, and the detection limit was 0.008 ng/mL. Compared with ELISA, this method increased the sensitivity by more than two orders of magnitude (the sensitivity of ELISA for p24 is about 1 ng/mL. This immunosensor may be broadly applied to clinical samples, being distinguished by its ease of use, mild reaction conditions, guaranteed reproducibility, and good anti-interference ability.

  6. Immunometric Antibody Sandwich Enzyme-Linked Immunosorbent Assay.

    Science.gov (United States)

    Kohl, Thomas O; Ascoli, Carl A

    2017-06-01

    The antibody sandwich enzyme-linked immunosorbent assay (ELISA) is the most commonly used assay for rapid and accurate detection of antigens. It displays greater sensitivity compared with the indirect ELISA and can be used to determine absolute antigen concentrations in unknown samples provided purified antigen standards are available, although it requires the use of two different antibodies. Briefly, wells are coated with antigen-specific capture antibody then incubated with samples containing unknown antigen. Washing removes unbound antigen and exogenous sample protein before incubation with a second antigen-specific detection antibody, washing, and reincubation with a reporter-labeled tertiary antibody. After tertiary antibody is washed off, substrate is added and hydrolysis is measured spectrophotometrically. The signal intensity is directly proportional to the concentration of the antigen in the test sample. © 2017 Cold Spring Harbor Laboratory Press.

  7. Fabrication of sandwich-type MgB{sub 2}/Boron/MgB{sub 2} Josephson junctions with rapid annealing method

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Song; Wang, Xu; Ma, Junli; Cui, Ruirui; Deng, Chaoyong, E-mail: cydeng@gzu.edu.cn

    2015-11-15

    Sandwich-type MgB{sub 2}/Boron/MgB{sub 2} Josephson junctions were fabricated using magnetron sputtering system. The rapid-anneal process was adopted to replace traditional way of annealing, trying to solve the problem of interdiffusion and oxidation with multilayer films. The boron film was used as barrier layer to avoid the introduction of impurities and improve reproducibility of the junctions. The bottom MgB{sub 2} thin films deposited on c-plane sapphire substrate exhibits a critical temperature T{sub C} of 37.5 K and critical current density J{sub C} at 5 K of 8.7 × 10{sup 6} A cm{sup −2}. From the XRD pattern, the bottom MgB{sub 2} thin film shows c-axis orientation, whereas the top MgB{sub 2} became polycrystalline as Boron barrier layer grown thicker. Therefore, all junction samples show lower T{sub C} than single MgB{sub 2} thin film. The junctions exhibit excellent quasiparticle characteristics with ideal dependence on temperature and Boron barrier thickness. Subharmonic gap structure was appeared in conductance characteristics, which was attributed to the multiple Andreev reflections (MAR). The result demonstrates great promise of this new fabrication technology for MgB{sub 2} Josephson junction fabrication. - Highlights: • Sandwich-type MgB{sub 2}/Boron/MgB{sub 2} Josephson junctions were fabricated. • The junctions were annealed after deposition with the rapid-anneal process. • The highest critical current is 25.3 mA at 5 K and remains non-zero near 25 K. • Subharmonic gap features can be observed in the dI/dV – V curves.

  8. Development and evaluation of a DAS-ELISA for rapid detection of Tembusu virus using monoclonal antibodies against the envelope protein.

    Directory of Open Access Journals (Sweden)

    Hao Chen

    Full Text Available Since April 2010, Tembusu virus (TMUV which is a contagious pathogen of waterfowls, causing symptoms of high fever, loss of appetite and fall in egg production, has been reported in east of China. A double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA which detects for TMUV was developed, using two monoclonal antibodies (mAbs against the TMUV envelope (E protein. BALB/c mice were immunized with purified recombinant E protein expressed in E. coli. Three hybridoma cell lines designated as 12B1, 10C6 and 2D2, were screened by cell fusion and indirect ELISA for their ability to recognize different linear epitopes on the E protein, and were characterized subsequently. High-affinity mAbs 12B1 and 2D2 were used as capture and detection antibodies, respectively. The reaction conditions for the DAS-ELISA were optimized for TMUV detection. The cross-reactivity of the DAS-ELISA was determined using TMUV, duck plague virus, avian influenza virus subtype H9, Newcastle disease virus, duck hepatitis A virus type 1 and duck reovirus samples. A total of 191 homogenized tissues of field samples were simultaneously detected by DAS-ELISA and by RT-PCR. The former was found to have a high specificity of 99.1% and a sensitivity of 93.1%. These results reveal a positive coincidence between DAS-ELISA and RT-PCR at a coincidence rate of 95.8%. The method developed in this study can be used for the diagnosis of TMUV infection of duck origin.

  9. Sandwich-dot enzyme-linked immunosorbent assay for the detection of canine distemper virus.

    Science.gov (United States)

    Li, Zhi; Zhang, Yanlong; Wang, Huiguo; Jin, Jinhua; Li, Wenzhe

    2013-10-01

    A sandwich-dot enzyme-linked immunosorbent assay (dot ELISA) was developed for the detection of canine distemper virus (CDV). In 56 dogs suspected to have CD the rates of detection of CDV antigen in samples of blood lymphocytes and palpebral conjunctiva by dot ELISA and ELISA were, respectively, 91% (49/54) and 81% (44/54) for the lymphocyte samples and 88% (28/32) and 75% (24/32) for the conjunctival samples. The CDV detection limits were 10 ng/50 μL for dot ELISA and 40 ng/50 μL for ELISA. The reliability of dot ELISA relative to electron microscopy was 96% with 22 samples: all 21 samples in which CDV particles were observed by electron microscopy yielded positive results with dot ELISA; the single sample in which particles were not observed yielded false-positive results with dot ELISA. The results indicate that the dot ELISA developed can serve as a reliable rapid diagnostic test in suspected cases of CD and also be useful for epidemiologic surveillance of the disease.

  10. ELISA与RFFIT两种方法检测人狂犬病抗体的对比%Comparison of Two Methods of ELISA and RFFIT for Detection of Human Rabies Antibody

    Institute of Scientific and Technical Information of China (English)

    熊春

    2016-01-01

    Objective To compare enzyme linked immunosorbent assay (ELISA) and rapid fluorescent focus inhibition test (RFFIT) on the detection of human rabies antibodies.Methods100 cases of serum of rabies antibody vaccine recipients after vaccination were selected and randomly detected with RFFIT or ELISA. Results 0 d,3 d,7 d,14 d and 45 d after vaccination,the positive detection rate by ELISA were respectively 9.00%,12.00%,25.00%, 48.00% and 86.00%. The positive rate by RFFIT were 0.00%,0.00%,23.00%,100.00% and 100.00%.ConclusionBoth ELISA and RFFIT can be applied to the detection of rabies virus antibody,but ELISA has higher false positive rate in early period of immunization and higher false negative rate in later period of immunization.%目的:对比酶联免疫吸附试验(ELISA)与快速免疫荧光灶抑制试验(RFFIT)检测人狂犬病抗体的效果。方法选取狂犬抗体疫苗接种者免疫后血清100份,分别随机选用ELISA与RFFIT检测。结果免疫0 d、3 d、7 d、14 d和45 d时ELISA法检测阳性率分别为9.00%、12.00%、25.00%、48.00%、86.00%;RFFIT法检测阳性率分别为0.00%、0.00%、23.00%、100.00%、100.00%。结论 ELISA与RFFIT两种方法检测人狂犬病抗体均可适用于抗狂犬病毒抗体检测,但ELISA法免疫早期假阳性率较高,后期假阴性率较高。

  11. Comparative Diagnosis of Serum IgG1 and Coproantigen ELISA for Fasciolosis Detection of Goats in Mexico

    Science.gov (United States)

    Molina-Mendoza, Pedro; Hernández-Guzmán, Karina; Olivares-Pérez, Jaime; Sarracent-Pérez, Jorge; Zumaquero-Ríos, José

    2016-01-01

    The objective of present study was to determine the prevalence of natural caprine fasciolosis in the Mixteca region of Mexico using coproantigen and serum IgG1 ELISA tests for comparative purposes. A total of 1070 serum and faecal samples were analyzed for IgG1 antibodies and coproantigens, using ELISA with E/S products as antigen and a monoclonal antibody-based sandwich ELISA. Prevalence of 73.46% was found using the serological ELISA and a percentage of 77.20 was found for coproantigen ELISA. The diagnostic sensitivity and specificity for serum ELISA were 86.7% and 96.4%, and for the coproantigen ELISA they were 93.1% and 97.8%, respectively. The seropositive samples were further categorized as low, medium, or high positivity. Results show a great proportion of low and medium positive goats when the serum ELISA test was used. Correlation coefficients between coproantigens and seropositivity were statistically significant (P < 0.01) for low seropositivity (r = 0.93) and medium seropositivity (r = 0.84). The accuracy of faecal antigen ELISA was higher compared to indirect ELISA serological test. Two ELISAs were shown to be useful for demonstrating the current status of F. hepatica infection in the endemic areas and can be employed in studies on epidemiology as well as anthelmintics treatment for preventing economic loss and the risk of transmission to humans. PMID:27563665

  12. Comparative Diagnosis of Serum IgG1 and Coproantigen ELISA for Fasciolosis Detection of Goats in Mexico.

    Science.gov (United States)

    Villa-Mancera, Abel; Molina-Mendoza, Pedro; Hernández-Guzmán, Karina; Olivares-Pérez, Jaime; Sarracent-Pérez, Jorge; Zumaquero-Ríos, José

    2016-01-01

    The objective of present study was to determine the prevalence of natural caprine fasciolosis in the Mixteca region of Mexico using coproantigen and serum IgG1 ELISA tests for comparative purposes. A total of 1070 serum and faecal samples were analyzed for IgG1 antibodies and coproantigens, using ELISA with E/S products as antigen and a monoclonal antibody-based sandwich ELISA. Prevalence of 73.46% was found using the serological ELISA and a percentage of 77.20 was found for coproantigen ELISA. The diagnostic sensitivity and specificity for serum ELISA were 86.7% and 96.4%, and for the coproantigen ELISA they were 93.1% and 97.8%, respectively. The seropositive samples were further categorized as low, medium, or high positivity. Results show a great proportion of low and medium positive goats when the serum ELISA test was used. Correlation coefficients between coproantigens and seropositivity were statistically significant (P < 0.01) for low seropositivity (r = 0.93) and medium seropositivity (r = 0.84). The accuracy of faecal antigen ELISA was higher compared to indirect ELISA serological test. Two ELISAs were shown to be useful for demonstrating the current status of F. hepatica infection in the endemic areas and can be employed in studies on epidemiology as well as anthelmintics treatment for preventing economic loss and the risk of transmission to humans.

  13. Comparative Diagnosis of Serum IgG1 and Coproantigen ELISA for Fasciolosis Detection of Goats in Mexico

    Directory of Open Access Journals (Sweden)

    Abel Villa-Mancera

    2016-01-01

    Full Text Available The objective of present study was to determine the prevalence of natural caprine fasciolosis in the Mixteca region of Mexico using coproantigen and serum IgG1 ELISA tests for comparative purposes. A total of 1070 serum and faecal samples were analyzed for IgG1 antibodies and coproantigens, using ELISA with E/S products as antigen and a monoclonal antibody-based sandwich ELISA. Prevalence of 73.46% was found using the serological ELISA and a percentage of 77.20 was found for coproantigen ELISA. The diagnostic sensitivity and specificity for serum ELISA were 86.7% and 96.4%, and for the coproantigen ELISA they were 93.1% and 97.8%, respectively. The seropositive samples were further categorized as low, medium, or high positivity. Results show a great proportion of low and medium positive goats when the serum ELISA test was used. Correlation coefficients between coproantigens and seropositivity were statistically significant (P<0.01 for low seropositivity (r=0.93 and medium seropositivity (r=0.84. The accuracy of faecal antigen ELISA was higher compared to indirect ELISA serological test. Two ELISAs were shown to be useful for demonstrating the current status of F. hepatica infection in the endemic areas and can be employed in studies on epidemiology as well as anthelmintics treatment for preventing economic loss and the risk of transmission to humans.

  14. DETERMINATION OF HISTAMINE IN FISH USING ELISA TECHNIQUE

    NARCIS (Netherlands)

    KRUGER, C; SEWING, U; STENGEL, G; KEMA, [No Value; WESTERMANN, J; MANZ, B

    1995-01-01

    The analysis of histamine in fish and fish products via competitive ELISA is described. The advantages of this method are easy sample preparation and handling, screening capabilities, and low costs. Automation enables the performance of the assay with higher series of samples. The Histamine-ELISA is

  15. Detection of Potato Leaf Roll Virus (PLRV), Potato Virus Y (PVY) and Potato Virus X (PVX) on Five Potato Varieties by Using of DAS-ELISA and RT-PCR Methods

    OpenAIRE

    Kuswinanti, Tutik

    2012-01-01

    Potato is a staple food crop that widely grown around the world. Virus infection is main factor that affects great loss of the potato production. Potato virus X(PVX), potato virus Y(PVY),and potato leaf roll virus(PLRV) are top three viruses that result in decreased yield of potato in Indonesia. Therefore, the rapid methods of DAS-ELISA was studied to test tuber samples of five potato varieties, Granola, Atlantik, Raja, Super John, Kalosi, and Masalle. Two simple, rapid, sensitive, reliable...

  16. Modelling of Debond and Crack Propagation in Sandwich Structures Using Fracture and Damage Mechanics

    DEFF Research Database (Denmark)

    Berggreen, C.; Simonsen, Bo Cerup; Toernqvist, Rikard

    2003-01-01

    Skin-core de-bonding or core crack propagation will often be dominating mechanisms in the collapse modes of sandwich structures. This paper presents two different methods for prediction of crack propagation in a sandwich structure: a fracture mechanics approach, where a new mode-mix method...

  17. Serotype assignment by sero-agglutination, ELISA, and PCR

    Science.gov (United States)

    For assessing isolates of Listeria monocytogenes serotype designation is the foremost subtyping method used. Traditionally serotyping has been done with agglutination reactions. In the last decade alternative serotyping methods were described using Enzyme Linked Immunosorbent Assay(ELISA)and Polymer...

  18. Immunometric Double-Antibody Sandwich Enzyme-Linked Immunosorbent Assay.

    Science.gov (United States)

    Kohl, Thomas O; Ascoli, Carl A

    2017-06-01

    The double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) is preferentially used to determine the concentration of unknown antibody in a sample. Pure antigen is not required in this assay; however, the use of a reporter-labeled detection antibody is essential. The double-antibody sandwich ELISA is suitable for epitope mapping of different monoclonal antibodies that have been generated against a single antigen. First, plates are coated with a capture antibody specific for immunoglobulins generated by immunization of a host species. Next, the test antibody solution (e.g., serum) is incubated with the capture antibody to facilitate binding. The plates are washed to remove unbound antibody, and then antigen is added. The plates are washed again followed by the addition of an antigen-specific reporter-labeled antibody. Following incubation, unbound reporter antibody is washed off, and reporter-specific substrate is added. Reporter-mediated substrate hydrolysis is visualized and measured. The signal is proportional to the number of test antibodies present in the serum. © 2017 Cold Spring Harbor Laboratory Press.

  19. Neurofilament ELISA validation.

    NARCIS (Netherlands)

    Petzold, A.; Altintas, A.; Andreoni, L.; Bartos, A.; Berthele, A.; Blankenstein, M.A.; Buee, L.; Castellazzi, M.; Cepok, S.; Comabella, M.; Constantinescu, C.S.; Deisenhammer, F.; Deniz, G.; Erten, G.; Espino, M.; Fainardi, E.; Franciotta, D.; Freedman, M.S.; Giedraitis, V.; Gilhus, N.E.; Giovannoni, G.; Glabinski, A.; Grieb, P.; Hartung, H.P.; Hemmer, B.; Herukka, S.K.; Hintzen, R.; Ingelsson, M.; Jackson, S.; Jacobsen, S.; Jafari, N.; Jalosinski, M.; Jarius, S.; Kapaki, E.; Kieseier, B.C.; Koel-Simmelink, M.J.; Kornhuber, J.; Kuhle, J.; Kurzepa, J.; Lalive, P.H.; Lannfelt, L.; Lehmensiek, V.; Lewczuk, P.; Livrea, P.; Marnetto, F.; Martino, D.; Menge, T.; Norgren, N.; Papuc, E.; Paraskevas, G.P.; Pirttila, T.; Rajda, C.; Rejdak, K.; Ricny, J.; Ripova, D.; Rosengren, L.; Ruggieri, M.; Schraen, S.; Shaw, G.; Sindic, C.; Siva, A.; Stigbrand, T.; Stonebridge, I.; Topcular, B.; Trojano, M.; Tumani, H.; Twaalfhoven, H.A.; Vecsei, L.; Pesch, V. Van; Stichele, H. van der; Vedeler, C.; Verbeek, M.M.; Villar, L.M.; Weissert, R.; Wildemann, B.; Yang, C.; Yao, K.; Teunissen, C.E.

    2010-01-01

    BACKGROUND: Neurofilament proteins (Nf) are highly specific biomarkers for neuronal death and axonal degeneration. As these markers become more widely used, an inter-laboratory validation study is required to identify assay criteria for high quality performance. METHODS: The UmanDiagnostics NF-light

  20. Neurofilament ELISA validation.

    NARCIS (Netherlands)

    Petzold, A.; Altintas, A.; Andreoni, L.; Bartos, A.; Berthele, A.; Blankenstein, M.A.; Buee, L.; Castellazzi, M.; Cepok, S.; Comabella, M.; Constantinescu, C.S.; Deisenhammer, F.; Deniz, G.; Erten, G.; Espino, M.; Fainardi, E.; Franciotta, D.; Freedman, M.S.; Giedraitis, V.; Gilhus, N.E.; Giovannoni, G.; Glabinski, A.; Grieb, P.; Hartung, H.P.; Hemmer, B.; Herukka, S.K.; Hintzen, R.; Ingelsson, M.; Jackson, S.; Jacobsen, S.; Jafari, N.; Jalosinski, M.; Jarius, S.; Kapaki, E.; Kieseier, B.C.; Koel-Simmelink, M.J.; Kornhuber, J.; Kuhle, J.; Kurzepa, J.; Lalive, P.H.; Lannfelt, L.; Lehmensiek, V.; Lewczuk, P.; Livrea, P.; Marnetto, F.; Martino, D.; Menge, T.; Norgren, N.; Papuc, E.; Paraskevas, G.P.; Pirttila, T.; Rajda, C.; Rejdak, K.; Ricny, J.; Ripova, D.; Rosengren, L.; Ruggieri, M.; Schraen, S.; Shaw, G.; Sindic, C.; Siva, A.; Stigbrand, T.; Stonebridge, I.; Topcular, B.; Trojano, M.; Tumani, H.; Twaalfhoven, H.A.; Vecsei, L.; Pesch, V. Van; Stichele, H. van der; Vedeler, C.; Verbeek, M.M.; Villar, L.M.; Weissert, R.; Wildemann, B.; Yang, C.; Yao, K.; Teunissen, C.E.

    2010-01-01

    BACKGROUND: Neurofilament proteins (Nf) are highly specific biomarkers for neuronal death and axonal degeneration. As these markers become more widely used, an inter-laboratory validation study is required to identify assay criteria for high quality performance. METHODS: The UmanDiagnostics NF-light

  1. 1/3 SUBHARMONIC SOLUTION OF ELLIPTICAL SANDWICH PLATES

    Institute of Scientific and Technical Information of China (English)

    李银山; 张年梅; 杨桂通

    2003-01-01

    The problem of nonlinear forced oscillations for elliptical sandwich plates is dealtwith. Based on the governing equations expressed in terms of five displacement components,the nonlinear dynamic equation of an elliptical sandwich plate under a harmonic force isderived. A superpositive-iterative harmonic balance ( SIHB ) method is presented for thesteady-state analysis of strongly nonlinear oscillators. In a periodic oscillation, the periodicsolutions can be expressed in the form of basic harmonics and bifurcate harmonics. Thus,an oscillation system which is described as a second order ordinary differential equation,can be expressed as fundamental differential equation with fundamental harmonics andincremental differential equation with derived harmonics. The 1/3 subharnonic solution ofan elliptical sandwich plate is investigated by using the methods of SIHB. The SIHB methodis compared with the numerical integration method. Finally, asymptotical stability of the1/3 subharmonic oscillations is inspected.

  2. Development of an Ultrasensitive ELISA-Bienzyme Colorimetric Substrate Recycle Assay for Measurement of Tau

    Institute of Scientific and Technical Information of China (English)

    DUAN Qiu-hong; WANG Xiao-chuan; WANG Xi-ming; ZHOU Xin-wen; HE Shan-shu; WANG Jian-zhi

    2005-01-01

    On the basis of conventional enzyme-linked immunosorbent assay (ELISA) and bienzyme substrate recycle, ELISA-bienzyme colorimetric substrate recycle was developed in the present study. The sensitivity of this method increased 15 times than that of ELISA for the measurement of Tau and increased 55 times for p-Tau. The linear detective rang of this method expanded 3 times higher than that of conventional ELISA for Tau and had the same as ELISA for p-Tau. So, ELISA-bienzyme colorimetric substrate recycle could be used to detect abnormally phosphorylated tau in cerebrospinal fluid.

  3. Technology sandwich panels with mineral wool insulation

    OpenAIRE

    Tyulenev M.; Burtzeva M.; Mednikova E.

    2016-01-01

    Sandwich panel — self–supporting structure consisting of metal cladding and thermal insulation core. As a heat–insulating core used mineral wool, foamed plastics. Production of sandwich panels with insulation mineral wool performed on modular lines for the production of aggregate or conveyer scheme. Sandwich panels are used as load–bearing elements of the facades, as well as a roof covering.

  4. The feasibility of harmonizing gluten ELISA measurements.

    Science.gov (United States)

    Rzychon, Malgorzata; Brohée, Marcel; Cordeiro, Fernando; Haraszi, Reka; Ulberth, Franz; O'Connor, Gavin

    2017-11-01

    Many publications have highlighted that routine ELISA methods do not give rise to equivalent gluten content measurement results. In this study, we assess this variation between results and its likely impact on the enforcement of the EU gluten-free legislation. This study systematically examines the feasibility of harmonizing gluten ELISA assays by the introduction of: a common extraction procedure; a common calibrator, such as a pure gluten extract and an incurred matrix material. The comparability of measurements is limited by a weak correlation between kit results caused by differences in the selectivity of the methods. This lack of correlation produces bias that cannot be corrected by using reference materials alone. The use of a common calibrator reduced the between-assay variability to some extent, but variation due to differences in selectivity of the assays was unaffected. Consensus on robust markers and their conversion to "gluten content" are required. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  5. Critical velocity of sandwich cylindrical shell under moving internal pressure

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Critical velocity of an infinite long sandwich shell under moving internal pres-sure is studied using the sandwich shell theory and elastodynamics theory. Propagation of axisymmetric free harmonic waves in the sandwich shell is studied using the sandwich shell theory by considering compressibility and transverse shear deformation of the core, and transverse shear deformation of face sheets. Based on the elastodynamics theory, displacement components expanded by Legendre polynomials, and position-dependent elastic constants and densities are introduced into the equations of motion. Critical ve-locity is the minimum phase velocity on the desperation relation curve obtained by using the two methods. Numerical examples and the finite element (FE) simulations are pre-sented. The results show that the two critical velocities agree well with each other, and two desperation relation curves agree well with each other when the wave number κ is relatively small. However, two limit phase velocities approach to the shear wave velocities of the face sheet and the core respectively when k limits to infinite. The two methods are efficient in the investigation of wave propagation in a sandwich cylindrical shell when κ is relatively small. The critical velocity predicted in the FE simulations agrees with theoretical prediction.

  6. Design of sandwich acoustic window for sonar domes

    Institute of Scientific and Technical Information of China (English)

    YU Mengsa; LI Dongsheng; GONG Li; XU Jian

    2005-01-01

    Aimed at the low noise design of sonar dome in ships, a method has been presented for calculating the sonar self noise of a simplified sonar dome consisting of sandwich acoustic window and parallel acoustic cavity, which is excited by stationary random pressure fluctuation of turbulence boundary layer, using temporal and spatial double Fourier transform and wavenumber-frequency spectrum analysis. After numerically analyzing the influence of geometrical and physical parameters of acoustic window on the sonar self noise, the design method and reasonable parameters for sandwich acoustic window are proposed. The results show that the property of low noise induced by acoustic window of sandwich is dominated by the cut-off effect of longitudinal wave and transverse wave propagating in the visco-elastic layer of sandwich as well as the mismatch effect of impedance. If the thickness, density, Young's modulus and damping factor of plates and visco-elastic layer as well as the sound speed of longitudinal wave and transverse wave in the visco-elastic layer are selected reasonably, the maximum noise reduction of sandwich acoustic window is 6.5 dB greater than that of a single glass fiber reinforced plastic plate.

  7. Wire and Packing Tape Sandwiches

    Science.gov (United States)

    Rabinowitz, Sandy

    2009-01-01

    In this article, the author describes how students can combine craft wire with clear packing tape to create a two-dimensional design that can be bent and twisted to create a three-dimensional form. Students sandwich wire designs between two layers of tape. (Contains 1 online resource.)

  8. Thermal-Diode Sandwich Panel

    Science.gov (United States)

    Basiulis, A.

    1986-01-01

    Thermal diode sandwich panel transfers heat in one direction, but when heat load reversed, switches off and acts as thermal insulator. Proposed to control temperature in spacecraft and in supersonic missiles to protect internal electronics. In combination with conventional heat pipes, used in solar panels and other heat-sensitive systems.

  9. High temperature structural sandwich panels

    Science.gov (United States)

    Papakonstantinou, Christos G.

    High strength composites are being used for making lightweight structural panels that are being employed in aerospace, naval and automotive structures. Recently, there is renewed interest in use of these panels. The major problem of most commercial available sandwich panels is the fire resistance. A recently developed inorganic matrix is investigated for use in cases where fire and high temperature resistance are necessary. The focus of this dissertation is the development of a fireproof composite structural system. Sandwich panels made with polysialate matrices have an excellent potential for use in applications where exposure to high temperatures or fire is a concern. Commercial available sandwich panels will soften and lose nearly all of their compressive strength temperatures lower than 400°C. This dissertation consists of the state of the art, the experimental investigation and the analytical modeling. The state of the art covers the performance of existing high temperature composites, sandwich panels and reinforced concrete beams strengthened with Fiber Reinforced Polymers (FRP). The experimental part consists of four major components: (i) Development of a fireproof syntactic foam with maximum specific strength, (ii) Development of a lightweight syntactic foam based on polystyrene spheres, (iii) Development of the composite system for the skins. The variables are the skin thickness, modulus of elasticity of skin and high temperature resistance, and (iv) Experimental evaluation of the flexural behavior of sandwich panels. Analytical modeling consists of a model for the flexural behavior of lightweight sandwich panels, and a model for deflection calculations of reinforced concrete beams strengthened with FRP subjected to fatigue loading. The experimental and analytical results show that sandwich panels made with polysialate matrices and ceramic spheres do not lose their load bearing capability during severe fire exposure, where temperatures reach several

  10. Development of a new method for the determination of residues of the neonictinoid insecticide imidacloprid in juvenile Chinook (Oncorhynchus tyshawytscha) using ELISA detection

    Science.gov (United States)

    Frew, John A.; Grue, Christian E.

    2012-01-01

    The neonicotinoid insecticide imidacloprid (IMI) has been proposed as an alternative to carbaryl for controlling indigenous burrowing shrimp on commercial oyster beds in Willapa Bay and Grays Harbor, Washington. A focus of concern over the use of this insecticide in an aquatic environment is the potential for adverse effects from exposure to non-target species residing in the Bay, such as juvenile Chinook (Oncorhynchus tshawytscha) and cutthroat trout (O. clarki). Federal registration and State permiting approval for the use of IMI will require confirmation that the compound does not adversely impact these salmonids following field applications. This will necessitate an environmental monitoring program for evaluating exposure in salmonids following the treatment of beds. Quantification of IMI residues in tissue can be used for determining salmonid exposure to the insecticide. Refinement of an existing protocol using liquid-chromatography mass spectrometry (LC-MS) detection would provide the low limits of quantification, given the relatively small tissue sample sizes, necessary for determining exposure in individual fish. Such an approach would not be viable for the environmental monitoring effort in Willapa Bay and Grays Harbor due to the high costs associated with running multiple analyses, however. A new sample preparation protocol was developed for use with a commercially available enzyme-linked immunosorbent assay (ELISA) for the quantification of IMI, thereby providing a low-cost alternative to LC-MS for environmental monitoring in Willapa Bay and Grays Harbor. Extraction of the analyte from the salmonid brain tissue was achieved by Dounce homogenization in 4.0 mL of 20.0 mM Triton X-100, followed by a 6 h incubation at 50–55 °C. Centrifugal ultrafiltration and reversed phase solid phase extraction were used for sample cleanup. The limit of quantification for an average 77.0 mg whole brain sample was calculated at 18.2 μg kg-1 (ppb) with an average

  11. Development of a new method for the determination of residues of the neonicotinoid insecticide imidacloprid in juvenile chinook (Oncorhynchus tshawytscha) using ELISA detection.

    Science.gov (United States)

    Frew, John A; Grue, Christian E

    2012-03-01

    The neonicotinoid insecticide imidacloprid (IMI) has been proposed as an alternative to carbaryl for controlling indigenous burrowing shrimp on commercial oyster beds in Willapa Bay and Grays Harbor, Washington. A focus of concern over the use of this insecticide in an aquatic environment is the potential for adverse effects from exposure to non-target species residing in the Bay, such as juvenile Chinook (Oncorhynchus tshawytscha) and cutthroat trout (O. clarki). Federal registration and State permiting approval for the use of IMI will require confirmation that the compound does not adversely impact these salmonids following field applications. This will necessitate an environmental monitoring program for evaluating exposure in salmonids following the treatment of beds. Quantification of IMI residues in tissue can be used for determining salmonid exposure to the insecticide. Refinement of an existing protocol using liquid-chromatography mass spectrometry (LC-MS) detection would provide the low limits of quantification, given the relatively small tissue sample sizes, necessary for determining exposure in individual fish. Such an approach would not be viable for the environmental monitoring effort in Willapa Bay and Grays Harbor due to the high costs associated with running multiple analyses, however. A new sample preparation protocol was developed for use with a commercially available enzyme-linked immunosorbent assay (ELISA) for the quantification of IMI, thereby providing a low-cost alternative to LC-MS for environmental monitoring in Willapa Bay and Grays Harbor. Extraction of the analyte from the salmonid brain tissue was achieved by Dounce homogenization in 4.0 mL of 20.0 mM Triton X-100, followed by a 6 h incubation at 50-55 °C. Centrifugal ultrafiltration and reversed phase solid phase extraction were used for sample cleanup. The limit of quantification for an average 77.0 mg whole brain sample was calculated at 18.2 μg kg(-1) (ppb) with an average

  12. NONLINEAR VIBRATION OF CIRCULAR SANDWICH PLATES UNDER CIRCUMJACENT LOAD

    Institute of Scientific and Technical Information of China (English)

    DU Guo-jun; MA Jian-qing

    2006-01-01

    Based on yon Karman plate theory, the issue about nonlinear vibration for circular sandwich plates under circumjacent load with the loosely clamped boundary condition was researched. Nonlinear differential eigenvalue equations and boundary conditions of the problem were formulated by variational method and then their exact static solution can be got. The solution was derived by modified iteration method, so the anslytic relations between amplitude and nonlinear oscillating frequency for circular sandwich plates were obtained. When circumjacent load makes the lowest natural frequency zero,critical load is obtained.

  13. Buckling analysis of sandwich plate using layer wise theory

    Energy Technology Data Exchange (ETDEWEB)

    Ranjbaran, Arash; Khoshravan, Mohammad Reza [University of Tabriz, Tabriz (Iran, Islamic Republic of); Kharazi, Mahsa [Sahand University of Technology, Sahand (Iran, Islamic Republic of)

    2014-07-15

    Buckling analysis of sandwich plate was investigated using layer wise method. The formulation was based on the first-order shear deformation theory, and the Rayleigh-Ritz method was used for approximating and determining the displacement field. The results obtained from layer wise theory was compared with finite element results and showed good agreement. This study demonstrated that layer wise theory could describe buckling behavior of sandwich plates with high accuracy and represents a more realistic and acceptable description of behavior of the plates with much less computational cost.

  14. Study on voids of epoxy matrix composites sandwich structure parts

    Science.gov (United States)

    He, Simin; Wen, Youyi; Yu, Wenjun; Liu, Hong; Yue, Cheng; Bao, Jing

    2017-03-01

    Void is the most common tiny defect of composite materials. Porosity is closely related to composite structure property. The voids forming behaviour in the composites sandwich structural parts with the carbon fiber reinforced epoxy resin skins was researched by adjusting the manufacturing process parameters. The composites laminate with different porosities were prepared with the different process parameter. The ultrasonic non-destructive measurement method for the porosity was developed and verified through microscopic examination. The analysis results show that compaction pressure during the manufacturing process had influence on the porosity in the laminate area. Increasing the compaction pressure and compaction time will reduce the porosity of the laminates. The bond-line between honeycomb core and carbon fiber reinforced epoxy resin skins were also analyzed through microscopic examination. The mechanical properties of sandwich structure composites were studied. The optimization process parameters and porosity ultrasonic measurement method for composites sandwich structure have been applied to the production of the composite parts.

  15. Establishment of optimized ELISA system specific for HLA-G in body fluids.

    Science.gov (United States)

    Ouji-Sageshima, N; Geraghty, D E; Ishitani, A; Hatake, K; Ito, T

    2016-12-01

    Recently, human leukocyte antigen-G (HLA-G) has been a focus in the field of reproductive immunology, tumor progression and transplantation, because of its inhibitory function as ligand to the inhibitory receptors leukocyte immunoglobulin-like receptors (LILR) B1 and LILRB2. The HLA-G is expressed in distinct mRNA isoforms, one of which encodes a soluble HLA-G (sHLA-G) protein, detectable by sandwich ELISA. Therefore, sHLA-G ELISAs have been used as a noninvasive diagnosis system. While a number of sHLA-G-specific ELISAs have been described, our prior studies showed that data obtained by the conventional ELISA system detecting sHLA-G in body fluids was not consistent with the data obtained from immunoprecipitation (IP)/immunoblotting (IB). Therefore, we established an optimized ELISA system described in this report, which yields results consistent with IP/IB analysis. Using this system, we determined sHLA-G protein in amniotic fluids, and found that sHLA-G levels at preterm (∼36 weeks) were clearly higher than those at term (37-41 weeks). These data and supporting experiments showed that the ELISA system we established can be an useful tools for the detection of sHLA-G protein in body fluids than the conventional ELISA system.

  16. Robust and Air-Stable Sandwiched Organo-Lead Halide Perovskites for Photodetector Applications

    KAUST Repository

    Mohammed, Omar F.

    2016-02-25

    We report the simplest possible method to date for fabricating robust, air-stable, sandwiched perovskite photodetectors. Our proposed sandwiched structure is devoid of electron or hole transporting layers and also the expensive electrodes. These simpler architectures may have application in the perovskite-only class of solar cells scaling up towards commercialization.

  17. Quantitative determination of VEGF165 in cell culture medium by aptamer sandwich based chemiluminescence assay.

    Science.gov (United States)

    Shan, Siwen; He, Ziyi; Mao, Sifeng; Jie, Mingsha; Yi, Linglu; Lin, Jin-Ming

    2017-08-15

    In this work, we have developed a sensitive and selective chemiluminescence (CL) assay for vascular endothelial growth factor (VEGF165) quantitative detection based on two specific VEGF165 binding aptamers (Apt). VEGF is a predominant biomarker in cancer angiogenesis, and sensitive detection method of VEGF are highly demanded for both academic study and clinical diagnosis of multiple cancers. In our experiment, VEGF165 was captured in a sandwich structure assembled by two binding aptamers, one capture aptamer was immobilized on streptavidin-coated magnetic beads (MBs) and another VEGF-binding aptamer was labeled by biotin for further phosphatase conjunction. After Apt-VEGF-Apt sandwich was formed on MBs surface, alkaline phosphatase (ALP) was modified to the second aptamer to catalyze CL reaction. By applying 4-methoxy-4-(3-phosphatephenyl)-spiro-(1,2-dioxetane-3,2-adamantane) (AMPPD) as CL substrate, strong signal intensity was achieved. VEGF165 content as low as 1ng/mL was detected in standard spiked samples by our assay, and linear range of working curve was confirmed from 1 to 20ng/mL. Then our method was successfully applied for cell culture medium analysis and on-chip hypoxic HepG2-HUVEC co-culture model study with excellent accuracy equal to ELISA Kit. Our developed assay demonstrated an outstanding performance in VEGF165 quantification and may be further extended to clinical testing of important biomarkers as well as probing microchip cell culture model. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Clinical Value of ELISPOT and ELISA Method in Diagnosis of Tuberculosis%酶联免疫斑点法和ELISA法诊断结核病临床价值的探讨

    Institute of Scientific and Technical Information of China (English)

    石海萍; 周晓芹

    2016-01-01

    目的:探讨酶联免疫斑点法(ELISPOT)和ELISA法诊断结核病的临床价值。方法:86例疑似结核病患者,同时行ELISPOT和ELISA法的检验,对结果做配对卡方检验及Kappa一致性检验,比较两种方法的检验的结果,探讨临床诊断应用的价值。结果:配对卡方检验,P值为0.774,>0.05,不拒绝H0,二者无差异。Kappa一致性检验,Kappa值为0.579,0.4<Kappa值≤0.75表示一致性中等。结论:两种方法检验结果差异无显着性,具有一致性,结合实验操作性,方便性,ELISA法更适合应用于临床。%Objective: To investigate the clinical value of enzyme linked immunosorbent assay (ELISPOT) and ELISA method in the diagnosis of tuberculosis.Methods:86 cases of suspected tuberculosis patients. At the same time, the ELISPOT and ELISA test, the paired chi square test and kappa consistency test, to compare the two methods of test results of the value of clinical application.Results:Paired chi square test,P value was 0.774,>0.05, does not reject H0, no difference between the two. Kappa consistency test, Kappa = 0.579, Kappa>0.4 or lower than 0.75 medium consistency.Conclusion:There were no significant differences between the two methods. The results showed that the ELISA method was more suitable for clinical application.

  19. Experimental study on mechanical properties of aircraft honeycomb sandwich structures

    Directory of Open Access Journals (Sweden)

    Talebi Mazraehshahi H.

    2010-06-01

    Full Text Available Mechanical behaviour of sandwich panels under different conditions have been exprimentally studied in this research to increase the knowledge of aircraft sandwich panel structures and facilitate design criteria for aircraft structures. Tests were concentrated on the honeycomb sandwich structures under different loads including flexural, insert shear, flat wise tension and compression loads. Furthermore, effect of core density and face material on mechanical behavior of different samples were investigated and compared with analytical and FEM method. Effects of skin thickness on strength of honycomb sandwhich panels under shear pull out and moments have also been considerd in this study. According to this investigation, insert strength and flexural test under different load conditions is strongly affected by face thickness, but compression and tearoff (falt wise tensile properties of a sandwich panel depends on core material. The study concludes that the correlation between experimental results and the analytical predictions will enable the designer to predict the mechanical behaviour and strength of a sandwich beam; however, applied formula may lead engineers to unreliable results for shear modulus.

  20. Thermal buckling analysis of truss-core sandwich plates

    Institute of Scientific and Technical Information of China (English)

    陈继伟; 刘咏泉; 刘伟; 苏先樾

    2013-01-01

    Truss-core sandwich plates have received much attention in virtue of the high values of strength-to-weight and stiffness-to-weight as well as the great ability of impulse-resistance recently. It is necessary to study the stability of sandwich panels under the influence of the thermal load. However, the sandwich plates are such complex three-dimensional (3D) systems that direct analytical solutions do not exist, and the finite element method (FEM) cannot represent the relationship between structural parameters and mechanical properties well. In this paper, an equivalent homogeneous continuous plate is idealized by obtaining the effective bending and transverse shear stiffness based on the characteristics of periodically distributed unit cells. The first order shear deformation theory for plates is used to derive the stability equation. The buckling temperature of a simply supported sandwich plate is given and verified by the FEM. The effect of related parameters on mechanical properties is investigated. The geometric parameters of the unit cell are optimized to attain the maximum buckling temperature. It is shown that the optimized sandwich plate can improve the resistance to thermal buckling significantly.

  1. Comparative Analysis of Human Growth Hormone in Serum Using SPRi, Nano-SPRi and ELISA Assays.

    Science.gov (United States)

    Vance, Stephen; Zeidan, Effat; Henrich, Vincent C; Sandros, Marinella G

    2016-01-07

    Sensitive and selective methods for the detection of human growth hormone (hGH) over a wide range of concentrations (high levels of 50-100 ng ml(-) (1) and minimum levels of 0.03 ng ml(-) (1)) in circulating blood are essential as variable levels may indicate altered physiology. For example, growth disorders occurring in childhood can be diagnosed by measuring levels of hGH in blood. Also, the misuse of recombinant hGH in sports not only poses an ethical issue it also presents serious health threats to the abuser. One popular strategy for measuring hGH misuse, relies on the detection of the ratio of 22 kDa hGH to total hGH, as non-22 kDa endogenous levels drop after exogenous recombinant hGH (rhGH) administration. Surface plasmon resonance imaging (SPRi) is an analytical tool that allows direct (label-free) monitoring and visualization of biomolecular interactions by recording changes of the refractive index adjacent to the sensor surface in real time. In contrast, the most frequently used colorimetric method, enzyme-linked immunosorbent assay (ELISA) uses enzyme labeled detection antibodies to indirectly measure analyte concentration after the addition of a substrate that induces a color change. To increase detection sensitivity, amplified SPRi uses a sandwich assay format and near infrared quantum dots (QDs) to increase signal strength. After direct SPRi detection of recombinant rhGH in spiked human serum, the SPRi signal is amplified by the sequential injection of detection antibody coated with near-infrared QDs (Nano-SPRi). In this study, the diagnostic potential of direct and amplified SPRi was assessed for measuring rhGH spiked in human serum and compared directly with the capabilities of a commercially available ELISA kit.

  2. The difference of ELISA and LABScreen in detecting HLA antibodies

    Institute of Scientific and Technical Information of China (English)

    吴萍萍

    2013-01-01

    Objective To compare the difference of ELISA and LABScreen in detecting HLA antibodies and evaluate their effects on allograft rejection.Methods Consecutive patients undergoing kidney transplantion from November,2008 to December,2009 in the First Affiliated Hospital

  3. Extended high order sandwich panel theory for bending analysis of sandwich beams with carbon nanotube reinforced face sheets

    Science.gov (United States)

    Jedari Salami, S.

    2016-02-01

    Bending analysis of a sandwich beam with soft core and carbon nanotube reinforced composite (CNTRC) face sheets in the literature is presented based on Extended High order Sandwich Panel Theory (EHSAPT). Distribution of fibers through the thickness of the face sheets could be uniform or functionally graded (FG). In this theory the face sheets follow the first order shear deformation theory (FSDT). Besides, the two dimensional elasticity is used for the core. The field equations are derived via the Ritz based solution which is suitable for any essential boundary condition. The influences of boundary conditions on bending response of the sandwich panel with soft core and CNTRC face sheet are investigated. In each type of boundary condition the effect of distribution pattern of CNTRCs on many essential involved parameters of the sandwich beam with functionally graded carbon nanotube reinforced composite (FG- CNTRC) face sheets are studied in detail. Finally, experimental result have been compared with those obtained based on developed solution method. It is concluded that, the sandwich beam with X distribution figure of face sheets is the strongest with the smallest transverse displacement, and followed by the UD, O and ∧-ones, respectively.

  4. Free vibration Analysis of Sandwich Plates with cutout

    Science.gov (United States)

    Mishra, N.; Basa, B.; Sarangi, S. K.

    2016-09-01

    This paper presents the free vibration analysis of sandwich plates with cutouts. Cutouts are inevitable in structural applications and the presence of these cutouts in the structures greatly influences their dynamic characteristics. A finite element model has been developed here using the ANSYS 15.0 software to study the free vibration characteristics of sandwich plates in the presence of cutouts. Shell 281 element, an 8-noded element with six degrees of freedom suited for analyzing thin to moderately thick structures is considered in the development of the model. Block Lanczose method is adopted to extract the mode shapes to obtain the natural frequency corresponding to free vibration of the plate. The effects of parametric variation on the natural frequency of the sandwich plates with cutout are studied and results are presented.

  5. Vibration analysis and optimization of sandwich composite with curvilinear fibers

    Science.gov (United States)

    Honda, S.; Narita, Y.

    2016-09-01

    The present paper develops a shell element based on the refined zigzag theory (RZT) and applies it to the vibration analysis and optimization problem of the composite sandwich plate composed of CFRP skins and soft-cores. The RZT accepts large differences in layer stiffness, and requires less calculation effort than the layer-wise or three-dimensional theories. Numerical results revealed that the present method predicts vibration characteristics of composite sandwich plates with soft-core accurately. Then, shapes of reinforcing fibers in CFRP composite skins are optimized to maximize fundamental frequencies. As an optimizer, the particle swarm optimization (PSO) approach is employed since curvilinear fiber shapes are defined by continuous design variables. Obtained results showed that the composite sandwich with optimum curvilinear fiber shapes indicates higher fundamental frequencies compared with straight fibers.

  6. 基于匹配追踪的蜂窝夹层复合材料损伤检测%Research on honeycomb sandwich composite structure damage detection based on matching pursuit method

    Institute of Scientific and Technical Information of China (English)

    冯勇明; 周丽; 李真

    2012-01-01

    基于Lamb波和匹配追踪时频分析方法,提出一种损伤成像方法,对蜂窝夹层复合材料结构进行损伤监测.首先针对Lamb波传播的特点,提出了匹配追踪方法的快速实现方案,该方法能准确地匹配失真变形的窄带脉冲信号,并识别Lamb波的模态;然后对由压电传感器采集到的Lamb波信号,采用匹配追踪方法提取特征信息,得到Lamb波的能量分布;在此基础上,考虑Lamb波在各向异性结构中传播速度的影响,将损伤处的散射波能量分布和各像素点对比度联系起来,得到损伤图像,将损伤的情况可视化.通过蜂窝夹层复合材料结构实验验证了该方法的可行性和有效性.%This study proposes a damage imaging method using Lamb wave and matching pursuit method time-frequency analysis for damage detection of honeycomb sandwich structure. Matching pursuit method is employed to decompose Lamb wave signals into a linear expansion of several chirplet atoms using a fast realization algorithm. The relationship between Lamb wave' s dispersion and the chirplet' s chirp rate is established, which can be used to identify the modes of Lamb waves. Then the matching pursuit method is applied to the Lamb wave signals excited and sensed by piezoelectric sensors in the time-frequency domain, which can obtain the energy distribution of the scattered waves. Considering the effect of anisotropic property on the velocity distributions of Lamb waves, the damage image can be obtained by the time-dependent energy distribution of scattered waves. The effectiveness and accuracy of the proposed method in identifying the modes and in locating defects are demonstrated by the experimental results on the honeycomb sandwich composite structure.

  7. Assessing hazelnut allergens by protein- and DNA-based approaches: LC-MS/MS, ELISA and real-time PCR.

    Science.gov (United States)

    Costa, Joana; Ansari, Parisa; Mafra, Isabel; Oliveira, M Beatriz P P; Baumgartner, Sabine

    2014-04-01

    Hazelnut (Corylus avellana L.) is responsible for a significant part of the allergies related to nuts. Still, it is a very much appreciated nut and as consequence is widely used in all types of processed foods, such as chocolates. Correct food labelling is currently the most effective means of preventing the consumption of allergenic ingredients, namely hazelnut, by the sensitised/allergic individuals. Thus, to verify labelling compliance and to ensure allergic patient protection, the development of highly sensitive methodologies is of extreme importance. In this study, three major methodologies, namely enzyme-linked immunosorbent assays (ELISA), liquid chromatography coupled with mass spectrometry and real-time polymerase chain reaction, were evaluated for their performance regarding the detection of hazelnut allergens in model chocolates. The sandwich ELISA and respective antibodies were in-house developed and produced. With sensitivity levels of approximately 1 mg kg(-1) and limits of quantification of 50-100 mg kg(-1), all the performed methods were considered appropriate for the identification of hazelnut in complex foods such as chocolates. To our knowledge, this was the first successful attempt to develop and compare three independent approaches for the detection of allergens in foods.

  8. A New Silicon-Based Ferroelectric Sandwich Structure

    Institute of Scientific and Technical Information of China (English)

    任天令; 张林涛; 刘理天; 李志坚

    2001-01-01

    A new silicon-based PbTiO3/Pb(Zr0.53 Ti0.47)O3/PbTiO3 sandwich structure is fabricated by a sol-gel method. Compared with other fabrication processes without PbTiO3 buffer layers, the annealing temperature is greatly reduced by as much as 100℃. Capacitance-voltage, polarization-electric field and dielectric-frequency properties of this sandwich structure are studied. The Pb(Zrx Ti1-x)O3 films are proved to have good dielectric and ferroelectric properties.

  9. New-sandwiched hydrogen separation membranes with low environmental impact

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Y.; Ozaki, T.; Komaki, M.; Nishimura, C. [Eco-Energy Materials Group, Ecomaterials Center, Ibaraki (Japan)

    2003-07-01

    There is a strong interest in economical methods for hydrogen separation. In this work, we have developed sandwiched membranes, Pd-Ag/V-15Ni/Pd-Ag, which had much lower environmental impact than palladium-based membranes, which are the most commonly used membrane for the current production of highly pure hydrogen. The Pd-Ag overlayer was prepared on V-15Ni substrates (thickness 40 {mu}m) using multi-target sputtering. The hydrogen permeation of the Pd-Ag/V-15Ni/Pd-Ag sandwiched membranes was examined using gas permeation technology. (orig.)

  10. Analysis of fenbendazole residues in bovine milk by ELISA.

    Science.gov (United States)

    Brandon, David L; Bates, Anne H; Binder, Ronald G; Montague, William C; Whitehand, Linda C; Barker, Steven A

    2002-10-09

    Fenbendazole residues in bovine milk were analyzed by ELISAs using two monoclonal antibodies. One monoclonal antibody (MAb 587) bound the major benzimidazole anthelmintic drugs, including fenbendazole, oxfendazole, and fenbendazole sulfone. The other (MAb 591) was more specific for fenbendazole, with 13% cross-reactivity with the sulfone and no significant binding to the sulfoxide metabolite. The limit of detection of the ELISA method in the milk matrix was 7 ppb for MAb 587 and 3 ppb for MAb 591. Fenbendazole was administered in feed, drench, and paste form to three groups of dairy cattle. Milk was collected immediately before dosing and then every 12 h for 5 days. The ELISA indicated that residue levels varied widely among individual cows in each group. Fenbendazole levels peaked at approximately 12-24 h and declined rapidly thereafter. Metabolites were detected at much higher levels than the parent compound, peaked at approximately 24-36 h, and declined gradually. Residue levels were undetectable by 72 h. The ELISA data correlated well with the total residues determined by chromatographic analysis, but the use of the two separate ELISAs did not afford an advantage over ELISA with the single, broadly reactive MAb 587. The ELISA method could be used to flag high-residue samples in on-site monitoring of fenbendazole in milk and is a potential tool for studying drug pharmacokinetics.

  11. Establishment of recombinant major allergens Bet v 1 and Phl p 5a as Ph. Eur. reference standards and validation of ELISA methods for their measurement. Results from feasibility studies.

    Science.gov (United States)

    Vieths, S; Barber, D; Chapman, M; Costanzo, A; Daas, A; Fiebig, H; Hanschmann, K M; Hrabina, M; Kaul, S; Ledesma, A; Moingeon, P; Reese, G; Schörner, C; van Ree, R; Weber, B; Buchheit, K H

    2012-04-01

    The potency of allergen extracts is determined as total allergenic activity without consideration of their composition and the units differ from one manufacturer to another, making it very difficult to compare the different products. Recently, purified major allergens have been obtained by recombinant DNA technology and produced under Good Manufacturing Practice (GMP) conditions. In principle, such recombinant allergens could be established as reference standards and could help for the standardisation of the major allergen content of allergen extracts. Two recombinant major allergens, one from birch pollen, rBet v 1, and one from Timothy grass pollen, Phl p 5a, have been selected at the end of the CREATE programme as a potential starting point for the establishment as European Pharmacopoeia (Ph. Eur.) Reference Standards through a project run by the Biological Standardisation Programme (BSP) of the European Directorate for the Quality of Medicines & HealthCare (EDQM). To this end, bulk candidate recombinant materials, produced under GMP conditions, were procured from two European manufacturers and subsequently formulated and lyophilised. Four ELISA systems from three different manufacturers were included in the project, two for Bet v 1 and two for Phl p 5a with the aim of establishing reference methods for determination of the respective major antigens both in natural allergen extracts as well as in recombinant allergen products. The project was run in 3 phases: a preparatory and preliminary testing phase (feasibility phase or Phase 1), an extended feasibility phase carried out in 3 laboratories (Phase 2) to confirm the transferability of the methods and an international collaborative study with a large number of participating laboratories (Phase 3). This article describes the work done in Phase 1 and Phase 2, i.e. the physico-chemical and biological characterisation of the recombinant candidate reference standards, the assessment of their suitability for the

  12. Determination of abscisic acid and its glucosyl ester in embryogenic callus cultures of Vitis vinifera in relation to the maturation of somatic embryos using a new liquid chromatography-ELISA analysis method.

    Science.gov (United States)

    Prado, María Jesús; Largo, Asier; Domínguez, Cristina; González, María Victoria; Rey, Manuel; Centeno, María Luz

    2014-06-15

    The levels of abscisic acid (ABA), its conjugate ABA-GE, and IAA were determined in embryogenic calli of Vitis vinifera L. (cv. Mencía) cultured in DM1 differentiation medium, to relate them to the maturation process of somatic embryos. To achieve this goal, we developed an analytical method that included two steps of solid-phase extraction, chromatographic separation by HPLC, ABA-GE hydrolysis, and sensitive ELISA quantification. Because the ABA immunoassay was based on new polyclonal antibodies raised against a C4'-ABA conjugate, the assay was characterized (detection limit, midrange, measure range, and cross-reaction) and validated by a comparison of the ABA data obtained with this ELISA procedure and with a physicochemical method (LC-ESI-MS/MS). Radioactive-labeled internal standards were initially added to callus extracts to correct the losses of plant hormones, and thus assure the accuracy of the measurements. The endogenous concentration of ABA in the embryogenic callus cultured in DM1 medium was doubled at the fifth week of culture, concurring with the maturation process of somatic embryos, as indicated by the accumulation of carbohydrates observed through histological analysis. The ABA-GE content was higher than ABA, decreasing at 21 days of culture in DM1 medium but increasing thereafter. The data suggest the involvement of the synthesis and conjugation of ABA in the final stages of development in grapevine somatic embryos from embryogenic callus. IAA levels were low, suggesting that auxin plays no significant role during the maturation of somatic embryos. In addition, the lower ABA levels in calli cultured in DM differentiation medium with PGRs, a medium presenting high precocious germination and deficiencies in somatic embryo development indicate that an increase in ABA content during the development of somatic embryos in grapevine is necessary for their correct maturation. Copyright © 2014 Elsevier GmbH. All rights reserved.

  13. Kinetic ELISA in Microfluidic Channels

    Directory of Open Access Journals (Sweden)

    Debashis Dutta

    2011-06-01

    Full Text Available In this article, we describe the kinetic ELISA of Blue Tongue and Epizootic Hemorrhagic Disease viral antibodies in microfluidic channels by monitoring the rate of generation of the enzyme reaction product under static conditions. It has been shown that this format of the immunoassay allows very reliable quantitation of the target species using inexpensive glass microchips and a standard epifluorescence microscope system coupled to a CCD camera. For the viral antibodies assayed here, the limit of detection (LOD for the analyte concentration in our microchips was established to be 3–5 times lower than that obtained on commercial microwell plates using a fiftieth of the sample volume and less than a third of the incubation time. Our analyses further show that when compared to the end-point ELISA format, the kinetic mode of this assay yields an improvement in the LOD by over an order of magnitude in microfluidic devices. This benefit is primarily realized as the observed variation in the background fluorescence (signal at the start of the enzyme reaction period was significantly larger than that in the rate of signal generation upon repeating these assays in different microchannels/microchips. Because the kinetic ELISA results depend only on the latter quantity, the noise level in them was substantially lower compared to that in its end-point counterpart in which the absolute fluorescence measurements are of greater significance. While a similar benefit was also recorded through implementation of kinetic ELISAs on the microwell platform, the improvement in LOD registered in that system was not as significant as was observed in the case of microfluidic assays.

  14. Plasmonic ELISA for the ultrasensitive detection of Treponema pallidum.

    Science.gov (United States)

    Nie, Xin-Min; Huang, Rong; Dong, Cai-Xia; Tang, Li-Juan; Gui, Rong; Jiang, Jian-Hui

    2014-08-15

    In this report, we have developed a plasmonic ELISA strategy for the detection of syphilis. Plasmonic ELISA is an enzyme-linked immunoassay combined with enzyme-mediated surface plasmon resonance (SPR) of gold nanoparticles (AuNPs). Immune response of the Treponema pallidum (T. pallidum) antibodies triggers the acetylcholinesterase-catalyzed hydrolysis of acetylthiocholine to produce abundant thiocholine. The positive charged thiol, in turn, alters the surface charge distribution the AuNPs and leads to the agglomeration of the AuNPs. The induced strong localized SPR effect of the agglomerate AuNPs can, thus, allow the quantitative assay of T. pallidum antibodies due to the remarkable color and absorption spectral response changes of the reaction system. The plasmonic ELISA exhibited a quasilinear response to the logarithmic T. pallidum antibody concentrations in the range of 1pg/mL-10ng/mL with a detection limit of 0.98pg/mL. Such a low detection limit was 1000-fold improvements in sensitivity over a conventional ELISA. The results of plasmonic ELISA in syphilis assays of serum specimens from 60 patients agreed with those obtained using a conventional ELISA method. The plasmonic ELISA has characteristics (analyte specific, cost-effective, ease of automatic, low limit of detection) that provide potential for diagnosis and therapeutic monitoring of syphilis.

  15. Evaluation of a fast and simple sample preparation method for PBDE flame retardants and DDT pesticides in fish for analysis by ELISA compared with GC-MS/MS

    Science.gov (United States)

    A simple, fast, and cost-effective sample preparation method, previously developed and validated for the analysis of organic contaminants in fish using low-pressure gas chromatography tandem mass spectrometry (LPGC-MS/MS), was evaluated for analysis of polybrominated diphenyl ethers (PBDEs) and dich...

  16. [Evaluation of a new ELISA (Bartels) for detection of Legionella pneumophila antigen in urine].

    Science.gov (United States)

    de Ory, Fernando

    2002-03-01

    Detection of Legionella pneumophila soluble antigens allows rapid diagnosis of pneumonia caused by these bacteria. A new ELISA (Bartels) for antigenuria detection has recently been commercialized. We compared the new ELISA with another well-established ELISA (Binax). To evaluate ELISA-Bartels (Legionella Urinary Antigen, Intracel, Issaquah, Washington, United States), urine samples previously characterized by ELISA Binax (Legionella Urinary Antigen Enzyme Immunoassay Kit, Binax, Portland, Maine, United States) were used. Samples came from Legionella outbreaks (n = 48), from sporadic legionellosis (n = 38), and from children with viral pneumonia (n = 21). Samples from the External Quality Control of Legionella of the European Working Group on Legionella Infections (n = 102) were also tested. Of the samples analyzed, 109 were positive in ELISA-Binax, 2 were equivocal and 98 were negative. Samples showing equivocal results were excluded from the analysis. The sensitivity of ELISA-Bartels in comparison with that of ELISA-Binax was 98.2% (107/109) and specificity was 82.7% (81/98). In the 17 samples that were positive in ELISA-Bartels and negative in ELISA-Binax, 10 were positive in ELISA-Binax after concentration by selective ultrafiltration and 6 further cases showed serology indicating or compatible with recent Legionella infection and were thus classified as true positives. ELISA-Bartels showed good sensitivity and specificity. Sensitivity was even higher than that of ELISA-Binax. Thus, we consider it to be an appropriate method for diagnosis of Legionella pneumonia.

  17. Universal quantum dot-based sandwich-like immunoassay strategy for rapid and ultrasensitive detection of small molecules using portable and reusable optofluidic nano-biosensing platform

    Energy Technology Data Exchange (ETDEWEB)

    Zhou, Liping; Zhu, Anna; Lou, Xuening; Song, Dan; Yang, Rong [School of Environment and Natural Resources, Renmin University of China, Beijing (China); Shi, Hanchang [School of Environment, Tsinghua University, Beijing (China); Long, Feng, E-mail: longf04@ruc.edu.cn [School of Environment and Natural Resources, Renmin University of China, Beijing (China)

    2016-01-28

    A universal sandwich-like immunoassay strategy based on quantum-dots immunoprobe (QD-labeled anti-mouse IgG antibody) was developed for rapid and ultrasensitive detection of small molecules. A portable and reusable optofluidic nano-biosensing platform was applied to investigate the sandwich-like immunoassay mechanism and format of small molecules, as well as the binding kinetics between QD immunoprobe and anti-small molecule antibody. A two-step immunoassay method that involves pre-incubation mixture of different concentration of small molecule and anti-small molecule antibody, and subsequent introduction of QD immunoprobe into the optofluidic cell was conducted for small molecule determination. Compared with the one-step immunoassay method, the two-step immunoassay method can obtain higher fluorescence signal and higher sensitivity index, thus improving the nano-biosensing performance. Based on the proposed strategy, two mode targets, namely, microcystin-LR (MC-LR) and Bisphenol A (BPA) were tested with high sensitivity, rapidity, and ease of use. A higher concentration of small molecules in the sample led to less anti-small molecule antibody bound with antigen-carrier protein conjugate immobilized onto the sensor surface, and less QD immunoprobes bound with anti-small molecule antibody. This phenomenon lowered the fluorescence signal detected by nano-biosensing platform. Under optimal operating conditions, MC-LR and BPA exhibited a limit of detection of 0.003 and 0.04 μg/L, respectively. The LODs were better than those of the indirect competitive immunoassay method for small molecules via Cy5.5-labeled anti-small molecule antibody. The proposed QD-based sandwich-like immunoassay strategy was evaluated in spiked water samples, and showed good recovery, precision and accuracy without complicated sample pretreatments. All these results demonstrate that the new detection strategy could be readily applied to the other trace small molecules in real water samples

  18. Diagnostic accuracy of ELISA methods as an alternative screening test to indirect immunofluorescence for the detection of antinuclear antibodies. Evaluation of five commercial kits.

    Science.gov (United States)

    Tonuttia, Elio; Bassetti, Danila; Piazza, Anna; Visentini, Daniela; Poletto, Monica; Bassetto, Franca; Caciagli, Patrizio; Villalta, Danilo; Tozzoli, Renato; Bizzaro, Nicola

    2004-03-01

    Detection of antinuclear antibodies (ANA) is a fundamental laboratory test for diagnosing systemic autoimmune diseases. Currently, the method of choice is indirect immunofluorescence (IIF) on a HEp-2 cell substrate. The goal of this study was to evaluate the diagnostic accuracy of five commercially available enzyme immunoassay (EIA) kits for ANA detection and to verify the possibility of using them as an alternative to the IIF method. The study involved 1513 patients, 315 of whom were diagnosed with a systemic autoimmune disease and 1198 in whom an autoimmune disorder was excluded. For all sera, ANA detection was performed via IIF and with five different EIA kits. The results were evaluated in relation to clinical diagnosis and the presence of possible specific autoantibodies (anti-ENA or anti-dsDNA); lastly, they were compared with the results obtained using ANA-IIF as the method of reference. The positive rate of the ANA-IIF test in subjects with systemic autoimmune diseases was 92%, whereas in the five ANA-EIA kits there was broad diversity in terms of response, with positive rates ranging from 74 to 94%. All the EIA kits correctly detected the presence of antibodies (anti-dsDNA, anti-RNP, anti-Ro/SSA) responsible for homogeneous and speckled fluorescence pattern, but at the same time they showed substantial inaccuracy with the nucleolar pattern, with a mean sensitivity of approximately 50% in this case. Instead, there was a large kit-to-kit difference in terms of identification of anti-Scl70 and centromere patterns, for which sensitivities ranged between 45 and 91%, and between 49 and 100%, respectively. The results of the study demonstrate that the commercially available ANA-EIA kits show different levels of sensitivity and specificity. Some of them have a diagnostic accuracy that is comparable and, in some cases, even higher than the IIF method. Consequently, these could be used as an alternative screening test to IIE. However, others do not ensure acceptable

  19. Bending and Deformation of Sandwich Panels Due to Localized Pressure

    Directory of Open Access Journals (Sweden)

    Bambang K. Hadi

    2005-05-01

    Full Text Available Bending and deformation of sandwich panels due to localized pressure were analyzed using both Rayleigh-Ritz and finite element methods. The faces were made of laminated composite plates, while the core was a honeycomb material. Carbon fiber and glass fiber reinforced plastics were used for composite plate faces. In the case of Rayleigh-Ritz method, first the total energy of the system was calculated and then taking the variations of the total energy, the sandwich panel deflections could be computed. The deflections were assumed by means of Fourier series. A finite element code NASTRAN was exploited extensively in the finite element method. 3-dimensional 8-node brick elements were used to model sandwich panels, for both the faces sheets and the core. The results were then compared to each other and in general they are in good agreements. Dimple phenomena were found in these cases. It shows that localized pressure on sandwich structures will produce dimple on the pressurize region with little effects on the rest of the structures.

  20. Detection of botulinum neurotoxin serotype B at sub mouse LD(50 levels by a sandwich immunoassay and its application to toxin detection in milk.

    Directory of Open Access Journals (Sweden)

    Miles C Scotcher

    Full Text Available BACKGROUND: Botulinum neurotoxin (BoNT, the causative agent of botulism, a serious neuroparylatic disease, is produced by the anaerobic bacterium Clostridium botulinum and consists of a family of seven serotypes (A-H. We previously reported production of high-affinity monoclonal antibodies to BoNT serotype A. METHODS AND FINDINGS: Recombinant peptide fragments of the light chain, the transmembrane and receptor-binding domains of the heavy chain of botulinum neurotoxin type B (BoNT/B were expressed in Escherichia coli as GST-fusion proteins and purified. These proteins were used to immunize BALB/cJ mice for the generation of monoclonal antibodies (mAbs. Antibody-producing hybridomas were detected using either a direct binding ELISA binding to plate-immobilized BoNT/B, or with a capture-capture ELISA whereby the capacity of the antibody to capture BoNT/B from solution was tested. A total of five mAbs were selected, two of which bound the toxin light chain and three bound the receptor-binding domain of BoNT/B heavy chain. MAb MCS6-27 was identified via capture-capture ELISA and was the only mAb able to bind BoNT/B in solution under physiological conditions. MAbs F24-1, F26-16, F27-33 and F29-40 were identified via direct binding ELISA, and were able to capture BoNT/B in solution only in the presence of 0.5-0.9 mM sodium dodecyl sulphate (SDS. MAb MCS6-27 and an anti-BoNT/B polyclonal antibody were incorporated into a sandwich ELISA that did not require SDS. CONCLUSIONS: We report here the generation of monoclonal antibodies to serotype B and the subsequent development of a sensitive sandwich immunoassay. This immunoassay has a detection limit of 100 fg BoNT/B, fifty times more sensitive than the mouse bioassay detection limit of 5 pg BoNT/B. Additionally, this assay detected as little as 39 pg/mL of toxin in skim, 2% and whole milk.

  1. Development of a sandwich enzyme-linked immunosorbent assay for dTMP-GH fusion protein by rational immunogen selection.

    Science.gov (United States)

    Wang, Song; Shen, Mingqiang; Chen, Shilei; Wang, Cheng; Chen, Fang; Chen, Mo; Zhao, Gaomei; Ran, Xinze; Cheng, Tianmin; Su, Yongping; Xu, Yang; Wang, Junping

    2017-12-01

    dTMP-GH is a chimeric protein containing a tandem dimer of thrombopoietin mimetic peptide (dTMP) fused to human growth hormone (hGH) prepared previously by our team. It shows significant bioactivity in promoting thrombocytopoiesis, but detection of intact dTMP-GH in plasma is still a challenge due to the presence of endogenous hGH. In this study, a rabbit polyclonal antibody with high affinity to dTMP was obtained with a BSA-conjugated immunogen composed of 20 amino acids sequence spanning two TMP and the linker. A monoclonal antibody termed as 3B2 was screened out by using immunizing mice with whole dTMP-GH, which was proved to simultaneously interact with rhGH, TMP-GH, and dTMP-GH, respectively. In this study, we developed a specific and sensitive sandwich enzyme-linked immunosorbent assay (ELISA) with two antibodies (one polyclonal and one HRP-conjugated monoclonal) to quantify dTMP-GH. The polyclonal antibody and HRP-conjugated monoclonal antibody 3B2 were applied as the capture antibody and detection antibody, respectively. A good correlation between ELISA and bicinchoninic acid (BCA) assay in the quantification of diluted dTMP-GH was observed (r(2) = 0.996). Meanwhile, the standard curve of this ELISA method was found in a linear relationship between 0.2 and 10 ng/mL in the presence of rabbit plasma. In vivo experiments demonstrate that the newly developed method is effective to detect dTMP-GH in rabbits, which paves the way for further pharmacokinetic evaluation.

  2. Development of a Quantitative ELISA Method for Total Antibody of Haemophilus influenzae Type b%b型流感嗜血杆菌总抗体ELISA定量检测方法的建立

    Institute of Scientific and Technical Information of China (English)

    李红; 李茂光; 唐静; 梁丽; 李亚南; 何莉; 叶强

    2011-01-01

    Objective To develop and preliminarily verify a quantitative ELISA method for total antibody of Haemophilus influenzae type b (Hib). Methods The coating concentration of antigen, working concentration of enzyme-labeled antibody, linear range of reference serum and substrate were optimized by chessboard titration, based on which the developed method was verified for specificity, accuracy and precision. Results The optimal coating concentration of antigen was 0. 4 μg/ml, and the optimal dilution of enzyme-labeled antibody was 1: 6 000. The optimal linear range of reference serum was 5 ~ 100 ng/ml (R2 = 0.997 6). TMB was selected as substrate. The developed method showed high specificity, accuracy and precision. Conclusion A quantitative ELISA method for total antibody of Hib was successfully developed, which might substitute to the determination of anti-Hib IgG in sera.%目的 建立定量检测血清中b型流感嗜血杆菌(Haemophilus influenzae type b,Hib)总抗体的ELISA方法,并进行初步验证.方法 采用棋盘滴定法确定抗原最佳包被浓度及酶标抗体工作浓度,参考血清的线性范围及适合的底物,并验证方法的特异性、准确性及精密性.结果 最佳抗原包被浓度为0.4 μg/ml;最佳酶标抗体稀释度为1:6000;参考血清的最适线性范围为5~100ng/ml,RZ=0.9976 ;选择TMB作为底物.该方法特异性较好,准确性和精密性较高.结论 已成功建立了定量测定血清中Hib总抗体的ELISA方法,可逐渐替代测定血清中抗Hib IgG的方法.

  3. 血清抗碳酸酐酶Ⅲ抗体ELISA检测方法的建立与初步应用%Establishment and preliminary application of the ELISA method for anti-carbonic anhydrase III antibody detection

    Institute of Scientific and Technical Information of China (English)

    刘辰庚; 王培昌

    2011-01-01

    目的 建立人血清抗碳酸酐酶(CA)Ⅲ抗体的ELISA检测方法,并对系统性红斑狼疮、皮肌炎、糖尿病肾病、高血压肾病患者和健康人群的血清抗CAⅢ抗体水平进行初步调查.方法 使用抗CAⅢ抗体标准品、CAⅢ及相应酶标抗体建立血清抗CAⅢ抗体ELISA检测方法,验证试剂稳定性、标本保存稳定性,并进行精密度、灵敏度、回收率、抗干扰性等方法学评价;各项技术指标均合格后对系统性红斑狼疮、皮肌炎、糖尿病肾病和高血压肾病患者的血清进行抗CAⅢ抗体水平检测.结果 成功建立ELISA检测人血清抗CAⅢ抗体的方法,其批内精密度为6.2%,批间精密度为8.2%,灵敏度为0.025,回收率为106%,且具有较好的抗干扰性、试剂稳定性和标本保存稳定性.系统性红斑狼疮和糖尿病肾病患者的血清抗CAⅢ抗体水平高于健康对照相(P<0.05),阳性率分别为43%和18%.皮肌炎和高血压肾病患者的血清抗CAⅢ抗体水平与健康对照组比较无统计学差异(P>0.05),且未出现阳性结果.结论 使用现有市售试剂进行人血清抗CAⅢ抗体的ELISA检测是可行的,抗CAⅢ抗体可能参与了系统性红斑狼疮和糖尿病肾病的发生发展.%Objective To establish an ELISA method for anti-carbonic anhydrase III (CA III ) antibody detection, and to evaluate the serum level of anti-CA III antibody in normal control group and patients with systemic lupus erythemato-sus, dermatomyositis, 2-type diabetic nephropathy and hypertensive nephropathy respectively. Methods To establish the ELISA method using CA III, anti-CA III antibody and enzyme labeled secondary antibody. To evaluate the stability of the regent and sample, and the sensitivity, stability and anti-interference performance of the ELISA method. To investigate the anti-CA III antibody level in serum of normal control group and patients with systemic lupus erythematosus, dermatomyositis, 2-type

  4. Establishment of competitive ELISA method for determination of P53 protein in mouse serum%竞争性ELISA方法检测小鼠血清中的P53蛋白

    Institute of Scientific and Technical Information of China (English)

    武军华; 魏文清; 贾培媛; 王晨宇; 赵宇; 刘晶; 黄春倩; 王玉霞

    2011-01-01

    Objective To establish a method of competitive ELISA to measure P53 or P53-associated bio-drug level in animal tissues. It could be effective in the research of the metabolism and distribution of bio-reagents based on P53 protein.Methods Horseradish peroxidase,HRP,was used to label monoclonal antibody 3P40 against P53. Based on the specific recognition of P53 and 3P40HRP,a competitive ELISA method was established and its within-day precision was evaluated. The influence of mouse serum on the hinding of P53 and 3P40HRP was determined by comparing the result from PBST control. Results HRP labeled 3P40,3P40HRP could specifically bind with P53 by ELISA with the titer of 1: 200,000. The competitive ELISA method had a good within-day precision and the relative standard deviation (RSD) was lower than 5%. In order to assay the influence of tissue content on the binding of 3P40HRP with P53 ,mouse serum was added to the binding solution. Compared with PBST,mouse serum could affect the binding signal in this assay. Using a standard curve obtained with P53 protein in serum sample , the P53 concentration in the serum of mouse ip injected with P53 protein was measured. Conclusion Monoclonal antibody against P53,3P40 ,was labeled with HRP. Based on P53 protein binding with 3P40HRP and then decreased the amount of 3P40HRP hinding with P53 coated on 96 well plate , a competive ELISA method was established. This method could be well used to measure the P53 concentration in mouse serum.%目的建立动物组织内P53蛋白的免疫检测分析方法,为P53生物药物的代谢分布研究提供检测手段.方法辣根过氧化物酶(HRP)标记抗P53蛋白单克隆抗体3P40,应用标记后的抗体及重组P53蛋白的检测,建立竞争性ELISA检测方法;应用此方法建立血清样品中P53蛋白标准曲线,测定腹腔注射P53后不同时间小鼠血清中P53浓度.结果利用亲和层析纯化的3P40,进行HRP标记,ELISA检测3P40HRP滴度可达1:200000:

  5. 进出口食品中组胺的ELISA快速测定%Rapid Detection of Histamine in Import and Export Food by ELISA Method

    Institute of Scientific and Technical Information of China (English)

    郑海松; 杨小娇

    2011-01-01

    本文建立了一种进出口食品中组胺的酶联免疫快速测定方法。针对目前国内市场常见的Abraxis、r-Biopharm和Neogen三种试剂盒,进行比较选择最优试剂盒,并对酶联免疫分析定量检测组胺残留试剂盒进行选择性(交叉反应)测试、特异性与检测限测试以及对进出口食品样品组胺的回收率和精密度测试。结果显示,选择r-Biopharm试剂盒来检测样品中的组胺准确性较好,其对N-酰基-组胺交叉性为100%,对N-甲基-组胺、5-羟基吲哚乙酸、咪唑乙酸、L-组氨酸、N-甲基咪唑乙酸及血清素等组胺类似物交叉反应都小于1%,进出口食品中红葡萄酒的检测限(LOD)为250μg/kg,牛奶的LOD为100μg/kg,奶酪的LOD为2.5 mg/kg,鱼粉的LOD为100 mg/kg,样品的平均回收率在86.4%~95.6%,相对标准偏差2.7%~3.9%,经五个试验室的回收验证均具有较好的回收率,经HPLC确证假阳性率≤2.5%,表明酶联免疫分析定量法可快速、准确实现对食品中组胺残留量的快速筛选。%To established an import and export food histamine by enzyme-linked immunosorbent fast determination method.According to the current domestic market common Abraxis,r-Biopharm and Neogen three kit,carries on the comparison to choose the optimum kit and enzyme-linked immunosorbent analysis quantitative detection of histamine residual kit for selective(cross reaction) testing,specificity and detection limit testing and samples of import and export food of histamine recovery rate and precision testing.The results showed that choose r-Biopharm kit to test samples of the histamine accuracy is better,which to N-Acyl-histamine cross is 100%,to N-Methyl-histamine,5-Hydroxy-indole-acetic acid,Imidazole acetic acid,L-Histidine,N-Methyl imidazole acetic acid and Serotonin cross reaction are less than 1%.Red wine detection limit(LOD) is 250 mg/kg,milk LOD is 100 mg/kg,cheese LOD is 2.5 mg/kg,fishmeal LOD is 100 mg/kg,the average

  6. A study on an efficient prediction of welding deformation for T-joint laser welding of sandwich panel Part II : Proposal of a method to use shell element model

    Directory of Open Access Journals (Sweden)

    Kim Jae Woong

    2014-06-01

    Full Text Available I-core sandwich panel that has been used more widely is assembled using high power CO₂laser welding. Kim et al. (2013 proposed a circular cone type heat source model for the T-joint laser welding between face plate and core. It can cover the negative defocus which is commonly adopted in T-joint laser welding to provide deeper penetration. In part I, a volumetric heat source model is proposed and it is verified thorough a comparison of melting zone on the cross section with experiment results. The proposed model can be used for heat transfer analysis and thermal elasto-plastic analysis to predict welding deformation that occurs during laser welding. In terms of computational time, since the thermal elasto-plastic analysis using 3D solid elements is quite time consuming, shell element model with multi-layers have been employed instead. However, the conventional layered approach is not appropriate for the application of heat load at T-Joint. This paper, Part II, suggests a new method to arrange different number of layers for face plate and core in order to impose heat load only to the face plate.

  7. A study on an efficient prediction of welding deformation for T-joint laser welding of sandwich panel Part II : Proposal of a method to use shell element model

    Science.gov (United States)

    Kim, Jae Woong; Jang, Beom Seon; Kang, Sung Wook

    2014-06-01

    I-core sandwich panel that has been used more widely is assembled using high power CO-laser welding. Kim et al. (2013) proposed a circular cone type heat source model for the T-joint laser welding between face plate and core. It can cover the negative defocus which is commonly adopted in T-joint laser welding to provide deeper penetration. In part I, a volumetric heat source model is proposed and it is verified thorough a comparison of melting zone on the cross section with experiment results. The proposed model can be used for heat transfer analysis and thermal elasto-plastic analysis to predict welding deformation that occurs during laser welding. In terms of computational time, since the thermal elasto-plastic analysis using 3D solid elements is quite time consuming, shell element model with multi-layers have been employed instead. However, the conventional layered approach is not appropriate for the application of heat load at T-Joint. This paper, Part II, suggests a new method to arrange different number of layers for face plate and core in order to impose heat load only to the face plate.

  8. Canine specific ELISA for coagulation factor VII

    DEFF Research Database (Denmark)

    Knudsen, Tom; Kjelgaard-Hansen, Mads; Tranholm, Mikael;

    2011-01-01

    available to date. In this study, a canine specific ELISA for measurement of FVII:Ag in plasma was developed and validated. The FVII:Ag ELISA correctly diagnosed homozygous and heterozygous hereditary FVII deficiency. Together with activity based assays, such as FVII:C, the FVII:Ag ELISA should be valuable...

  9. Modelling of Debond and Crack Propagation in Sandwich Structures Using Fracture and Damage Mechanics

    DEFF Research Database (Denmark)

    Berggreen, C.; Simonsen, Bo Cerup; Toernqvist, Rikard

    2003-01-01

    Skin-core de-bonding or core crack propagation will often be dominating mechanisms in the collapse modes of sandwich structures. This paper presents two different methods for prediction of crack propagation in a sandwich structure: a fracture mechanics approach, where a new mode-mix method...... is presented, and a local damage mechanics approach. The paper presents a real-life application example, where the superstructure in a vessel pulls the skin off the sandwich deck. The calculations show almost unstable crack growth initially followed by a stabilization, and a nearly linear relation between...

  10. Innovative sandwich concepts open up potential for flat structures. Function-integrated lightweight design; Sandwich-Strukturen fuer den funktionsintegrierten Leichtbau. Fahrzeugleichtbau

    Energy Technology Data Exchange (ETDEWEB)

    Kopp, Gerhard; Friedrich, Horst E. [DLR Deutsches Zentrum fuer Luft- und Raumfahrt e.V., Stuttgart (Germany). Inst. fuer Fahrzeugkonzepte; Kuppinger, Jan [Fraunhofer-Institut fuer Chemische Technologie (ICT), Pfinztal (Germany). Abt. Polymer-Engineering; Henning, Frank [Fraunhofer-Institut fuer Chemische Technologie (ICT), Pfinztal (Germany). Abt. Polymer-Engineering; DLR Deutsches Zentrum fuer Luft- und Raumfahrt e.V., Stuttgart (Germany). Kompetenzzentrum; Karlsruher Innovationsclusters ' Technologien fuer den hybriden Leichtbau' (Germany)

    2009-07-01

    Sandwich structures make it possible to achieve an optimum combination of material, form and functional lightweight design. Vehicle weight is reduced and resources are saved. However, such solutions can only be used when the overall concept is cost-effective. For this reason, various DLR and Fraunhofer institutes are currently working on new and cost-effective sandwich structures within the 'Competence Centre for Automotive Lightweight Technology' to further exploit this construction method for flat structures. (orig.)

  11. Composite Sandwich Technologies Lighten Components

    Science.gov (United States)

    2010-01-01

    Leveraging its private resources with several Small Business Innovation Research (SBIR) contracts with both NASA and the U.S. Department of Defense, WebCore Technologies LLC, of Miamisburg, Ohio, developed a fiber-reinforced foam sandwich panel it calls TYCOR that can be used for a wide variety of industrial and consumer applications. Testing at Glenn Research Center?s Ballistic Impact Facility demonstrated that the technology was able to exhibit excellent damage localization and stiffness during impact. The patented and trademarked material has found use in many demanding applications, including marine, ground transportation, mobile shelters, bridges, and most notably, wind turbines.

  12. Influence of weld stiffness on buckling strength of laser-welded web-core sandwich plates

    OpenAIRE

    Jelovica, Jasmin; Romanoff, Jani; Ehlers, Sören; Varsta, Petri

    2012-01-01

    This paper investigates the influence of weld rotation stiffness on the global bifurcation buckling strength of laser-welded web-core sandwich plates. The study is carried out using two methods, the first is the equivalent single-layer theory approach solved analytically for simply supported plates and numerically for clamped plates. First-order shear deformation theory is used. The second method is the three-dimensional model of a sandwich plate solved with finite element method. Both approa...

  13. Competitive ELISA Method for Determining Haemophilus Influenza Type b Polysaccharide%b型流感嗜血杆菌多糖竞争ELISA检测方法的建立

    Institute of Scientific and Technical Information of China (English)

    江山; 刘威; 兰芳; 廖红梅; 李小波; 张立平; 崔长法

    2013-01-01

    Objective To develop competitive ELISA method for determining the concentration of haemophilus influenza type b polysaccharide (PRP).Method PRP-Ty was set as coating antigen and poliasccharide as competition antigen,comparison were made between the two reactions against antisera of haemophilus influenza type b.The accuracy and precision of the method were validated accordingly.Results The optimal coating concentration of PRP-Ty and serum dilution factor was 1 ∶ 400,and 1 ∶ 40K respectively.Quantitation of PRP concentration was linear from 6.25ug/ml-100ug/ml,with lower limit of detection being 3.13ug/ml.The regression equation was B/Bo =-34.328 [PRP] + 105.03 with RSQ =0.995.The coefficient of variation of intra-assay and inter-assay was 3.4%-6.5% and 8.48% respectively.The recovery rate of PRP in culture medium was 95.7%.Conclusion The competitive ELISA method showed both high accuracy and precision,and thus can be used for detection of haemophilus influenza type b polysaccharide.%目的 建立竞争酶联免疫吸附分析法(Enzyme-Linked Immunosorbent Assay,ELISA),测定b型流感嗜血杆菌多糖(PRP)浓度方法以b型流感嗜血杆菌多糖-酪胺(Ty)为包被抗原,待测多糖为竞争抗原,与优化的抗b型流感嗜血杆菌多糖抗体反应,建立标准曲线,并验证该方法的准确性和精密度.结果 优化后的b型流感嗜血杆菌多糖-酪胺包被浓度为1∶400倍稀释,抗多糖血清稀释度为1∶40K.检测线性范围6.25μg/ml~100μg/ml,最低检测限为3.13μg/ml,回归方程为B/B0=-34.328[PRP]+105.03,线性相关系数为R2 =0.995.批内精密度为3.4%~6.5%,批间精密度为8.48%,测定培养基中b型流感嗜血杆菌多糖的回收率为95.7%.结论 本研究建立的b型流感嗜血杆菌多糖竞争ELISA方法准确性和精密性均较好,可以特异性地检测b型流感嗜血杆菌多糖浓度.

  14. HCV游离抗原BA-ELISA检测方法的临床应用评价%Evaluation on clinical application of HCV free antigen detection methods of BA-ELISA

    Institute of Scientific and Technical Information of China (English)

    龙润乡; 陈红英; 朱祥明; 沈云松; 孙翳; 苏品璨; 王俊; 谢忠平

    2013-01-01

    目的 根据国家相关法规对HCV游离抗原BA-ELISA检测方法进行临床应用效果评价.方法 生物素标记丙型肝炎病毒抗体(HCV-Ab)与辣根过氧化物酶标记亲和素联合应用建立HCV游离抗原BA-ELISA检测法,分别在3个省级医疗卫生机构进行临床试验,同时使用HCV-cAg、HCV-Ab、HCV-RNA荧光定量检测试剂盒进行对比同步检测.结果 临床试剂与HCV-Ab检测结果相比一致性66.67%,特异性100%;与荧光定量PCR相比结果一致性为80.85%、特异性为98.87%;同类市售产品HCV核心抗原检测试剂检测结果与之接近.结论 临床试剂特异性较好,灵敏度有待提高;临床研究试剂比市售同类产品阳性检出率略高,但两种试剂检出率均低于核酸及抗体检测试剂.%OBJECTIVE According to the national relative regulations, to evaluate effect of clinical application of HCV free antigen BA-ELISA detection method. METHODS Used biotin-labeled hepatitis C virus antibody (HCV-Ab) and horseradish peroxidase -conjugated avidin system to establish of free HCV antigen detection methods of BA-ELISA. Respectively in the three provincial-level medical and health institutions for clinical trials, meanwhile HCV-cAg, HCV-Ab, HCV-RNA fluorescence quantitative detection kit synchronous detection were compared. RESULTS Results of clinical study reagent compared with HCV-Ab test results, coincidence rate was 66.67%, specificity 100%; And compared with HCV-PCR, coincidence rate was 80.85%, specificity was 98.87%; The results were analogue by the commercial product HCV-Ag detection reagents and HCV-Ag detection reagents. CONCLUSION Better clinical study reagent specificity, sensitivity need to be improved; clinical study reagents than similar products commercially, positive rate is slightly higher, but both are lower than nucleic acid detection reagent and antibody detection kit.

  15. ELISA检测唾液中乙型肝炎表面抗原方法的优化与评价%Evaluation of modified ELISA method to detect Hepatitis B surface antigen in saliva specimens

    Institute of Scientific and Technical Information of China (English)

    张进

    2013-01-01

    目的探讨优化用于检测唾液中乙型肝炎表面抗原的 ELISA法,并对其进行评价。方法对 ELISA的不同条件:样品类型、加样体积和孵育温度及时间进行优化,筛选最佳条件;并采用3种不同的临界值计算方法,判断最佳临界值。结果不做任何处理的唾液样品可直接用于检测(P=0.100),50μL唾液样品与100、150μL加样体积检测结果比较差异无统计学意义(P=0.070)。采用25℃孵育18 h的优化方法检测的结果和对应血清的检测结果相关性最好,优化后唾液检测乙型肝炎表面抗原方法的特异性和敏感性分别达92.6%和93.6%。结论优化后的 ELISA方法在唾液样品的乙型肝炎表面抗原检测中具有潜在的应用价值。%Objective To evaluated a modified ELISA method for detecting the Hepatitis B surface antigen(HBsAg)in saliva samples.Methods Optimized the different conditions,including sample type,sample volume and incubation condition,to screen the best condition.Three different methods to calculate cut-off value was evaluated to screen the most acceptable.Results The non-processed saliva sample can be used directly for detection(P=0.100).Test results of different sample volume was no significant difference(P=0.070).HBsAg was detected in 44 saliva samples out of 47 paired positive serum specimens and not detected in 63 saliva samples out of 68 matched negative serum samples by the optimized ELISA assay.There was excellent agreement between the results for the serum and saliva specimens and the kappa value was 0.87 for saliva specimens.Using an optimized protocol,the sensitivities and specificities were 93.6% and 92.6%,respectively.Conclusion Our data showed a significant promise for the use of the modified commercial ELISA in saliva sample for Hepatitis B virus infection surveillance.

  16. Mechanical Behavior of CFRP Lattice Core Sandwich Bolted Corner Joints

    Science.gov (United States)

    Zhu, Xiaolei; Liu, Yang; Wang, Yana; Lu, Xiaofeng; Zhu, Lingxue

    2017-02-01

    The lattice core sandwich structures have drawn more attention for the integration of load capacity and multifunctional applications. However, the connection of carbon fibers reinforced polymer composite (CFRP) lattice core sandwich structure hinders its application. In this paper, a typical connection of two lattice core sandwich panels, named as corner joint or L-joint, was investigated by experiment and finite element method (FEM). The mechanical behavior and failure mode of the corner joints were discussed. The results showed that the main deformation pattern and failure mode of the lattice core sandwich bolted corner joints structure were the deformation of metal connector and indentation of the face sheet in the bolt holes. The metal connectors played an important role in bolted corner joints structure. In order to save the calculation resource, a continuum model of pyramid lattice core was used to replace the exact structure. The computation results were consistent with experiment, and the maximum error was 19%. The FEM demonstrated the deflection process of the bolted corner joints structure visually. So the simplified FEM can be used for further analysis of the bolted corner joints structure in engineering.

  17. Elastic-plastic deformation of sandwich rod on elastic basis

    Institute of Scientific and Technical Information of China (English)

    GU Yu

    2008-01-01

    Sandwich composite material possesses advantages of both light weight and high strength.Although the mechanical behaviors of sandwich composite material with the influence of single external environment have been intensively studied,little work has been done in the study of mechanical property,in view of the nonlinear behavior of sandwich composites in the complicated external environments.In this paper,the problem about the bending of the three-layer elastic-plastic rod located on the elastic base,with a compressibly physical nonlinear core,has been studied.The mechanical response of the designed three-layer elements consisting of two bearing layers and a core has been examined.The complicated problem about curving of the three-layer rod located on the elastic base has been solved.The convergence of the proposed method of elastic solutions is examined to convince that the solution is acceptable.The calculated results indicate that the plasticity and physical nonlinearity of materials have a great influence on the deformation of the sandwich rod on the elastic basis.

  18. Bending moment of galvanized iron glass fiber sandwich panel

    Directory of Open Access Journals (Sweden)

    Gurustal Somnath Swamy

    2016-05-01

    Full Text Available The main objective of this project is to prepare a laminated with Galvanized iron thickness fractions, fiber volume fractions and orientation in the layers of GF were fabricated by hand lay-up method and evaluated for their bending moment properties of the sandwich panel using universal testing machine. This paper theoretically calculates the bending behavior of sandwich panel. The recent need to develop a new range of materials has resulted in the development of high performance lightweight composites with excellent properties. Metal– composite systems consist of alternating layers of metal and fiber-reinforced polymer composites which are bonded by an adhesive. Sandwich beams were tested under Air Bending. Stress-strain and stress-displacement were recorded by using AIMIL UTM. The beam face sheets exhibited a softening non-linearity on the bending side. Experimental results were in good agreement with predictions from simple models. On an overall basis, the sandwich panel exhibited better bending moment performance than the monolithic galvanized iron

  19. Effect of microencapsulated phase change material in sandwich panels

    Energy Technology Data Exchange (ETDEWEB)

    Castellon, Cecilia; Medrano, Marc; Roca, Joan; Cabeza, Luisa F. [GREA Innovacio Concurrent, Edifici CREA, Universitat de Lleida, Pere de Cabrera s/n, 25001 Lleida (Spain); Navarro, Maria E.; Fernandez, Ana I. [Departamento de Ciencias de los Materiales e Ingenieria Metalurgica, Universitat de Barcelona, Marti i Franques 1, 08028 Barcelona (Spain); Lazaro, Ana; Zalba, Belen [Instituto de Investigacion en Ingenieria de Aragon, I3A, Grupo de Ingenieria Termica y Sistemas Energeticos (GITSE), Dpto. Ingenieria Mecanica, Area de Maquinas y Motores Termicos, Universidad de Zaragoza, Campus Politecnico Rio Ebro, Edificio ' ' Agustin de Betancourt,' ' Maria de Luna s/n, 50018 Zaragoza (Spain)

    2010-10-15

    Sandwich panels are a good option as building materials, as they offer excellent characteristics in a modular system. The goal of this study was to demonstrate the feasibility of using the microencapsulated PCM (Micronal BASF) in sandwich panels to increase their thermal inertia and to reduce the energy demand of the final buildings. In this paper, to manufacture the sandwich panel with microencapsulated PCM three different methods were tested. In case 1, the PCM was added mixing the microencapsulated PCM with one of the components of the polyurethane. In the other two cases, the PCM was added either a step before (case 2) or a step after (case 3) to the addition of the polyurethane to the metal sheets. The results show that in case 1 the effect of PCM was overlapped by a possible increase in thermal conductivity, but an increase of thermal inertia was found in case 3. In case 2, different results were obtained due to the poor distribution of the PCM. Some samples showed the effect of the PCM (higher thermal inertia), and other samples results were similar to the conventional sandwich panel. In both cases (2 and 3), it is required to industrialize the process to improve the results. (author)

  20. [The systematization of the enzyme-linked immunosorbent assay (=ELISA)].

    Science.gov (United States)

    Gränzer, W

    1989-06-01

    Using the example of the detectability of immunological methods the study shows the considerable influence exerted by this detectability on the quantitative and qualitative parameters and the necessity of a systematization of the methods. This systematization already starts with the product labeling: A sufficient description of antibodies has to contain five qualities. The profitability of the experimental research is improved by optimized product and method systematization. ELISA is presented in a systematized form as a modular system optimizing the comparability of these methods. In order to detect the gain of information transparency obtained by systematization, an ELISA, described in the literature, is presented in the original and in the systematized form.

  1. Standard Terminology of Structural Sandwich Constructions

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    2007-01-01

    1.1 This terminology covers terms necessary for a basic uniform understanding and usage of the language peculiar to structural sandwich constructions. The simplest structural sandwich is a three layered construction formed by bonding a thin layer (facing) to each side of a thick layer (core).

  2. Primary hepatocyte culture in collagen gel mixture and collagen sandwich

    Institute of Scientific and Technical Information of China (English)

    Ying-Jie Wang; Hong-Ling Liu; Hai-Tao Guo; Hong-Wei Wen; Jun Liu

    2004-01-01

    AIM: To explore the methods of hepatocytes culture in a collagen gel mixture or between double layers of collagen sandwich configuration and to examine the functional and cytomorphological characteristics of cultured hepatocytes.METHODS: A two-step collagenase perfusion technique was used to isolate the hepatocytes from Wistar rats or newborn Chinese experimental piglets. The isolated hepatocytes were cultured in a collagen gel mixture or between double layers of collagen sandwich configuration respectively. The former was that rat hepatocytes were mixed with type I rat tail collagen solution till gelled, and the medium was added onto the gel. The latter was that swine hepatocytes were seeded on a plate precoated with collagen gel for 24 h, then another layer of collagen gel was overlaid, resulting in a sandwich configuration. The cytomorphological characteristics, albumin secretion, and LDH-release of the hepatocytes cultured in these two models were examined.RESULTS: Freshly isolated rat hepatocytes were successfully mixed and fixed in collagen gel, and cultured in the gel condition. During the culture period, the urea synthesized and secreted by rat hepatocytes was detected throughout the period. Likewise, newborn experimental piglet hepatocytes were successfully fixed between the double layers of collagen gel, forming a sandwich configuration.Within a week of culture, the albumin secreted by swine hepatocytes was detected by SDS/PAGE analysis. The typical cytomorphological characteristics of the hepatocytes cultured by the above two culture models were found under a phasecontrast microscope. There was little LDH-release during the culture period.CONCLUSION: Both collagen gel mixture and double layers of collagen sandwich configuration can provide cultural conditions much closer to in vivoenvironment, and are helpful for maintaining specific hepatic fiJnctions and cytomorphological characteristics. A collagen gel mixture culture may be more eligible for the

  3. Detection of canine echinococcosis by coproantigen ELISA

    Institute of Scientific and Technical Information of China (English)

    DeS; PanD; BeraAK; SreevatsavaV; DasSK; DasS; RanaT; BandyopadhyayS; BhattacharyaD

    2010-01-01

    Objective:To study the canine echinococcosis by coproantigen ELISA method. Methods:During the present investigation experimental infection was established using evaginated worms of Echinococcus granulosus (E. granulosus). To check cross reactivity two pups were infected with Taenia hydatigena(T. hydatigena). In order to detect the presence of antigen, hyperimmune sera were raised against excretory-secretory products of adult worms E. chinococcus granulosus. Faecal sample collected either from experimentally infected pups or from other sources were heated at 70℃to detect heat stable soluble antigen. Results:Pups harbouring less than 104 worms showed negative results. Samples collected from 14 days onwards from experimentally infected animals harbouring more than 104 worms showed positive value. The maximum positive samples were detected in samples collected from in and around slaughter house and the least number of samples were detected positive maintained by dog squad. Conclusions:The affinity purified IgG exhibited promising results for detection of canine echinococcosis by indirect ELISA.

  4. 抗紫杉醇单克隆抗体的制备及ELISA检测方法的建立%Preparation of monoclonal antibody against taxol and establishment of ELISA detection method

    Institute of Scientific and Technical Information of China (English)

    赵凯; 孙立新; 孙宇石; 周东坡

    2012-01-01

    Objective; To prepare monoclonal antibodies (McAb) against taxol and to establish an indirect enzyme-linked immunosordent assay (ELISA) method for rapidly detecting taxol. Methods; Taxol was coupled with the carrier protein bovine serum, albumin (BSA) to prepare the complete antigen taxol-BSA by succinct anhydride method. The prepared taxol-BSA was identified by ultraviolet scanner and thin layer chromatography (TLC). Blab/c mice were immunized with taxol-BSA. The splenocytes of the immunized Blab/c mice were fused with SP2/0 myeloma cells by PEG 4000, and selected with HAT medium. Antibodies to taxol were screened by indirect ELISA. The hybridoma cells that secrete high affinity monoclonal antibodies against taxol were cloned by a limiting dilution method. Meanwhile, the conditions of indirect ELISA for determination of antibody against taxol were optimized. Results; Three hybridoma cell lines secreting the antibody, taxol-Al, taxol-A2 and taxol-A3 , were obtained and the antibodies had been purified from ascites using caprylic acid/ammonium sulfate precipitation (CA-AS). Subclasses of the purified antibodies belonged to IgG2b. The titer of antibody from the hybridoma celltaxol-A3-induced ascites was 1: 128 000. The optimal detection conditions were 15 (xg-mL"1 of antigen in coating buffer, at 4 t , overnight, 1 % BSA blocking for 1 h, PBS as dilution solution, 30 min of MeAb reaction time, 1 h of enzyme-labeled antibody reaction time, and termination of reaction with 10% H2SO4. Conclusion; The monoclonal antibodies against taxol have been prepared, and the indirect ELISA method for rapidly detecting taxol is successfully established. This method provided an important basis for the detection of taxol produced by microbe fermentation in the future.%目的:制备抗紫杉醇单克隆抗体并建立间接ELISA检测方法.方法:用琥珀酸酐法将紫杉醇(taxol)与牛血清白蛋白(BSA)偶联制成全抗原taxol-BSA,采用紫外波长扫描法和薄层层析

  5. Flexural Behavior of Aluminum Honeycomb Core Sandwich Structure

    Science.gov (United States)

    Matta, Vidyasagar; Kumar, J. Suresh; Venkataraviteja, Duddu; Reddy, Guggulla Bharath Kumar

    2017-05-01

    and has more strength. By the power press used as forming method we fabricate the honey comb core and stacking the sheets with adhesive as epoxy resin or laser beam welding and sandwich structure will form with two face sheets. Then the specimen is taken to be tested to know the flexural behaviour by the flexural test as 3 point and 4 pont bend test. After testing of two different tests then we get the force vs displacement curve by this we can know the maximum force and by loading configurations and its displacement or deflection then we can calculate flexural stiffness and core shear modulus by the variation of three parameters. Our ultimate aim is to achieve maximum strength by minimum weight.

  6. Validation of ELISA for the detection of African horse sickness virus antigens and antibodies.

    Science.gov (United States)

    Rubio, C; Cubillo, M A; Hooghuis, H; Sanchez-Vizcaino, J M; Diaz-Laviada, M; Plateau, E; Zientara, S; Crucière, C; Hamblin, C

    1998-01-01

    The mortality rate in susceptible populations of horses during an epizootic of African horse sickness (AHS) may be in excess of 90%. Rapid and reliable assays are therefore essential for the confirmation of clinical diagnoses and to enable control strategies to be implemented without undue delay. One of the major objectives of a recent European Union funded project was the validation of newly developed diagnostic assays which are rapid, sensitive, highly reproducible and inexpensive, for the detection of African horse sickness virus (AHSV) antigens and antibodies. The Laboratorio de Sanidad y Produccion Animal (LSPA) in Algete, Spain was charged with the responsibility of co-ordinating and supplying samples of viruses and antisera to the participating laboratories in Spain, France and the United Kingdom. The panels comprised 76 antigen samples for assay by indirect sandwich ELISAs and 53 serum samples for antibody detection by either indirect or competitive ELISAs. Results generated by ELISA for each laboratory were analysed in LSPA in terms of their relative sensitivities and specificities. There was a good agreement between the ELISAs used for either antigen or antibody detection. The participating groups agreed that any field sample giving a doubtful result would always be retested by ELISA and an alternative assay.

  7. ELISA testing for common food antigens in four dry dog foods used in dietary elimination trials.

    Science.gov (United States)

    Raditic, D M; Remillard, R L; Tater, K C

    2011-02-01

    This study evaluated four over the counter venison dry dog foods available from one on-line retail vendor for potential contamination with common known food allergens: soy, poultry or beef. An amplified, double sandwich type enzyme linked immunosorbent assay (ELISA) test of soy, poultry and beef proteins were performed by an independent accredited food laboratory. The ELISA test for poultry protein was found to be unreliable when testing in dry dog foods because false negatives occurred. ELISA testing of control diets for both soy and beef proteins performed as expected and could be useful in antigen testing in dry dog foods. Three of the four over the counter (OTC) venison canine dry foods with no soy products named in the ingredient list were ELISA positive for soy; additionally one OTC diet tested positive for beef protein with no beef products listed as an ingredient list. One OTC venison diet was not found to be positive for soy, poultry or beef proteins. However, none of the four OTC venison diets could be considered suitable for a diagnostic elimination trial as they all contained common pet food proteins, some of which were readily identifiable on the label and some that were only detected by ELISA. Therefore, if the four OTC venison products selected in this study are representative of OTC products in general, then the use of OTC venison dry dog foods should not be used during elimination trials in suspected food allergy patients. © 2010 Blackwell Verlag GmbH.

  8. PREPARATION OF MONOCLONAL ANTIBODY AND DEVELOPMENT OF AN INDIRECT COMPETITIVE ELISA METHOD FOR GENTAMICIN%庆大霉素单克隆抗体的研制及ELISA分析方法的建立

    Institute of Scientific and Technical Information of China (English)

    金仁耀; 吴建祥

    2013-01-01

    One hybridoma cell line (6H8) secreting monoclonal antibody (McAb) against gentamicin was produced by fusing mouse myeloma cells ( SP2/0) with spleen cells from BALB/C which immunized by the artificial gentamicin antigen conjugated with bovine serum albumin ( BSA). Isotype and subclass of the monoclonal antibody secreted from the hybridoma cell line (6H8) was classified as IgGl. The light chain of the McAb was identified to be Κ chain. The McAb obtained could specifically react with gentamicin and its titre of ascitic fluid detected by indirect ELISA was up to 1 × 10-7. The result of specificity analysis indicated that the McAb had no cross-reactivity with analogues of gentamicin. Based on the producted McAb, an indirect competitive ELISA was established, and its linear regression equation was y = 16. 1221n (x) -2. 0143 (R2 =0. 9934). Inhibition rate analysis showed that IC50 and IC20 values were 25. 2 μg·L-1 and 3. 9 μg·L-1 gentamicin in PBS buffer, respectively. The mean recovery of gentamicin spiked in milk was from 91% to 110%. The producted anti-gentamicin McAb and established competitive ELISA method could lay the foundation for rapid detection of gentamicin residue.%用与牛血清蛋白(BSA)交联的庆大霉素人工抗原(GM-BSA)免疫的BALB/C鼠脾细胞与SP2/0鼠骨髓瘤细胞融合,经筛选、克隆,得到1株能稳定分泌抗庆大霉素单抗的杂交瘤细胞株(6H8),经鉴定6H8的抗体类型及亚类均为IgG1,其轻链为κ链.制备单克隆抗体腹水,腹水的间接ELISA效价在1×10-7以上.该单克隆抗体与庆大霉素结构类似物均无交叉反应,具有高度特异性.以制备的单抗建立间接竞争ELISA方法,其线性回归方程为y=16.122ln(x)-2.0143(R2=0.9934),抑制中浓度IC50为25.2μg·L-1,最低检测限IC20为3.9μg·L-1.竞争ELISA方法检测鲜奶中的庆大霉素平均回收率在92%~110%之间.抗庆大霉素单抗的制备和竞争ELISA方法的建立为庆大霉素快速检测奠定了基础.

  9. Combined compressive and shear buckling analysis of hypersonic aircraft sandwich panels

    Science.gov (United States)

    Ko, William L.; Jackson, Raymond H.

    1992-01-01

    The combined-load (compression and shear) buckling equations were established for orthotropic sandwich panels by using the Rayleigh-Ritz method to minimize the panel total potential energy. The resulting combined-load buckling equations were used to generate buckling interaction curves for super-plastically-formed/diffusion-bonded titanium truss-core sandwich panels and titanium honeycomb-core sandwich panels having the same specific weight. The relative combined-load buckling strengths of these two types of sandwich panels are compared with consideration of their sandwich orientations. For square and nearly square panels of both types, the combined load always induces symmetric buckling. As the panel aspect ratios increase, antisymmetric buckling will show up when the loading is shear-dominated combined loading. The square panel (either type) has the highest combined buckling strength, but the combined load buckling strength drops sharply as the panel aspect ratio increases. For square panels, the truss-core sandwich panel has higher compression-dominated load buckling strength. However, for shear dominated loading, the square honeycomb-core sandwich panel has higher shear-dominated combined load buckling strength.

  10. Combined compressive and shear buckling analysis of hypersonic aircraft structural sandwich panels

    Science.gov (United States)

    Ko, William L.; Jackson, Raymond H.

    1991-01-01

    The combined-load (compression and shear) buckling equations were established for orthotropic sandwich panels by using the Rayleigh-Ritz method to minimize the panel total potential energy. The resulting combined-load buckling equations were used to generate buckling interaction curves for super-plastically-formed/diffusion-bonded titanium truss-core sandwich panels and titanium honeycomb-core sandwich panels having the same specific weight. The relative combined-load buckling strengths of these two types of sandwich panels are compared with consideration of their sandwich orientations. For square and nearly square panels of both types, the combined load always induces symmetric buckling. As the panel aspect ratios increase, antisymmetric buckling will show up when the loading is shear-dominated combined loading. The square panel (either type) has the highest combined buckling strength, but the combined load buckling strength drops sharply as the panel aspect ratio increases. For square panels, the truss-core sandwich panel has higher compression-dominated combined load buckling strength. However, for shear dominated loading, the square honeycomb-core sandwich panel has higher shear-dominated combined load buckling strength.

  11. [Sensitivity and specificity of the ELISA Kit for the detection of antidobies to Junin virus].

    Science.gov (United States)

    Pirozhkov, A P; Timofeev, M A; Borisevich, I V; Syromiatnikova, S I; Shatokhina, I V; Pantyukhov, V B; Kovalchuk, A V; Borisevich, S V

    2015-01-01

    The goal of this work was to describe methodological approaches to determination of sensitivity and specificity of the enzyme-linked immunosorbent assay kit (ELISA Kit) for detection of the specific anti-Junin virus (JV) antibody. Comparison of ELISA to plaque reduction neutralization test (PRNT) showed direct relationship between antibody titers in the samples of serum of immunized animals, determined by either PRNT or ELISA methods. The obtained results provided an opportunity to form the panels of positive and negative serum samples to determine the sensitivity and specificity of the ELISA Kit. Sensitivity of the ELISA Kit was at least 98% when studying the samples of serum of immunized guinea pigs and rabbits (determined as positive in PRNT). The sensitivity of the ELISA Kit was at least 68% when studying the samples determined by PNRT as uncertain positive. The specificity was 98%. The specificity of the ELISA Kit was 98%.

  12. Monoclonal antibody-based serological methods for detection of Cucumber green mottle mosaic virus

    Directory of Open Access Journals (Sweden)

    Qian Yajuan

    2011-05-01

    Full Text Available Abstract Background Cucumber green mottle mosaic virus (CGMMV, a member of the genus Tobamovirus, can be transmitted by seeds and infects many cucurbit species, causing serious yield losses in cucumber and watermelon plants. In this paper, five serological methods including antigen-coated plate enzyme-linked immunosorbent assay (ACP-ELISA, triple antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA, Dot-immunobinding assay (DBIA, direct tissue blot immunoassay (DTBIA and immunocapture reverse transcriptase polymerase chain reaction (IC-RT-PCR were described for detection and diagnosis of CGMMV. Results Using the purified CGMMV particles as immunogens, six murine monoclonal antibodies (MAbs were produced. Five serological methods were established using the MAb 4H1 and detection sensitivity was compared using purified preparations and infected-plant tissue extracts. The detection sensitivity of ACP-ELISA was 0.16 ng of purified CGMMV, whereas TAS-ELISA was more sensitive than ACP-ELISA with a minimum detection of 0.04 ng of purified CGMMV. The sensitivities of TAS-ELISA and DBIA were similar for detecting CGMMV in infected-plant tissue extracts, and were four times higher than ACP-ELISA. The IC-RT-PCR was the most sensitive method, which could detect as little as 0.1 pg of purified virus. The detection sensitivity of IC-RT-PCR for CGMMV-infected plant tissues was about 400 times higher than that of TAS-ELISA and DBIA. Conclusions The established ACP-ELISA, TAS-ELISA, DBIA and DTBIA are suitable for routine CGMMV detection of large-scale samples in the field survey, while IC-RT-PCR is more sensitive and suitable for acquiring information about the viral genome.

  13. Analysis of Grid-Scored Sandwich Structures of Different Curvatures and Grid Sizes For Wind Turbine Blades

    DEFF Research Database (Denmark)

    Laustsen, Steffen; Thomsen, Ole Thybo; Lund, Erik;

    2012-01-01

    The stress and strain field developed locally in-situ the core of grid-scored sandwich structures in wind turbine blades is investigated. Due to the many singularities occurring from the “tri-material corners”, a full 3D analysis of the sandwich structure in terms of the Finite Element Method...

  14. Theoretical Calculation and Analysis of Slip and Deformation for Concrete Sandwich Panels

    Institute of Scientific and Technical Information of China (English)

    LI Yanbo; ZHANG Shaohua; XIA Baoyang

    2007-01-01

    Slip and deformation of concrete sandwich panels under uniformly distributed loads is concerned. The effect of slip on the deformation of concrete sandwich panels are studied, and the analytical expressions of slip and deformation for concrete sandwich panels is obtained. These formulae can describe the slip distribution and account for its effect on deformation. In order to restrict the bound of formula, the formula of crack moment is obtained. The results of theoretical calculation are compared with those of tests and finite element methods. The comparison shows that the results of theoretical calculation are in accord with those of tests and finite element methods. So the theoretical calculation can be used to calculate slip and deformation of concrete sandwich panels in practical projects.

  15. Comparative study of circulating immune complexes quantity detection by three assays--CIF-ELISA, C1q-ELISA and anti-C3 ELISA.

    Science.gov (United States)

    Stanilova, S A; Slavov, E S

    2001-07-01

    The assessment of the soluble immune complexes (IC) in human sera is traditionally performed by the C1q binding assay. In the present study, a novel method for the quantity of immune complexes was reported. The methodology was based on measuring their deposition on solid-phase C3 binding glycoprotein (CIF), using an enzyme-linked immunosorbent assay. We also used ELISA that employed anti-C3 antibodies to determined the quantity of immune complexes. The three assays were evaluated for their performance characteristics on the same specially prepared samples: 55 normal sera, 99 sera from RA, 88 sera from SLE, and 27 sera from PSS. The results were compared by reference to a common standard-heat aggregated IgG that possesses many activities of immune complexes. Three of the tests used displayed almost the same specificity (over 95%), while their relative sensitivity varied depending on the disease sera tested. The sensitivity of the assays used was recorded highest for C1q ELISA-28.97% of positive sera, followed by CIF-ELISA-19.63% and lowest for anti-C3 ELISA-17.29%. A well-expressed correlation was found between CIF-ELISA and anti-C3 ELISA data (r=0.42), and a week correlation was noted when comparing CIF-ELISA and C1q ELISA IC levels detected (r=0.28). When the correlation coefficients were calculated individually for each disease category, they were clearly different, and that reflected indirectly in different sensitivities of the test for various disease categories. We also found that the results from the simultaneous performance of the tests demonstrated low percentage positive results when three or two assays were used. This is most probably due to the different assay abilities to detect IC with different sizes and composition, which shows that a small part of IC in the tested sera can be detected simultaneously by more than one assay. On the basis of the results obtained, we concluded that optimal screening for IC could be achieved by parallel application of

  16. Prospective Comparison of the Diagnostic Potential of Real-Time PCR, Double-Sandwich Enzyme-Linked Immunosorbent Assay for Galactomannan, and a (1→3)-β-d-Glucan Test in Weekly Screening for Invasive Aspergillosis in Patients with Hematological Disorders

    Science.gov (United States)

    Kawazu, Masahito; Kanda, Yoshinobu; Nannya, Yasuhito; Aoki, Katsunori; Kurokawa, Mineo; Chiba, Shigeru; Motokura, Toru; Hirai, Hisamaru; Ogawa, Seishi

    2004-01-01

    The establishment of an optimal noninvasive method for diagnosing invasive aspergillosis (IA) is needed to improve the management of this life-threatening infection in patients with hematological disorders, and a number of noninvasive tests for IA that target different fungal components, including galactomannan, (1→3)-β-d-glucan (BDG), and Aspergillus DNA, have been developed. In this study, we prospectively evaluated the diagnostic potential of three noninvasive tests for IA that were used in a weekly screening strategy: the double-sandwich enzyme-linked immunosorbent assay (ELISA) for galactomannan (Platelia Aspergillus), a real-time PCR assay for Aspergillus DNA (GeniQ-Asper), and an assay for BDG (β-glucan Wako). We analyzed 149 consecutive treatment episodes in 96 patients with hematological disorders who were at high risk for IA and diagnosed 9 proven IA cases, 2 probable IA cases, and 13 possible invasive fugal infections. In a receiver-operating characteristic (ROC) analysis, the area under the ROC curve was greatest for ELISA, using two consecutive positive results (0.97; P = 0.036 for ELISA versus PCR, P = 0.055 for ELISA versus BDG). Based on the ROC curve, the cutoff for the ELISA could be reduced to an optical density index (O.D.I.) of 0.6. With the use of this cutoff for ELISA and cutoffs for PCR and BDG that give a comparable level of specificity, the sensitivity/specificity/positive predictive value/negative predictive value of the ELISA and the PCR and BDG tests were 1.00/0.93/0.55/1.00, 0.55/0.93/0.40/0.96, and 0.55/0.93/0.40/0.96, respectively. In conclusion, among these weekly screening tests for IA, the double-sandwich ELISA test was the most sensitive at predicting the diagnosis of IA in high-risk patients with hematological disorders, using a reduced cutoff of 0.6 O.D.I. PMID:15184460

  17. Prospective comparison of the diagnostic potential of real-time PCR, double-sandwich enzyme-linked immunosorbent assay for galactomannan, and a (1-->3)-beta-D-glucan test in weekly screening for invasive aspergillosis in patients with hematological disorders.

    Science.gov (United States)

    Kawazu, Masahito; Kanda, Yoshinobu; Nannya, Yasuhito; Aoki, Katsunori; Kurokawa, Mineo; Chiba, Shigeru; Motokura, Toru; Hirai, Hisamaru; Ogawa, Seishi

    2004-06-01

    The establishment of an optimal noninvasive method for diagnosing invasive aspergillosis (IA) is needed to improve the management of this life-threatening infection in patients with hematological disorders, and a number of noninvasive tests for IA that target different fungal components, including galactomannan, (1-->3)-beta-d-glucan (BDG), and Aspergillus DNA, have been developed. In this study, we prospectively evaluated the diagnostic potential of three noninvasive tests for IA that were used in a weekly screening strategy: the double-sandwich enzyme-linked immunosorbent assay (ELISA) for galactomannan (Platelia Aspergillus), a real-time PCR assay for Aspergillus DNA (GeniQ-Asper), and an assay for BDG (beta-glucan Wako). We analyzed 149 consecutive treatment episodes in 96 patients with hematological disorders who were at high risk for IA and diagnosed 9 proven IA cases, 2 probable IA cases, and 13 possible invasive fugal infections. In a receiver-operating characteristic (ROC) analysis, the area under the ROC curve was greatest for ELISA, using two consecutive positive results (0.97; P = 0.036 for ELISA versus PCR, P = 0.055 for ELISA versus BDG). Based on the ROC curve, the cutoff for the ELISA could be reduced to an optical density index (O.D.I.) of 0.6. With the use of this cutoff for ELISA and cutoffs for PCR and BDG that give a comparable level of specificity, the sensitivity/specificity/positive predictive value/negative predictive value of the ELISA and the PCR and BDG tests were 1.00/0.93/0.55/1.00, 0.55/0.93/0.40/0.96, and 0.55/0.93/0.40/0.96, respectively. In conclusion, among these weekly screening tests for IA, the double-sandwich ELISA test was the most sensitive at predicting the diagnosis of IA in high-risk patients with hematological disorders, using a reduced cutoff of 0.6 O.D.I.

  18. New "sandwich" structures conformed from three dimensional

    Directory of Open Access Journals (Sweden)

    Alba, Juan J.

    1996-03-01

    Full Text Available Poor interlaminar properties as well as poor-skin-to-core adhesion properties are very often the common existing problems we find when designing with "sandwich" structures. A new type of 3D-fabric "sandwich" structure is being developed in order to avoid these problems. Although the manufacturing process is very simple, a very complex "sandwich" structure is obtained as a result of the complexity of the 3D-fabric used. This 3D-fabric is a 3D woven glass fabric produced on velvet weaving machines with glass yarns. It is an integrally woven "sandwich" laminate for all kinds of composite products. The strength of the vertical fibers makes, that also after impregnation with a resin matrix, the "sandwich" structure is maintained. The result is a laminate with high strength and stiffness and low weight. On each side of this "sandwich" laminate additional reinforcement materials can be laminated and a synthetic foam can be injected in the hollow structure. This will allow to establish the mechanical properties of a finished product.

    Las pobres propiedades, tanto interlaminares como de adhesión entre piel y núcleo, constituyen uno de los grandes problemas cuando se diseñan estructuras utilizando paneles tipo "sandwich". Un nuevo tipo de panel "sandwich", configurado a partir de tejidos tridimensionales, está siendo desarrollado en la actualidad con el objetivo de eliminar esos problemas. Aunque el proceso de fabricación es muy simple, el panel "sandwich" obtenido es de estructura compleja, como resultado de la complejidad del tejido tridimensional utilizado. Este tejido tridimensional (3D es un tejido de fibra de vidrio producido en máquinas de tejer especializadas. La resistencia de las fibras verticales hace que, después de la impregnación con una resina, se mantenga la configuración tipo "sandwich". El resultado es un laminado de alta resistencia, gran rigidez y bajo peso. Sobre cada uno de los lados del panel "sandwich" se pueden

  19. New "sandwich" structures conformed from three dimensional

    OpenAIRE

    1996-01-01

    Poor interlaminar properties as well as poor-skin-to-core adhesion properties are very often the common existing problems we find when designing with "sandwich" structures. A new type of 3D-fabric "sandwich" structure is being developed in order to avoid these problems. Although the manufacturing process is very simple, a very complex "sandwich" structure is obtained as a result of the complexity of the 3D-fabric used. This 3D-fabric is a 3D woven glass fabric produced on velvet weaving machi...

  20. HCV胶体金法与ELISA法在输血前检测丙型肝炎抗体中的效果比较%Effect comparison of HCV colloidal gold method and ELISA method in detection hepatitis c antibody before blood transfusion

    Institute of Scientific and Technical Information of China (English)

    钟佩怡; 邓常春; 陈丽萍; 易素芬; 陈瑞芬

    2014-01-01

    目的:HCV胶体金法与ELISA法用于输血前检查丙型肝炎抗体检测的应用效果比较。方法:分别采用ELISA法和胶体金法对我院576名受血者的血清样本进行HCV-抗体检测,对两种检测方法的检测效果及检测用时及成本预算进行综合比较。结果:ELISA法阳性检测率高于胶体金法,但比较差异无统计学意义(P>0.05);经胶体金法和ELISA检测为阳性的血清样本再使用RT-PCR 荧光定量法检测,胶体金法的检测符合率达到100%,显著高于ELISA法(P0.05) difference comparison;The colloidal gold method and ELISA test for positive serum samples using the fluorescent quantitative rt-pcr method to detect, colloidal gold method detection coincidence rate reached 100%, significantly higher than that of ELISA method (P < 0.05); Colloidal gold test required for testing time is short and the cost is lower. Conclusions:compared with ELISA method, colloidal gold method is used to detect lower false positive rate of hepatitis c virus (HCV), operation more convenient, cost is low, worthy of clinical popularization and application.

  1. 纤维蛋白胶抗体间接ELISA检测方法的建立及初步应用%Development and preliminary application of indirect ELISA method for fibrin glue antibody

    Institute of Scientific and Technical Information of China (English)

    蔡阳; 任真; 何学军; 吴华; 乔红群

    2012-01-01

    Objective To develop and preliminarily apply an indirect ELISA method for fibrin glue antibody. Methods An indirect ELISA method for fibrin glue antibody was developed using fibrin glue as coating antigen, of which the reaction condition was optimized, and precision and specificity were verfied. The serum antibody of rabbits immunized with fibrin glue was determined by the developed method. Results The reaction condition for the developed indirect ELJSA method was optimized as follows: microtiter plate was irradiated vertically with UV light for 20 min, then coated with fibrin glue at a concentration of 10 |xg/ml, diluted with 0. 05 mol/L bicarbonate solution (pH 9. 6) as coating buffer solution, at 4℃ overnight; the PBS containing 10% fetal bovine serum was served as blocking solution, while 0. 1% PBST as antibody diluent; the optimal dilutions of polyclonal antibody against fibrin glue and HRP-labeled goat anti-rabbit IgG were 1 : 2 500 and 1 : 4 000 respectively. The optimal temperature and time for reaction were 37℃ and 1 h, while those for substrate coloration were 37℃ and 15 min, respectively. The sulfuric acid at a concentration of 2 mol / L was served as stop solution. The method showed high precision and specificity. The antibodies in sera of rabbits 1,2,4 and 6 weeks after immunization were determined, and the result showed that the serum antibody level increased significantly 2 weeks while started to decrease 6 weeks after immunization. Conclusion An indirect ELISA method for fibrin glue antibody was successfully developed, which might be used for the determination of fibrin glue antidoy in immune sera of rabbits.%目的 建立纤维蛋白胶抗体间接ELISA检测方法,并进行初步应用.方法 采用纤维蛋白胶作为包被抗原,建立检测纤维蛋白胶抗体的间接ELISA法,优化反应条件,并对方法的精密性及特异性进行验证.采用建立的间接ELISA法检测纤维蛋白胶免疫兔血清抗体.结果 间接ELISA法

  2. Non-Uniform Compressive Strength of Debonded Sandwich Panels

    DEFF Research Database (Denmark)

    Berggreen, Carl Christian; Simonsen, Bo Cerup

    2005-01-01

    This article describes the development, validation and application of a FEM based numerical model for prediction of residual strength of damaged sandwich panels. The core of the theoretical method is a newly developed procedure for prediction of the propagation of a face-core debond. As demonstra......This article describes the development, validation and application of a FEM based numerical model for prediction of residual strength of damaged sandwich panels. The core of the theoretical method is a newly developed procedure for prediction of the propagation of a face-core debond.......(2005)., shows that the model is indeed able to predict the failure modes and the residual strength of damaged panels with accuracy sufficient for practical applications. This opens up for a number of important engineering applications, for example risk-based inspection and repair schemes....

  3. Seroprevalence of antibodies to influenza A/H1N1/2009 among transmission risk groups after the second wave in Mexico, by a virus-free ELISA method

    Science.gov (United States)

    Elizondo-Montemayor, Leticia; Alvarez, Mario M.; Hernández-Torre, Martín; Ugalde-Casas, Patricia A.; Lam-Franco, Lorena; Bustamante-Careaga, Humberto; Castilleja-Leal, Fernando; Contreras-Castillo, Julio; Moreno-Sánchez, Héctor; Tamargo-Barrera, Daniela; López-Pacheco, Felipe; Freiden, Pamela J.; Schultz-Cherry, Stacey

    2014-01-01

    Summary Objective No serological studies have been performed in Mexico to assess the seroprevalence of influenza A/H1N1/2009 in groups of people according to the potential risk of transmission. The aim of this study was to determine the seroprevalence of antibodies against influenza A/H1N1/2009 in subjects in Mexico grouped by risk of transmission. Methods Two thousand two hundred and twenty-two subjects were categorized into one of five occupation groups according to the potential risk of transmission: (1) students, (2) teachers, (3) healthcare workers, (4) institutional home residents aged >60 years, and (5) general population. Seroprevalence by potential transmission group and by age grouped into decades was determined by a virus-free ELISA method based on the recombinant receptor-binding domain of the hemagglutinin of influenza A/H1N1/2009 virus as antigen (85% sensitivity; 95% specificity). The Wilson score, Chi-square test, and logistic regression models were used for the statistical analyses. Results Seroprevalence for students was 47.3%, for teachers was 33.9%, for older adults was 36.5%, and for the general population was 33.0%, however it was only 24.6% for healthcare workers (p = 0.011). Of the students, 56.6% of those at middle school, 56.4% of those at high school, 52.7% of those at elementary school, and 31.1% of college students showed positive antibodies (p < 0.001). Seroprevalence was 44.6% for college teachers, 31.6% for middle school teachers, and 29.8% for elementary school teachers, but was only 20.3% for high school teachers (p = 0.002). Conclusions The student group was the group most affected by influenza A/H1N1/2009, while the healthcare worker group showed the lowest prevalence. Students represent a key target for preventive measures. PMID:21855383

  4. A Quick Pretreatment Method for Determination of Chloramphenicol Residue in Royal Jelly by ELISA%ELISA检测蜂王浆氯霉素残留的快速前处理方法

    Institute of Scientific and Technical Information of China (English)

    李璟; 陈世界; 李凛; 童晋; 聂春梅; 陈洋

    2012-01-01

    We used the improved method (0. 1 mol/L Tris-TBE) to deal with royal jelly samples, then extracted chloramphenicol (CAP) with ethyl acetate. The whole processing only needs 25 min. Compare with the original method from the ELISA kit the OD450 values of the improved method were lower than those of the kit method. We added standard CAP solution to the samples treated by the improved method to verify the sensitivity of it. The OD450 value changed which were obviously different from samples without adding CAP(F<0. 05). It was indicated that there was no interfering agent in the process of pretreatment and coloration. The average RSD of enzyme-marked board is 1. 29% . wich is accord with the request of international determination of chloramphenicol residue. Our assay provides a quick pretreatment method to detect the CAP in royal jelly. It is useful in the high-throughput detection for the inspection and quarantine departments.%为获得一种能快速检测蜂王浆中氯霉素残留的前处理方法,采用改良的0.1 mol/L Tris-TBE(pH=8.0)溶液处理蜂王浆,结合乙酸乙酯提取氯霉素(CAP),处理时间约25 min.ELISA检测样品的OD450值明显低于试剂盒法,通过添加标准CAP溶液对改良法进行验证.结果表明,改良法提取的样品对不同CAP质量浓度表现出明显的 OD450值变化,与添加前比较差异显著(P<0.05),排除处理过程存在干扰显色的因素.改良法检测相同样品的平均相对标准偏差为1.29%,完全符合目前国际上氯霉素残留检测的要求.

  5. An asymptotically exact theory of smart sandwich shells

    CERN Document Server

    Le, Khanh Chau

    2016-01-01

    An asymptotically exact two-dimensional theory of elastic-piezoceramic sandwich shells is derived by the variational-asymptotic method. The error estimation of the constructed theory is given in the energetic norm. As an application, analytical solution to the problem of forced vibration of a circular elastic plate partially covered by two piezoceramic patches with thickness polarization excited by a harmonic voltage is found.

  6. Standardization of allergen products: 3. Validation of candidate European Pharmacopoeia standard methods for quantification of major birch allergen Bet v 1.

    Science.gov (United States)

    Kaul, S; Zimmer, J; Dehus, O; Costanzo, A; Daas, A; Buchheit, K H; Asturias, J A; Barber, D; Carnés, J; Chapman, M; Dayan-Kenigsberg, J; Döring, S; Führer, F; Hanschmann, K M; Holzhauser, T; Ledesma, A; Moingeon, P; Nony, E; Pini, C; Plunkett, G; Reese, G; Sandberg, E; Sander, I; Strecker, D; Valerio, C; van Ree, R; Vieths, S

    2016-10-01

    The BSP090 project aims at establishing European Pharmacopoeia Reference Substances in combination with the corresponding ELISA methods for the quantification of major allergens in allergen products. Two sandwich ELISAs proved suitable for quantification of Bet v 1, the major birch pollen allergen, in preceding phases of BSP090. Two Bet v 1-specific ELISA systems were compared with respect to accuracy and precision in a ring trial including 13 laboratories. Model samples containing recombinant rBet v 1.0101 as well as native birch pollen extracts were measured independently at least three times in each facility. The assessment was completed with a comparative quantification of Bet v 1 in 30 marketed birch allergen products in one laboratory, simulating the future use as reference method. In the collaborative study, both candidate ELISAs confirmed their suitability to quantify recombinant and native Bet v 1. ELISA-A showed higher precision and lower interlaboratory variability, yet ELISA-B exhibited slightly higher accuracy. Subsequent parallel measurement of Bet v 1 in a panel of 'real-life' birch allergen products indicated better repeatability of ELISA-B. Both systems detected substantial differences in Bet v 1 content between allergen products, but the effect was more pronounced using ELISA-B due to persistently higher values compared to ELISA-A. In the collaborative study, no deciding differences were observed between the two candidate ELISAs. Further comparison under conditions simulating the intended use combined with the criterion of long-term availability enabled the selection of one Bet v 1-specific ELISA for proposal as European Pharmacopoeia standard method. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  7. Buckling driven debonding in sandwich columns

    DEFF Research Database (Denmark)

    Østergaard, Rasmus Christian

    2008-01-01

    A compression loaded sandwich column that contains a debond is analyzed using a geometrically non-linear finite element model. The model includes a cohesive zone along one face sheet/core interface whereby the debond can extend by interface crack growth. Two geometrical imperfections are introduced......; a global imperfection of the sandwich column axis and a local imperfection of the debonded face sheet axis. The model predicts the sandwich column to be very sensitive to the initial debond length and the local face sheet imperfection. The study shows that the sensitivity to the face sheet imperfection...... results from two mechanisms: (a) interaction of local debond buckling and global buckling and (b) the development of a damaged zone at the debond crack tip. Based on the pronounced imperfection sensitivity, the author predicts that an experimental measurement of the strength of sandwich structures may...

  8. Local slamming impact of sandwich composite hulls

    National Research Council Canada - National Science Library

    Qin, Z; Batra, R.C

    2009-01-01

    We develop a hydroelastic model based on a {3,2}-order sandwich composite panel theory and Wagner's water impact theory for investigating the fluid-structure interaction during the slamming process...

  9. The application of ELISA method on serum antibody of hepatitis C virus in different groups%ELISA法对不同人群血清丙型肝炎病毒抗体检测应用

    Institute of Scientific and Technical Information of China (English)

    朱德春

    2014-01-01

    目的:应用酶联免疫吸附法(Enzyme-Linked ImmunoSorbent Assay, ELISA)检测不同人群丙型肝炎感染情况,为丙型肝炎防治工作提供依据。方法:采用分层抽样方法抽取我市12305例对象,应用ELISA法检测不同人群血清丙型肝炎病毒(Hepatitis C virus,HCV)抗体。结果:本次调查共有393例调查对象呈H C V抗体阳性,人群H C V平均感染率为3.19%,与全国平均水平(3.22%)比较无统计学差异(P<0.05)。一般人群、普通孕妇、医护人员、不良接触史者、乙肝病人和血液透析病人抗-H C V阳性率分别为0.60%,0.78%,3.17%,33.00%,17.00%和55.24%。血液透析病人、不良接触史者、乙肝病人和医护人员抗-H C V阳性率显著高于一般人群(P<0.05)。普通孕妇与一般人群抗-HCV阳性率比较无统计学差异(P>0.05)。不同年龄医护人员H C V抗体阳性率比较有统计学差异(P<0.05)。男性H C V抗体阳性率显著高于女性(P<0.05),有输血史的血液透析患者HCV抗体阳性率显著高于无输血史患者(P<0.05)。结论:输血是H C V感染的主要传播途径,应对高危人群开展健康干预,同时医护人员的防护也应得到重视。%Objective:Used enzyme linked immunosorbent assay (ELISA) to detect the infection of hepatitis C that providing basis for the prevention and control of hepatitis C.Methods:Selected 12305 cases of different populations by stratified sampling method, used ELISA to detect serum hepatitis C virus antibody.Results:393 subjects were HCV antibody positive, HCV population, the average infection rate was 3.19%, there was no signiifcant difference with the national average level (3.22%)(P0.05). The positive rate of HCV antibody in different age medical staff were statistically signiifcant difference (P<0.05). The positive rate of HCV antibody was male female(P<0.05), the positive rate of HCV antibody in hemodialysis

  10. Novel sandwich structure adsorptive membranes for removal of 4-nitrotoluene from water

    Energy Technology Data Exchange (ETDEWEB)

    Guo, Yuexin [College of Chemistry, Beijing Normal University, Beijing 100875 (China); School of Pharmacy, North China University of Science and Technology, Tangshan 063000 (China); Jia, Zhiqian, E-mail: zhqjia@bnu.edu.cn [College of Chemistry, Beijing Normal University, Beijing 100875 (China)

    2016-11-05

    Highlights: • Novel sandwich PES-SPES/PS-PDVB/PTFE adsorptive membranes were prepared. • The removal efficiency for 4-nitrotoluene is greater than 95% after five recycles. • The membrane showed higher adsorption capacity than that of mixed matrix membrane. - Abstract: Novel sandwich PES-SPES/PS-PDVB/PTFE adsorptive membranes were prepared by a filtration/immersion precipitation method and employed for the removal of 4-nitrotoluene from water. The static adsorption thermodynamics, kinetics, dynamic adsorption/desorption and membrane reusability were investigated. The results showed that the Freundlich model describes the adsorption isotherm satisfactorily. With increased PS-PDVB content, the maximum static adsorption capacity, partition coefficient, apparent adsorption rate constant, and dynamic adsorption capacity all significantly increased. The sandwich membranes showed much higher removal efficiency and adsorption capacity than those of mixed matrix membranes. With respect to dynamics adsorption/desorption, the sandwich membranes exhibited excellent reusability, with a removal efficiency greater than 95% even after five recycles.

  11. Vibroacoustic optimization of anti-tetrachiral and auxetic hexagonal sandwich panels with gradient geometry

    Science.gov (United States)

    Ranjbar, Mostafa; Boldrin, Luca; Scarpa, Fabrizio; Neild, Simon; Patsias, Sophoclis

    2016-05-01

    The work describes the vibroacoustic behavior of anti-tetrachiral and auxetic hexagonal gradient sandwich panels using homogenized finite element models to determine the mechanical properties of the auxetic structures, the natural frequencies and radiated sound power level of sandwich panels made by the auxetic cores. The mechanical properties and the vibroacoustic behavior of auxetic hexagonal sandwich panels are investigated as a benchmark. The radiated sound power level of the structure over the frequency range of 0-1000 Hz is minimized by modifying the core geometry of the gradient auxetic sandwich panels. Several excitation cases are considered. First-order and random optimization methods are used for the minimization of radiated sound power level of the structures. The results of this study present significant insights into the design of auxetic structures with respect to their vibroacoustical properties.

  12. Stress Distribution on Sandwich Structure with Triangular Grid Cores Suffered from Bending Load

    Directory of Open Access Journals (Sweden)

    Cui Xu

    2015-01-01

    Full Text Available Triangular grid reinforced by carbon fiber/epoxy (CF/EP was designed and manufactured. The sandwich structure was prepared by gluing the core and composite skins. The mechanical properties of the sandwich structure were investigated by the finite element analysis (FEA and three-point bending methods. The calculated bending stiffness and core shear stress were compared to the characteristics of a honeycomb sandwich structure. The results indicated that the triangular core ultimately failed under a bending load of 11000 N; the principal stress concentration was located at the loading region; and the cracks occurred on the interface top skin and triangular core. In addition, the ultimate stress bearing of the sandwich structure was 8828 N. The experimental results showed that the carbon fiber reinforced triangular grid was much stiffer and stronger than the honeycomb structure.

  13. Wave propagation and absorption of sandwich beams containing interior dissipative multi-resonators.

    Science.gov (United States)

    Chen, H; Li, X P; Chen, Y Y; Huang, G L

    2017-04-01

    In this study, a sandwich beam with periodic multiple dissipative resonators in the sandwich core material is investigated for broadband wave mitigation and/or absorption. An analytical approach based on the transfer matrix method and Bloch theorem is developed for both infinite and finite sandwich structures. Wave attenuation constants are theoretically obtained to examine the effects of various system parameters on the position, width and wave attenuation performance of the band gaps. The wave absorption coefficient of the sandwich beam is quantitatively studied to distinguish wave attenuation mechanisms caused by reflection and absorption. It is numerically demonstrated that a transient blast-induced elastic wave with broadband frequencies can be almost completely mitigated or absorbed at a subwavelength scale. The results of this study could be used for developing new multifunctional composite materials to suppress impact-induced and/or blast-induced elastic waves which may cause severe local damage to engineering structures.

  14. ELISA法不同检验人员检测HIV抗体结果的比较%Comparison on the results of ELISA method in different inspectors for HIV antibody testing

    Institute of Scientific and Technical Information of China (English)

    吴科明

    2013-01-01

    OBJECTIVE To study the differences between the results of inspection by ELISA which were detected by different inspectors.To reduce the personal error and provide the data to make the lab standard operating procedure (SOP).METHODS Analyzed the same kind of blood serum data with the statistics,which were detected by different inspectors.RESULTS There was no case of false negative,sensitivity and specificity were 100%,precision of 7.64%-14.02%,and the total mean of each group was unequal.CONCLUSION The differences between the inspectors are remarkable,need to strengthen and regulate various aspects in order to reduce human error and individual differences.%目的 了解ELISA法不同检验人员检测HIV抗体结果间的差异,为减少人为误差,进一步规范实验室标准操作程序(SOP)提供数据.方法 对不同检验人员检测出的同一组血清数据运用统计学方法分析.结果 没有一例假阴性,灵敏度和特异性均为100%,精密度为7.64%~14.02%,各组数据总体均数不相等.结论 不同检验人员之间的个体差异是很显著,需要从多方面加强及规范,以减少人为误差及个体间的差异.

  15. A new ELISA for determination of potency in snake antivenoms.

    Science.gov (United States)

    Rial, A; Morais, V; Rossi, S; Massaldi, H

    2006-09-15

    A competitive ELISA for potency determination of bothropic equine antivenom was developed and compared to the conventional in vivo ED(50) assay, with the aim of partially substituting the in vivo assay in the monitoring of antivenom immunoglobulin levels. On this purpose, blood samples were taken at different times during and after the immunization protocol of the lot of horses used for production of snake antivenom at the Instituto de Higiene, Uruguay. Both the competitive ELISA and the ED(50) assay were performed on those samples. In addition, a group of five commercial pepsin-digested antivenoms were tested by both methods. A significant (P<0.001) correlation (Pearson's r=0.957) was found between the ELISA titres and the corresponding ED(50) values, indicating that the in vitro test can estimate the neutralizing antibody capacity of the sera as well as the in vivo assay. By means of this new ELISA, it was found that the immunized animals maintained good venom antibody titres, in the order of 20-50% of the maximum achieved, even 10 month after the end of the immunization schedule. The main advantage of our ELISA design is its ability to correctly estimate the neutralization capacity of crude hyperimmune plasma and antivenom sera independently of their antibody composition in terms of whole IgG or F(ab')(2) fragment.

  16. On the Milnor fibers of sandwiched singularities

    OpenAIRE

    Nemethi, Andras; Popescu-Pampu, Patrick

    2009-01-01

    The sandwiched surface singularities are those rational surface singularities which dominate birationally smooth surface singularities. de Jong and van Straten showed that one can reduce the study of the deformations of a sandwiched surface singularity to the study of deformations of a 1-dimensional object, a so-called decorated plane curve singularity. In particular, the Milnor fibers corresponding to their various smoothing components may be reconstructed up to diffeomorphisms from those de...

  17. A Sandwich Electrochemical Immunosensor Using Magnetic DNA Nanoprobes for Carcinoembryonic Antigen

    Directory of Open Access Journals (Sweden)

    Ning Gan

    2011-10-01

    Full Text Available A novel magnetic nanoparticle-based electrochemical immunoassay of carcinoembryonic antigen (CEA was designed as a model using CEA antibody-functionalized magnetic beads [DNA/Fe3O4/ZrO2; Fe3O4 (core/ZrO2 (shell nano particles (ZMPs] as immunosensing probes. To design the immunoassay, the CEA antibody and O-phenylenediamine (OPD were initially immobilized on a chitosan/nano gold composite membrane on a glassy carbon electrode (GCE/CS-nano Au, which was used for CEA recognition. Then, horseradish peroxidase (HRP-labeled anti-CEA antibodies (HRP-CEA Ab2 were bound to the surface of the synthesized magnetic ZMP nanoparticles as signal tag. Thus, the sandwich-type immune complex could be formed between secondary antibody (Ab2 modified DNA/ZMPs nanochains tagged by HRP and GCE/CS-nano Au. Unlike conventional nanoparticle-based electrochemical immunoassays, the recognition elements of this immunoassay included both electron mediators and enzyme labels, which obviously simplifies the electrochemical measurement process. The sandwich-type immunoassay format was used for online formation of the immunocomplex of CEA captured in the detection cell with an external magnet. The electrochemical signals derived from HRP during the reduction of H2O2 with OPD as electron mediator were measured. The method displayed a high sensitivity for CEA detection in the range of 0.008–200 ng/mL, with a detection limit of 5 pg/mL (estimated at a signal-to-noise ratio of 3. The precision, reproducibility, and stability of the immunoassay were good. The use of the assay was evaluated with clinical serum samples, and the results were in excellent accordance with those obtained using the standard enzyme-linked immunosorbent assay (ELISA method. Thus, the magnetic nanoparticle-based assay format is a promising approach for clinical applications, and it could be further developed for the detection of other biomarkers in cancer diagnosis.

  18. Impact load mitigation in sandwich beams using local resonators

    CERN Document Server

    Sharma, B

    2015-01-01

    Dynamic response of sandwich beams with resonators embedded in the cores subjected to impact loads is studied. Using finite element models the effectiveness of various local resonator frequencies under a given impact load is compared to the behavior of an equivalent mass beam. It is shown that addition of appropriately chosen local resonators into the sandwich beam is an effective method of improving its flexural bending behavior under impact loads. The effect of a given local resonance frequency under different impact load durations is also studied. It is demonstrated that the choice of appropriate local resonance frequency depends on the impact duration. Further, by performing transverse impact experiments, the finite element models are verified and the advantage of using internal resonators under impact loading conditions is demonstrated.

  19. Non-linear Behavior of Curved Sandwich Panels

    DEFF Research Database (Denmark)

    Berggreen, Carl Christian; Jolma, P.; Karjalainen, J. P.;

    2003-01-01

    In this paper the non-linear behavior of curved sandwich panels is investigated both numerically and experimentally. Focus is on various aspects of finite element modeling and calculation procedures. A simply supported, singly curved, CFRP/PVC sandwich panel is analyzed under uniform pressure load...... and results are compared to test data. A novel test arrangement utilizing a water filled cushion to create the uniform pressure load on curved panel specimen is used to obtain the experimental data. The panel is modeled with three different commercial finite element codes. Two implicit and one explicit code...... are used with various element types, modeling approaches and material models. The results show that the theoretical and experimental methods generally show fair agreement in panel non-linear behavior before collapse. It is also shown that special attention to detail has to be taken, because the predicted...

  20. Applications of thin-film sandwich crystallization platforms

    Energy Technology Data Exchange (ETDEWEB)

    Axford, Danny, E-mail: danny.axford@diamond.ac.uk; Aller, Pierre; Sanchez-Weatherby, Juan; Sandy, James [Diamond Light Source, Harwell Oxford, Didcot OX11 0DE (United Kingdom)

    2016-03-24

    Crystallization via sandwiches of thin polymer films is presented and discussed. Examples are shown of protein crystallization in, and data collection from, solutions sandwiched between thin polymer films using vapour-diffusion and batch methods. The crystallization platform is optimal for both visualization and in situ data collection, with the need for traditional harvesting being eliminated. In wells constructed from the thinnest plastic and with a minimum of aqueous liquid, flash-cooling to 100 K is possible without significant ice formation and without any degradation in crystal quality. The approach is simple; it utilizes low-cost consumables but yields high-quality data with minimal sample intervention and, with the very low levels of background X-ray scatter that are observed, is optimal for microcrystals.

  1. A Simple ELISA Exercise for Undergraduate Biology.

    Science.gov (United States)

    Baker, William P.; Moore, Cathy R.

    Understanding of immunological techniques such as the Enzyme Linked Immuno Sorbent Assay (ELISA) is an important part of instructional units in human health, developmental biology, microbiology, and biotechnology. This paper describes a simple ELISA exercise for undergraduate biology that effectively simulates the technique using a paper model.…

  2. Size Effects in Impact Damage of Composite Sandwich Panels

    Science.gov (United States)

    Dobyns, Alan; Jackson, Wade

    2003-01-01

    Panel size has a large effect on the impact response and resultant damage level of honeycomb sandwich panels. It has been observed during impact testing that panels of the same design but different panel sizes will show large differences in damage when impacted with the same impact energy. To study this effect, a test program was conducted with instrumented impact testing of three different sizes of sandwich panels to obtain data on panel response and residual damage. In concert with the test program. a closed form analysis method was developed that incorporates the effects of damage on the impact response. This analysis method will predict both the impact response and the residual damage of a simply-supported sandwich panel impacted at any position on the panel. The damage is incorporated by the use of an experimental load-indentation curve obtained for the face-sheet/honeycomb and indentor combination under study. This curve inherently includes the damage response and can be obtained quasi-statically from a rigidly-backed specimen or a specimen with any support conditions. Good correlation has been obtained between the test data and the analysis results for the maximum force and residual indentation. The predictions can be improved by using a dynamic indentation curve. Analyses have also been done using the MSC/DYTRAN finite element code.

  3. Application of immunomagnetic particles to enzyme-linked immunosorbent assay (ELISA) for improvement of detection sensitivity of HCG.

    Science.gov (United States)

    Kuo, Hsiao-Ting; Yeh, Jay Z; Wu, Po-Hua; Jiang, Chii-Ming; Wu, Ming-Chang

    2012-01-01

    This investigation was aimed at using superparamagnetic particles to enzyme-linked immunosorbent assay (SPIO-ELISA) of human chorionic gonadotropin (hCG) to enhance detection sensitivity of hCG. We found that N-(3-dimethyl aminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC) was the best cross-linking reagent to link anti hCG α antibody to superparamagnetic particle (SPIO-anti hCG α antibody immunomagnetic particle). To improve the specificity of the assay, a horse radish peroxidase (HRP)-labeled anti-hCG beta monoclonal antibody was used to detect captured hCG using double antibody sandwich ELISA assay. SPIO-ELISA application to determine hCG increased the sensitivity to 1 mIU/mL, which is a level of sensitivity enabling the diagnosis of pregnancy during the early gestational period.

  4. Serial follow-up of repeat voluntary blood donors reactive for anti-HCV ELISA

    Directory of Open Access Journals (Sweden)

    Choudhury N

    2011-01-01

    Full Text Available Background : Voluntary non-remunerated repeat blood donors are perceived to be safer than the first time blood donors. This study was planned for follow-up of previous hepatitis C virus (HCV test results of anti-HCV enzyme-linked immunosorbent assay (ELISA reactive repeat blood donors. The aim was to suggest a protocol for re-entry of the blood donors who are confirmed HCV negative by nucleic acid test (NAT and recombinant immunoblot assay (RIBA. A group of repeat voluntary donors were followed retrospectively who became reactive on a cross sectional study and showed HCV reactivity while donating blood regularly. Material and Methods: A total of 51,023 voluntary non remunerated blood donors were screened for anti-HCV ELISA routinely. If anybody showed positivity, they were tested by two ELISA kits (screening and confirmatory and then confirmed infection status by NAT and or RIBA. The previous HCV test results of repeat donors reactive by anti-HCV ELISA were looked back from the records. Data of donors who were repeat reactive with single ELISA kit (in the present study were analyzed separately from those reactive with two ELISA kits (in the present study. Results: In this study, 140 (0.27% donors who were reactive by anti HCV ELISA were included. Out of them, 35 were repeat voluntary donors and 16 (11.43% were reactive with single ELISA kit. All 16 donors were reactive by single ELISA kit occasionally in previous donations. Their present ELISA positive donations were negative for HCV NAT and RIBA. A total of 19 (13.57% donors were reactive with two ELISA kits. In their previous donations, the donors who were reactive even once with two ELISA kits were consistently reactive by the same two ELISA kits in their next donations also. Conclusion: Donor sample reactive by only single ELISA kit may not be considered as infectious for disposal as they were negative by NAT and or RIBA. One time ELISA positivity was found probably due to ELISA kit

  5. A Monoclonal Antibody Based Capture ELISA for Botulinum Neurotoxin Serotype B: Toxin Detection in Food

    Directory of Open Access Journals (Sweden)

    Larry H. Stanker

    2013-11-01

    Full Text Available Botulism is a serious foodborne neuroparalytic disease, caused by botulinum neurotoxin (BoNT, produced by the anaerobic bacterium Clostridium botulinum. Seven toxin serotypes (A–H have been described. The majority of human cases of botulism are caused by serotypes A and B followed by E and F. We report here a group of serotype B specific monoclonal antibodies (mAbs capable of binding toxin under physiological conditions. Thus, they serve as capture antibodies for a sandwich (capture ELISA. The antibodies were generated using recombinant peptide fragments corresponding to the receptor-binding domain of the toxin heavy chain as immunogen. Their binding properties suggest that they bind a complex epitope with dissociation constants (KD’s for individual antibodies ranging from 10 to 48 × 10−11 M. Assay performance for all possible combinations of capture-detector antibody pairs was evaluated and the antibody pair resulting in the lowest level of detection (L.O.D., ~20 pg/mL was determined. Toxin was detected in spiked dairy samples with good recoveries at concentrations as low as 0.5 pg/mL and in ground beef samples at levels as low as 2 ng/g. Thus, the sandwich ELISA described here uses mAb for both the capture and detector antibodies (binding different epitopes on the toxin molecule and readily detects toxin in those food samples tested.

  6. An Analysis of Nondestructive Evaluation Techniques for Polymer Matrix Composite Sandwich Materials

    Science.gov (United States)

    Cosgriff, Laura M.; Roberts, Gary D.; Binienda, Wieslaw K.; Zheng, Diahua; Averbeck, Timothy; Roth, Donald J.; Jeanneau, Philippe

    2006-01-01

    Structural sandwich materials composed of triaxially braided polymer matrix composite material face sheets sandwiching a foam core are being utilized for applications including aerospace components and recreational equipment. Since full scale components are being made from these sandwich materials, it is necessary to develop proper inspection practices for their manufacture and in-field use. Specifically, nondestructive evaluation (NDE) techniques need to be investigated for analysis of components made from these materials. Hockey blades made from sandwich materials and a flat sandwich sample were examined with multiple NDE techniques including thermographic, radiographic, and shearographic methods to investigate damage induced in the blades and flat panel components. Hockey blades used during actual play and a flat polymer matrix composite sandwich sample with damage inserted into the foam core were investigated with each technique. NDE images from the samples were presented and discussed. Structural elements within each blade were observed with radiographic imaging. Damaged regions and some structural elements of the hockey blades were identified with thermographic imaging. Structural elements, damaged regions, and other material variations were detected in the hockey blades with shearography. Each technique s advantages and disadvantages were considered in making recommendations for inspection of components made from these types of materials.

  7. Failure Predictions of Out-of-Autoclave Sandwich Joints with Delaminations Under Flexure Loads

    Science.gov (United States)

    Nordendale, Nikolas A.; Goyal, Vinay K.; Lundgren, Eric C.; Patel, Dhruv N.; Farrokh, Babak; Jones, Justin; Fischetti, Grace; Segal, Kenneth N.

    2015-01-01

    An analysis and a test program was conducted to investigate the damage tolerance of composite sandwich joints. The joints contained a single circular delamination between the face-sheet and the doubler. The coupons were fabricated through out-of-autoclave (OOA) processes, a technology NASA is investigating for joining large composite sections. The four-point bend flexure test was used to induce compression loading into the side of the joint where the delamination was placed. The compression side was chosen since it tends to be one of the most critical loads in launch vehicles. Autoclave cure was used to manufacture the composite sandwich sections, while the doubler was co-bonded onto the sandwich face-sheet using an OOA process after sandwich panels were cured. A building block approach was adopted to characterize the mechanical properties of the joint material, including the fracture toughness between the doubler and face-sheet. Twelve four-point-bend samples were tested, six in the sandwich core ribbon orientation and six in sandwich core cross-ribbon direction. Analysis predicted failure initiation and propagation at the pre-delaminated location, consistent with experimental observations. A building block approach using fracture analyses methods predicted failure loads in close agreement with tests. This investigation demonstrated a small strength reduction due to a flaw of significant size compared to the width of the sample. Therefore, concerns of bonding an OOA material to an in-autoclave material was mitigated for the geometries, materials, and load configurations considered.

  8. Sandwich-like Reconstruction of Anterior Skull Base Defects

    Institute of Scientific and Technical Information of China (English)

    Wang Zheng-min; Wang De-hui

    2001-01-01

    Objective: To evaluate the safety and efficacy of new modality of anterior skull base repair,namely sandwich-like reconstruction of anterior skull base defects. Methods: A retrospective analysis of patients who underwent transcranial or transcranial-facial resections of malignant or benign aggressive tumors involving the anterior skull base was conducted in our department. We used the sandwich-like reconstruction, using pedicled pericranial flap, frontal muscle flap and free abdominal adipose tissue between them, to separate of cranial cavity and aerodigest tract and keep the frontal lobes in place following resections of anterior skull base tumors. Results: From October, 1984 to October, 1998, 116 patients underwent transcranial or transcranialfacial approach for the resection of malignant or aggressive benign tumor, and sandwich-like repairs were performed for the anterior skull base defect. 54 (46.6 % ) patients had previous operation, with a maximum of 5 surgeries. The average age of patients was 35.9 years old, ranging form 6 to 73 years old. Forty-eight (41.4%)patients had malignant neoplasmas, and sixty-eight (58.6%) patients had benign aggressive tumors. In our series, with the maximal follow-ups for as long as 14 years, NO one had early failure of the one-stage reconstruction. CSF fluid leakage was not encountered, nor was ascending bacterial meningitis observed. No immediate or delayed prolapse of dura or frontal lobes was observed. Conclusion: We conclude that the sandwich-like reconstruction, using pericranial flap, frontal muscle flap and free abdominal adipose between them, is an extremely safe and effective procedure for the repair of skull base defect, even when tumor extensively involves anterior skull base.

  9. Sandwich-like Reconstruction of Anterior Skull Base Defects

    Institute of Scientific and Technical Information of China (English)

    WangZheng-min,MD; WangDe-hui,MD

    2001-01-01

    Objective:To evaluate the safety and efficacy of new modality of anterior skull base repair,namely sandwich-like reconstruction of anterior skull base defects. Methods : A retrospective analysis of patients who underwent wanscranial or wanscranial-facial resections of malignant or benign aggressive tumors involving the anterior skull base was conducted in our department. We used the sandwich-like reconstruction, using pedicled pericranial flap, frontal muscle flap and free abdominal adipose tissue between them, to separate of cranial cavity and aerodigest tract and keep the frontal lobes in place following resections of anterior skull base tumors. Results: From October, 1984 to October, 1998, 116 patients underwent tmnscranial or tmnscranial-facial approach for the resection of malignant or aggressive benign tumor, and sandwich-like repairs were performed for the anterior skull base defect.54 (46.6%) patients had previous operation, with a maximum of 5 surgeries. The average age of patients was 35.9 years old, ranging form 6 to 73 years old. Forty-eight (41.4%) patients had malignant neoplasmas, and sixty-eight (58.6%) patients had benign aggressive tumors. In our series, with the maximal follow-ups for as long as 14 years, NO one had early failure of the one-stage reconstruction. CSF fluid leakage was not encountered, nor was ascending bacterial meningitis observed. No immediate or delayed prolapse of dura or frontal lobes was observed. Conclusion: We conclude that the sandwich-like reconstruction, using pericranial flap, frontal muscle flap and free abdominal adipose between them, is an extremely safe and effective procedure for the repair of skull base defect, even when tumor extensively involves anterior skull base.

  10. Impact properties of aluminium - glass fiber reinforced plastics sandwich panels

    Directory of Open Access Journals (Sweden)

    Mathivanan Periasamy

    2012-06-01

    Full Text Available Aluminium - glass fiber reinforced plastics (GFRP sandwich panels are hybrid laminates consisting of GFRP bonded with thin aluminum sheets on either side. Such sandwich materials are increasingly used in airplane and automobile structures. Laminates with varying aluminium thickness fractions, fiber volume fractions and orientation in the layers of GFRP were fabricated by hand lay up method and evaluated for their impact performance by conducting drop weight tests under low velocity impacts. The impact energy required for initiating a crack in the outer aluminium layer as well as the energy required for perforation was recorded. The impact load-time history was also recorded to understand the failure behavior. The damage depth and the damage area were measured to evaluate the impact resistance. Optical photography and scanning electron micrographs were taken to visualize the crack and the damage zone. The bidirectional cross-ply hybrid laminate (CPHL has been found to exhibit better impact performance and damage resistance than the unidirectional hybrid laminate (UDHL. Increase in aluminium thickness fraction (Al tf and fiber volume fraction (Vf resulted in an increase in the impact energy required for cracking and perforation. On an overall basis, the sandwich panels exhibited better impact performance than the monolithic aluminium.

  11. Systematic selection of modified aptamer pairs for diagnostic sandwich assays.

    Science.gov (United States)

    Ochsner, Urs A; Green, Louis S; Gold, Larry; Janjic, Nebojsa

    2014-01-01

    Protein diagnostic applications typically require pairs of analyte-specific reagents for capture and detection. We developed methods for the systematic isolation of slow off-rate modified aptamer (SOMAmer) reagents that bind to different epitopes and allow efficient pair-wise screening of multiple ligands. SOMAmers were generated via a second systematic evolution of ligands by exponential enrichment (SELEX), using complexes of target proteins with a primary, non-amplifiable SOMAmer and employing different modified nucleotides (e.g., naphthylmethyl- or tryptaminocarbonyl-dU) to favor alternate binding epitopes. Non-competing binding of primary and secondary SOMAmers was tested in radiolabel competition and sandwich binding assays. Multiplexed high-throughput screening for sandwich pairs utilized the Luminex platform, with primary SOMAmers as capture agents attached to different types of LumAvidin beads, which were then pooled for testing the secondary SOMAmers individually as detection agents. Functional SOMAmer pairs were obtained for Clostridium difficile binary toxin (CdtA) and for a panel of human proteins (ANGPT2, TSP2, CRDL1, MATN2, GPVI, C7, PLG) that had been previously identified as promising markers for cardiovascular risk. The equilibrium dissociation constants (Kd values) ranged from 0.02-2.7 nM, and the detection limits were in the low picomolar range for these proteins in SOMAmer sandwich assays. These results indicate that SOMAmer pairs hold promise for the development of rapid tests or specific diagnostic panels.

  12. Energy Dissipation in Sandwich Structures During Axial Compression

    DEFF Research Database (Denmark)

    Urban, Jesper

    2002-01-01

    The purpose of this paper is to investigate the energy dissipation in sandwich structures during axial crushing. Axial crushing tests on six sandwich elements are described. The sandwich elements consist of a polyurethane core and E-glass/Polyester skin. The elements compare to full-scale structu......The purpose of this paper is to investigate the energy dissipation in sandwich structures during axial crushing. Axial crushing tests on six sandwich elements are described. The sandwich elements consist of a polyurethane core and E-glass/Polyester skin. The elements compare to full...

  13. Fate of Estrogens in Soils and Detection by ELISA

    Science.gov (United States)

    Caron, Emmanuelle; Sheedy, Claudia; Farenhorst, Annemieke; Zvomuya, Francis; Gaultier, Jeanette; Goddard, Tom

    2010-05-01

    Land application of manure can contribute to the release of estrogenic compounds in the environment. Estrogens may move from soils to water by processes such as runoff and leaching. The objectives of the present study were to determine the fate of estrogens in soils and to develop a detection method for these compounds. The sorption (soil sorption coefficient (Kd) and sorption coefficient per unit organic carbon (Koc)) of 17β-estradiol, estrone, estriol and equol were studied, using batch equilibrium experiments, in 121 surface soils from Alberta, Canada. The mineralization of [4-14C] 17β-estradiol was determined in soil microcosms in a subset of 36 samples. Quantitative relationships at the regional level were explored using partial least squares regression (PLS) (between Kd or Koc values and soil properties) and by ordinary least squares regression (between Kd or Koc values of different estrogens). Soil properties (r2 0.51-0.87 for Kd and 0.32-0.44 for Koc) provided better prediction models than using the data of different estrogens (r2 0.38-0.71 for Kd and 0.18-0.40 for Koc). PLS regression models for mineralization parameters of 17ß-estradiol had lower predictive power (lower r2)than models developed for sorption parameters. In addition, it has become of primary importance to develop sensitive detection methods that are able to detect low estrogen concentrations (ng L-1) in a wide variety of environmental matrices in order to validate the prediction of their fate and to study their presence in affected ecosystems. Conjugates were synthesized using a mixed anhydride reaction and two Enzyme-Linked Immunosorbent Assays (ELISAs) were developed using polyclonal antibodies. One ELISA was highly specific for 17β-estradiol (with an IC50 of 243 ng mL-1) and the second allowed for the broader detection of 17β-estradiol, estrone and estriol (with an IC50 of 18 ng mL-1 for 17β-estradiol). The cross-reactivity of both ELISAs was studied against 13 compounds (natural

  14. An h-p Finite Element Vibration Analysis of Open Conical Sandwich Panels and Conical Sandwich Frusta

    Science.gov (United States)

    BARDELL, N. S.; LANGLEY, R. S.; DUNSDON, J. M.; AGLIETTI, G. S.

    1999-09-01

    The vibration study of a general three-layer conical sandwich panel based on theh -p version of the finite element method is presented in this paper. No restriction is placed on the degree of curvature of the shell, thereby relaxing the strictures associated with shallow shell theory. The methodology incorporates a new set of trigonometric functions to provide the element p -enrichment, and elements may be joined together to model either open conical panels, or complete conical frusta (circumferentially connected, but open at each end). The full range of classical boundary conditions, which includes free, clamped, simply supported and shear diaphragm edges, may be applied in any combination to open and closed panels, thereby facilitating the study of a wide range of conical sandwich shells. The convergence properties of this element have been established for different combinations of the h - and p -parameters, thereby assuring its integrity for more general use. Since very little work has been reported on the vibration characteristic of either circumferentially closed or open conical sandwich panels, the main thrust of this work has been to present and validate an efficient modelling technique, rather than to perform numerous parameter and/or sensitivity studies. To this end, some new results are presented and subsequently validated using a commercially available finite element package. It is shown that for results of comparable accuracy, models constructed using the h-p formulation require significantly fewer degrees of freedom than those assembled using the commercial package. Some preliminary experimental results are also included for completeness.

  15. Sandwich Panels Evaluated With Ultrasonic Spectroscopy

    Science.gov (United States)

    Cosgriff, Laura M.

    2004-01-01

    Enhanced, lightweight material systems, such as 17-4PH stainless steel sandwich panels are being developed for use as fan blades and fan containment systems for next-generation engines. The bond strength between the core and face sheets is critical in maintaining the structural integrity of the sandwich structure. To improve the inspection and production of these systems, researchers at the NASA Glenn Research Center are using nondestructive evaluation (NDE) techniques, such as ultrasonic spectroscopy, to evaluate the brazing quality between the face plates and the metallic foam core. The capabilities and limitations of a swept-frequency approach to ultrasonic spectroscopy were evaluated with respect to these sandwich structures. This report discusses results from three regions of a sandwich panel representing different levels of brazing quality between the outer face plates and a metallic foam core. Each region was investigated with ultrasonic spectroscopy. Then, on the basis of the NDE results, three shear specimens sectioned from the sandwich panel to contain each of these regions were mechanically tested.

  16. Development of an ELISA Method for Multi-Residue Detecting of Fluoroquinolones%氟喹诺酮类药物多残留酶联免疫检测方法的建立

    Institute of Scientific and Technical Information of China (English)

    李新朋; 姜金庆; 钱爱东; 王自良; 范国英; 单晓峰; 康元环; 李艺

    2014-01-01

    释;反应温度均为37℃,标准品与单抗同时加入反应15 min,洗板后加二抗反应25 min;显色10 min,2H5细胞株腹水敏感性和广谱特异性最好,2H5的线性回归方程为y=-28.022x+56.219,R2=0.9782,对10种FQs的IC50值分别为环丙沙星(CPFX)1.67 ng·mL-1、诺氟沙星(NOR)1.82 ng·mL-1、培氟沙星(PEF)1.97 ng·mL-1、恩诺沙星(ENR)1.54 ng·mL-1、单诺沙星(DAN)2.79 ng·mL-1、洛美沙星(LOM)3.38 ng·mL-1、氧氟沙星(OFL)5.50 ng·mL-1、麻保沙星(MAR)4.40 ng·mL-1、沙拉沙星(SAR)11.76 ng·mL-1、二氟沙星(DIF)13.60 ng·mL-1,与10种FQs的交叉反应率为12.3%—108.4%,与非 FQs交叉反应率均低于0.01%,对10种FQs的检测限(LODs)为0.09 ng·mL-1—0.64 ng·mL-1。icELISA法鸡肉中10种FQs的回收率为80.5%—91.8%,HPLC法的回收率为85.1%—95.7%,二者变异系数均低于10.0%;两种检测方法差异不显著(P>0.05)。【结论】该试验获得了高效价、敏感、广谱特异性的mAbs,建立的icELISA方法可用于同时检测鸡肉中10种FQs。%Objective]Fluoroquinolones(FQs) are widely used in veterinary medicine for the treatment and prevention of bacterial infection. With the increasing use, FQs residues in animal edible tissues have caused serious public health problems and attracted serious attention by research scholars all over the world. The objective of this study was to produce class-specific monoclonal antibodies (mAbs) against fluoroquinolones (FQs), establish competitive indirect enzyme linked immunesorbent assay (icELISA), and in order to lay a foundation for detection of multi-residue FQs in animal foods.[Method]The aminobutyric acid was introduced to carboxyl of ciprofloxacin as hapten (CPFX-A) and was proved by (+) ESI-MS spectrum, which was conjugated to bovine serum albumin (BSA) as immunogen (CPFX-A-BSA) by the N,N'-Dicyclohexylcarbodiimide (DDC) method, and to ovalbumin (OVA) as coating antigen(CPFX-A-OVA) by

  17. Herpes simplex virus type 2 antibody detection performance in Kisumu, Kenya, using the Herpeselect ELISA, Kalon ELISA, Western blot and inhibition testing.

    Science.gov (United States)

    Smith, J S; Bailey, R C; Westreich, D J; Maclean, I; Agot, K; Ndinya-Achola, J O; Hogrefe, W; Morrow, R A; Moses, S

    2009-04-01

    In certain parts of Africa, type-specific herpes simplex virus type 2 (HSV-2) ELISAs may have limited specificity. To date, no study has been conducted to validate HerpeSelect and Kalon type-specific HSV-2 ELISAs using both the Western blot and recombinant gG ELISA inhibition testing as reference standards. A total of 120 men who were HIV seronegative (aged 18-24 years) provided blood samples. HSV-2 IgG serum antibodies were detected using four different methods: HerpeSelect HSV-2 ELISA (n = 120), Kalon HSV-2 ELISA (n = 120), University of Washington Western blot (n = 101) and a recombinant inhibition test (n = 93). HSV-2 seroprevalence differed significantly by HSV-2 detection method, ranging from 24.8% with the Western blot to 69.8% with the HerpeSelect ELISA. Using the Western blot as the reference standard, the HerpesSelect had the highest sensitivity for HSV-2 antibody detection (100%) yet lowest specificity (40%). Similar results were obtained using the inhibition test as the reference standard. The sensitivity and specificity of the Kalon test versus the Western blot were 92% and 79%, respectively, and 80% and 82% versus the inhibition test. Using the inhibition test as the reference standard, the sensitivity of the Western blot appeared low (49%). In men in western Kenya who were HIV seronegative, the HerpeSelect and Kalon type-specific ELISAs had high sensitivities yet limited specificities using the Western blot as reference standard. Overall, the Kalon ELISA performed better than the HerpeSelect ELISA in these young men from Kisumu. Further understanding is needed for the interpretation of HSV-2 inhibition or ELISA test positive/ Western blot seronegative results. Before HSV-2 seropositivity may be reliably reported in selected areas of Africa, performance studies of HSV-2 serological assays in individual geographical areas are recommended.

  18. ELISA for the serology of FIP virus.

    NARCIS (Netherlands)

    A.D.M.E. Osterhaus (Ab); A. Kroon; R.M.S. Wirahadiredja

    1979-01-01

    textabstractAn enzyme linked immunosorbent assay (ELISA) for feline infectious peritonitis (FIP) virus serology is described. The assay is analogous to a previously developed indirect heterologous immunofluorescence test (IFT) in which transmissible gastroenteritis (TGE) viral antigen was used. Comp

  19. A sandwich enzyme-linked immunosorbent assay for the quantification of insoluble membrane and scaffold proteins.

    Science.gov (United States)

    Geumann, Constanze; Grønborg, Mads; Hellwig, Michaela; Martens, Henrik; Jahn, Reinhard

    2010-07-15

    Enzyme-linked immunosorbent assays (ELISAs) are applied for the quantification of a vast diversity of small molecules. However, ELISAs require that the antigen is present in a soluble form in the sample. Accordingly, the few ELISAs described so far targeting insoluble proteins such as integral membrane and scaffold proteins have been restricted by limited extraction efficiencies and the need to establish an individual solubilization protocol for each protein. Here we describe a sandwich ELISA that allows the quantification of a diverse array of synaptic membrane and scaffold proteins such as munc13-1, gephyrin, NMDA R1 (N-methyl-d-aspartate receptor subunit 1), synaptic vesicle membrane proteins, and SNAREs (soluble N-ethylmaleimide-sensitive factor attachment protein receptors). The assay is based on initial solubilization by the denaturing detergent sodium dodecyl sulfate (SDS), followed by partial SDS removal using the detergent Triton X-100, which restores antigenicity while keeping the proteins in solution. Using recombinant standard proteins, we determined assay sensitivities of 78ng/ml to 77pg/ml (or 74-0.1fmol). Calibration of the assay using both immunoblotting and mass spectroscopy revealed that in some cases correction factors need to be included for absolute quantification. The assay is versatile, allows parallel processing and automation, and should be applicable to a wide range of hitherto inaccessible proteins.

  20. Vibro-acoustics of lightweight sandwich structures

    CERN Document Server

    Lu, Tianjian

    2014-01-01

    Vibro-Acoustics of Lightweight Sandwich Structures introduces the study of the coupled vibration and acoustic behavior of lightweight sandwich structures in response to harmonic force and sound pressure. This book focuses on the theoretical modeling and experimental investigation of lightweight sandwich structures in order to provide a predictive framework for vibro-acoustic characteristics of typical engineering structures. Furthermore, by developing solution tools, it concentrates on the influence of key systematic parameters leading to effective guidance for optimal structure design toward lightweight, high-stiffness and superior sound insulation capability. This book is intended for researchers, scientists, engineers and graduate students in mechanical engineering especially in structural mechanics, mechanics and acoustics. Fengxian Xin and Tianjian Lu both work at the School of Aerospace, Xi’an Jiaotong University.

  1. Immobilized carboxymethylated dextran coatings for enhanced ELISA.

    Science.gov (United States)

    Liberelle, Benoît; Merzouki, Abderrazzak; De Crescenzo, Gregory

    2013-03-29

    We here report the development of a new generation of enzyme-linked immunosorbent assay (ELISA) that takes advantage of a low-fouling carboxymethylated dextran (CMD) layer chemically grafted on ELISA wells. In our approach, the overnight capture antibody adsorption step found in classical ELISA was replaced by a covalent attachment step to the CMD layer completed in 15 min. As a model, the potential of our approach was highlighted using commercially available anti-human epidermal growth factor (EGF) antibodies to quantify EGF present in various samples. Of interest, the grafted CMD layer was found to be as efficient as the commonly used bovine serum albumine (BSA) to reduce non-specific adsorption, thus eliminating the need of a time-consuming BSA blocking step normally required in classical ELISA. Our results demonstrated similar specificity, affinity, and intra- and inter-assay variations regardless of the diluent used in the assay (BSA-based diluent or protein-free buffer solution) when compared to standard ELISA. Finally, accuracy and precision of the CMD-based ELISA were verified by a spike and recovery test. Dilutions of recombinant human EGF in serum from healthy human volunteers showed almost-perfect linearity and mean recovery rates ranging between 90 and 110%.

  2. 狂犬病病毒抗体竞争ELISA检测方法的建立及初步应用%Development and Preliminary Application of Competitive ELISA Method for Rabies Virus Antibody

    Institute of Scientific and Technical Information of China (English)

    王伟伟; 王远征; 张国强; 马昱

    2011-01-01

    Objective To develop a competitive ELBA method for rabies virus (RV) antibody and use for determination of antibody levels after immunization with of rabies vaccines prepared from various virus strains. Methods NIH mice were immunized with the antigens of commercial rabies vaccine prepared with various strains, and the obtained immune sera were determined for antibody titer by using the EIA plate coated with the antigens of various vaccine strains. The antigen diversity was analyzed by cross neutralization test of antigens and antibodies of various strains, based on which the EIA plate was coated with antigens of various strains mixed at different ratios to determine positive serum samples, and the results were compared with those by rapid fluorescent focus inhibition test (RFFTT) to determine the coating antigen, and a competitive ELISA method for RV antibody was developed, of which the optimal quantitative detection range and sensitivity were determined by plotting a standard curve, the cut-off value was determined, and the specificity, precision, accuracy and stability were verified. Serum samples were determined by the developed method, and the results were compared with those by other commercial kits for RV antibody to analyze the consistence and correlation. Results The coating pattern was optimized as mixing the antigens from AG and CTN-1 strains at a ratio of 4 : 1. After optimization, the linear detection range of the developed kit for antibody titer was 535. 00 - 33. 44 mlU/ml (r > 0. 99), while the minimum detection limit and cut-off value were 8. 36 mlU/ml and 0. 735 mill respectively. All the detection results of human serum albumin, tetanus positive serum, HBsAg positive serum and diphtheria positive quality control serum by the developed method were negative. The inter-variation coefficient of detection result by the developed method was 6. 68% ~ 7. 84%, while the recovery rate was 97. 25% ~ 104. 50%. The detection results by the developed kit after

  3. A Conformal Thin-Sandwich Solver for Generic Initial Data

    CERN Document Server

    East, William E; Pretorius, Frans

    2012-01-01

    We present a new scheme for constructing initial data for the Einstein field equations using the conformal thin-sandwich formulation that does not assume conformal flatness or approximate Killing vectors. This includes a method for determining free data based on superposition, as well as a way to handle black hole singularities without excision. We numerically solve the constraint equations using a multigrid algorithm with mesh refinement. We demonstrate the efficacy of the method with initial data solutions for several applications: a quasi-circular binary black hole merger, a dynamical capture black hole-neutron star merger, and an ultrarelativistic collision.

  4. A rapid sandwich immunoassay for human fetuin A using agarose-3-aminopropyltriethoxysilane modified microtiter plate.

    Science.gov (United States)

    Vashist, Sandeep Kumar; Schneider, E Marion; Luong, John H T

    2015-07-09

    A rapid sandwich immunoassay (IA) with enhanced signal response for human fetuin A (HFA) was developed by modifying the surface of a KOH-treated polystyrene microtiter plate (MTP) with agarose and 3-aminopropyltriethoxysilane (APTES). The agarose-APTES complex binds covalently to the hydroxyl moiety of the MTP plate to serve as a binding platform for bioconjugation of EDC-activated anti-HFA antibody (Ab) via carbodiimide coupling. The one-step kinetics-based sandwich enzyme-linked immunosorbent assay (ELISA) enabled the detection of HFA in 30 min with a limit of detection (LOD) and a linear range of 0.02 ng mL(-1) and 1-243 ng mL(-1), respectively. It detected HFA spiked in diluted human whole blood and serum, and HFA in ethylenediaminetetraacetic acid (EDTA)-plasma of patients with high precision similar to that of conventional ELISA. The anti-HFA Ab-bound agarose-functionalized MTPs retained their functional activity after 6 weeks of storage in 0.1 M PBS, pH 7.4 at 4 °C.

  5. Quantification of human tissue transglutaminase by a luminescence sandwich enzyme-linked immunosorbent assay.

    Science.gov (United States)

    Wolf, Johannes; Lachmann, Ingolf; Wagner, Uta; Osman, Awad A; Mothes, Thomas

    2011-12-15

    Tissue transglutaminase (tTG) is a calcium-dependent enzyme that catalyzes crosslinking of peptidic glutamine residues with primary amines via isopeptide bonds and hydrolysis of ATP or GTP. The enzyme exerts a variety of functions at the cellular and tissue levels that may be disturbed in disease. Its role in pathoprocesses is poorly understood. For investigation of the involvement of tTG in disease, sensitive and specific assays should be available. We have developed the first sandwich enzyme-linked immunosorbent assay (ELISA) based on two monoclonal antibodies (mabs) against human tTG. tTG is captured by mab 3C10 and detected by biotinylated mab 10F3. After incubation with peroxidase-conjugated streptavidin, bound tTG is visualized by peroxidase reaction applying a luminescence substrate. The detection limit was 40 pg/ml. The assay was highly reproducible. Recovery of spiked tTG in crude samples was greater than 92%. The enzyme could be detected in cellular lysates and tissue homogenates of humans. The effect of typical effectors (retinoic acid and interferon-γ) on tTG expression could be demonstrated. A low signal was also obtained in mice samples, suggesting cross-reactivity of the mabs with murine tTG. The new sandwich ELISA may be successfully applied for investigation of physiological functions of tTG and of disorders associated with inadequate tTG expression.

  6. AA, sandwich line with magnetic horn

    CERN Multimedia

    1980-01-01

    The magnetic horn, focusing the antiprotons emanating from the target, was affixed to a sandwich line through which the 150 kA pulses were supplied. Expecting to have to change from time to time the fragile horn (inner conductor only 0.7 mm thick), the assembly was designed for quick exchange. At the lower end of the sandwich line we see the connectors for the high-current cables, at the upper end the magnet horn. It has just been lifted from the V-supports which held it aligned downstream of the target. Continue with 8010293.

  7. Nonlinear dynamic analysis of sandwich panels

    Science.gov (United States)

    Lush, A. M.

    1984-01-01

    Two analytical techniques applicable to large deflection dynamic response calculations for pressure loaded composite sandwich panels are demonstrated. One technique utilizes finite element modeling with a single equivalent layer representing the face sheets and core. The other technique utilizes the modal analysis computer code DEPROP which was recently modified to include transverse shear deformation in a core layer. The example problem consists of a simply supported rectangular sandwich panel. Included are comparisons of linear and nonlinear static response calculations, in addition to dynamic response calculations.

  8. Advanced Mechanical Testing of Sandwich Materials

    DEFF Research Database (Denmark)

    Hayman, Brian; Berggreen, Christian; Jenstrup, Claus

    2008-01-01

    An advanced digital optical system has been used to measure surface strains on sandwich face and core specimens tested in a project concerned with improved criteria for designing sandwich X-joints. The face sheet specimens were of glass reinforced polyester and were tested in tension. The core...... specimens were of PVC foam and were tested in compression. The tests were performed in order to validate the use of the measurement system on these materials and to obtain material data for use in numerical simulations. While some limitations were identified, the optical system performed well and appears...

  9. Overall Buckling and Wringkling of Debonded Sandwich Beams: Finite Element and Experimental Results

    Directory of Open Access Journals (Sweden)

    Bambang K. Hadi

    2006-05-01

    Full Text Available Overall buckling and wrinkling of debonded sandwich beams under compressive loads were analyzed by both finite element and experimental methods. In the finite element method, a quarter and a half models of the specimens were analyzed. It shows that a quarter model is not adequate to analyze buckling of debonded sandwich beams, since it will disregard overall buckling mode that may occur in sandwich beams having compressive loads. At least a half model should be used to analyze buckling of sandwich beams. A finite element program UNA was used extensively to analyze the buckling loads. Experimental buckling of sandwich beams was carried out using a compression testing machine. Two LVDTs were used to measure deflections of the specimen during experimental loading. The loads were measured using load cells available in the machine. Specimens having core thickness of 45 and 75 mm were tested to represent overall and wrinkling modes respectively. The delamination lengths were 20, 60 and 80 mm, which represent 10, 30 and 40% of the beam length. The results show that the differences between experimental and finite element methods were less than 10%. Both overall buckling and wrinkling modes were shown in these specimens.

  10. Development and application of triple antibody sandwich enzyme-linked immunosorbent assays for begomovirus detection using monoclonal antibodies against Tomato yellow leaf curl Thailand virus.

    Science.gov (United States)

    Seepiban, Channarong; Charoenvilaisiri, Saengsoon; Warin, Nuchnard; Bhunchoth, Anjana; Phironrit, Namthip; Phuangrat, Bencharong; Chatchawankanphanich, Orawan; Attathom, Supat; Gajanandana, Oraprapai

    2017-05-30

    Tomato yellow leaf curl Thailand virus, TYLCTHV, is a begomovirus that causes severe losses of tomato crops in Thailand as well as several countries in Southeast and East Asia. The development of monoclonal antibodies (MAbs) and serological methods for detecting TYLCTHV is essential for epidemiological studies and screening for virus-resistant cultivars. The recombinant coat protein (CP) of TYLCTHV was expressed in Escherichia coli and used to generate MAbs against TYLCTHV through hybridoma technology. The MAbs were characterized and optimized to develop triple antibody sandwich enzyme-linked immunosorbent assays (TAS-ELISAs) for begomovirus detection. The efficiency of TAS-ELISAs for begomovirus detection was evaluated with tomato, pepper, eggplant, okra and cucurbit plants collected from several provinces in Thailand. Molecular identification of begomoviruses in these samples was also performed through PCR and DNA sequence analysis of the CP gene. Two MAbs (M1 and D2) were generated and used to develop TAS-ELISAs for begomovirus detection. The results of begomovirus detection in 147 field samples indicated that MAb M1 reacted with 2 begomovirus species, TYLCTHV and Tobacco leaf curl Yunnan virus (TbLCYnV), whereas MAb D2 reacted with 4 begomovirus species, TYLCTHV, TbLCYnV, Tomato leaf curl New Delhi virus (ToLCNDV) and Squash leaf curl China virus (SLCCNV). Phylogenetic analyses of CP amino acid sequences from these begomoviruses revealed that the CP sequences of begomoviruses recognized by the narrow-spectrum MAb M1 were highly conserved, sharing 93% identity with each other but only 72-81% identity with MAb M1-negative begomoviruses. The CP sequences of begomoviruses recognized by the broad-spectrum MAb D2 demonstrated a wider range of amino acid sequence identity, sharing 78-96% identity with each other and 72-91% identity with those that were not detected by MAb D2. TAS-ELISAs using the narrow-specificity MAb M1 proved highly efficient for the detection of

  11. DETECTION OF SERIAL SERUM P-185 LEVEL IN PATIENTS WITH MALIGNANCY USING ELISA

    Institute of Scientific and Technical Information of China (English)

    ZHANG Zhi-hui; WU Ping; LI Li; MIAO Yi-meng; LI Jun

    2006-01-01

    Objective: To investigate P-185 contents in serum of the normal person and cancer patients and it's the significance on prognosis of diseases. Methods: We used ELISA method to evaluate P-185 levels in 193 normal persons, 133 malignancies and 34 hepatocirrhosises were evaluated using ELISA method. Results: Normal person had lower expression of P-185. However, malignancy and hepatocirrhosis patients had a significantly higher expression level of P-185 than normal (P<0.05). Conclusion: ELISA method is an easy and reliable way to measure the level of P-185 in serum. Being a cancer marker, P-185 overexpression can be used for early diagnosis and prognosis of cancer patients.

  12. ELISA for complexes between urokinase-type plasminogen activator and its receptor in lung cancer tissue extracts

    DEFF Research Database (Denmark)

    de Witte, H; Pappot, H; Brünner, N

    1997-01-01

    A sandwich-type ELISA has been developed for the assessment of complexes between urokinase-type plasminogen activator (uPA) and its receptor (uPAR) in extracts of squamous cell lung carcinomas. The assay is based on a combination of rabbit polyclonal anti-uPA antibodies and a biotinylated mouse a......PA:uPAR complexes in lung tumor tissue as well as other types of cancer.......A sandwich-type ELISA has been developed for the assessment of complexes between urokinase-type plasminogen activator (uPA) and its receptor (uPAR) in extracts of squamous cell lung carcinomas. The assay is based on a combination of rabbit polyclonal anti-uPA antibodies and a biotinylated mouse...... indicate that de novo complex formation is a major factor to consider and that complexes analyzed in the presence of this antagonist represent original uPA:uPAR complexes present prior to tumor tissue processing. The present ELISA appears suitable for studying the potential prognostic impact of u...

  13. CUBLIC SPLINE SOLUTIONS OF AXISYMMETRICAL NONLINEAR BENDING AND BRCKLING OF CIRCULAR SANDWICH PLATES

    Institute of Scientific and Technical Information of China (English)

    侯朝胜; 张守恺; 林锋

    2005-01-01

    Cubic B-spline taken as trial function, the nonlinear bending of a circular sandwich plate was calculated by the method of point collocation. The support could be elastic. A sandwich plate was assumed to be Reissner model. The formulae were developed for the calculation of a circular sandwich plate subjected to polynomial distributed loads,uniformly distributed moments, radial pressure or radial prestress along the edge and their combination. Buckling load was calculated for the first time by nonlinear theory. Under action of uniformly distributed loads, results were compared with that obtained by the power series method. Excellences of the program written by the spline collocation method are wide convergent range, high precision and universal.

  14. STUDY ON DETECTING ANTIBODY OF TUBERCULOSIS IN COW MILK WHEY BY THE METHOD OF BA-Dot-ELISA%BA-Dot-ELISA检测牛结核乳清抗体的研究

    Institute of Scientific and Technical Information of China (English)

    杨玉英; 李士泽; 郭士玲; 赵凯

    2000-01-01

    本研究建立了快速检测牛乳中结核病抗体的BA-Dot-ELISA方法.用该法检测28份结核变反阳性牛乳清,阳性率为78.57%,比常规ELISA(64.28%)高.用该法检测布鲁氏菌病牛乳清无交叉反应,但对5份副结核变反阳性牛乳清出现了一定的交叉反应.

  15. New examples of sandwich gravitational waves and their impulsive limit

    CERN Document Server

    Podolsky, J

    1998-01-01

    Non-standard sandwich gravitational waves are constructed from the homogeneous pp vacuum solution and the motions of free test particles in the space-times are calculated explicitly. They demonstrate the caustic property of sandwich waves. By performing limits to impulsive gravitational wave it is demonstrated that the resulting particle motions are identical regardless of the ''initial'' sandwich.

  16. Non-linear analytical solutions for laterally loaded sandwich plates

    DEFF Research Database (Denmark)

    Riber, Hans Jørgen

    1997-01-01

    This work focuses on the response of orthotropic sandwich composite plates with large deflections due to high lateral loads. The results have special application to the design of ship structures. A geometrical nonlinear theory is outlined, on the basis of the classical sandwich plate theory...... of sandwich plates subjected to high lateral loading. (C) 1997 Published by Elsevier Science Ltd. All rights reserved....

  17. Buckling analysis of curved composite sandwich panels subjected to inplane loadings

    Science.gov (United States)

    Cruz, Juan R.

    1993-01-01

    Composite sandwich structures are being considered for primary structure in aircraft such as subsonic and high speed civil transports. The response of sandwich structures must be understood and predictable to use such structures effectively. Buckling is one of the most important response mechanisms of sandwich structures. A simple buckling analysis is derived for sandwich structures. This analysis is limited to flat, rectangular sandwich panels loaded by uniaxial compression (N(sub x)) and having simply supported edges. In most aerospace applications, however, the structure's geometry, boundary conditions, and loading are usually very complex. Thus, a general capability for analyzing the buckling behavior of sandwich structures is needed. The present paper describes and evaluates an improved buckling analysis for cylindrically curved composite sandwich panels. This analysis includes orthotropic facesheets and first-order transverse shearing effects. Both simple support and clamped boundary conditions are also included in the analysis. The panels can be subjected to linearly varying normal loads N(sub x) and N(sub y) in addition to a constant shear load N(sub xy). The analysis is based on the modified Donnell's equations for shallow shells. The governing equations are solved by direct application of Galerkin's method. The accuracy of the present analysis is verified by comparing results with those obtained from finite element analysis for a variety of geometries, loads, and boundary conditions. The limitations of the present analysis are investigated, in particular those related to the shallow shell assumptions in the governing equations. Finally, the computational efficiency of the present analysis is considered.

  18. A novel indirect ELISA for diagnosis of dengue fever

    Directory of Open Access Journals (Sweden)

    Rohan Narayan

    2016-01-01

    Full Text Available Background & objectives: Dengue fever (DF is associated with significant morbidity and mortality in the tropical and sub-tropical regions of the world. Since there are no effective antiviral drugs for treatment, clinicians often rely on the accurate diagnosis of dengue fever to begin supportive therapy at early stages of the illness. The objective of this study was to develop an in-house dengue virus serotype 2 (DENV-2 non-structural protein- 5 (NS5 based indirect ELISA. Methods: DENV-2 was raised in Vero cells and the viral proteins were separated and subsequently the NS5 protein was eluted. Serum samples from primary and secondary dengue fever patients; and acute and convalescent samples from Japanese encephalitis (JE and West Nile virus (WNV cases were used to validate the ELISA. Results: The assay was found to be 100 per cent specific in detecting DENV-2 specific antibodies from patient′s serum. However, in terms of sensitivity, the assay could detect IgM antibodies only from 90 per cent of the primary dengue samples. The IgM/IgG ratio of the primary and secondary samples was 7.24 and 0.64, respectively. Interpretation & conclusions: The results indicate that the DENV-2 NS5 ELISA is dengue group specific and can be used to differentiate dengue infection from other circulating Flavivirus infections. This NS5 ELISA can also be used to distinguish between primary and secondary dengue fever on the basis of IgM/IgG ratios. Further studies with larger sample sizes and different DENV serotypes are required to validate the ELISA.

  19. AA, sandwich line with magnetic horn

    CERN Document Server

    1980-01-01

    Continuation from 8010293: Finally, the sandwich line with the horn is placed on the ground, for the horn to be inspected and, if needed, exchanged for a new one. The whole procedure was trained with several members of the AA team, for quick and safe handling, and to share the radiation dose amongst them.

  20. Proof of the Thin Sandwich Conjecture

    CERN Document Server

    Bartnik, R; Bartnik, Robert; Fodor, Gyula

    1993-01-01

    We prove that the Thin Sandwich Conjecture in general relativity is valid, provided that the data $(g_{ab},\\dot g_{ab})$ satisfy certain geometric conditions. These conditions define an open set in the class of possible data, but are not generically satisfied. The implications for the ``superspace'' picture of the Einstein evolution equations are discussed.

  1. Structural detailing of openings in sandwich panels

    NARCIS (Netherlands)

    Tomà, T.; Courage, W.

    1996-01-01

    European Recommendations exist which provide calculation rules to determine the strength and stiffness of sandwich panels composed of two metal faces with a foam in between. In case of openings in such panels (e.g. for windows) an influence will appear with regard to the stiffness and loadbearing ca

  2. X-joints in composite sandwich panels

    NARCIS (Netherlands)

    Vredeveldt, A.W.; Janssen, G.Th.M.

    1998-01-01

    The small structural weight of fast large ships such as fast mono hulls or catamaran type of ships is of extreme importance to their success. One possible light weight structural solution is the sandwich panel with fibre reinforced laminates and a balsa, honeycomb or foam core. A severe obstacle for

  3. Organometallic half-sandwich iridium anticancer complexes

    NARCIS (Netherlands)

    Liu, Z.; Habtemariam, A.; Pizarro, A.M.; Fletcher, S.A.; Kisova, A.; Vrana, O.; Salassa, L.; Bruijnincx, P.C.A.|info:eu-repo/dai/nl/33799529X; Clarkson, G.J.; Brabec, V.; Sadler, Peter J.

    2011-01-01

    The low-spin 5d6 IrIII organometallic half-sandwich complexes [(η5-Cpx)Ir(XY)Cl]0/+, Cpx = Cp*, tetramethyl(phenyl)cyclopentadienyl (Cpxph), or tetramethyl(biphenyl)cyclopentadienyl (Cpxbiph), XY = 1,10-phenanthroline (4−6), 2,2′-bipyridine (7−9), ethylenediamine (10 and 11), or picolinate (12−14),

  4. Feedback Sandwiches Affect Perceptions but Not Performance

    Science.gov (United States)

    Parkes, Jay; Abercrombie, Sara; McCarty, Teresita

    2013-01-01

    The feedback sandwich technique-make positive comments; provide critique; end with positive comments-is commonly recommended to feedback givers despite scant evidence of its efficacy. These two studies (N = 20; N = 350) of written peer feedback with third-year medical students on clinical patient note-writing assignments indicate that students…

  5. X-joints in composite sandwich panels

    NARCIS (Netherlands)

    Vredeveldt, A.W.; Janssen, G.Th.M.

    1998-01-01

    The small structural weight of fast large ships such as fast mono hulls or catamaran type of ships is of extreme importance to their success. One possible light weight structural solution is the sandwich panel with fibre reinforced laminates and a balsa, honeycomb or foam core. A severe obstacle for

  6. Transferability of antibody pairs from ELISA to fiber optic surface plasmon resonance for infliximab detection

    Science.gov (United States)

    Van Stappen, Thomas; Lu, Jiadi; Bloemen, Maarten; Geukens, Nick; Spasic, Dragana; Delport, Filip; Verbiest, Thierry; Lammertyn, Jeroen; Gils, Ann

    2015-03-01

    Tumor necrosis factor (TNF)-alpha is a pleiotropic cytokine up-regulated in inflammatory bowel disease, rheumatoid arthritis and psoriasis. The introduction of anti-TNF drugs such as infliximab has revolutionized the treatment of these diseases. Recently, therapeutic drug monitoring (TDM) of infliximab has been introduced in clinical decision making to increase cost-efficiency. Nowadays, TDM is performed using radio-immunoassays, homogeneous mobility shift assays or ELISA. Unfortunately, these assays do not allow for in situ treatment optimization, because of the required sample transportation to centralized laboratories and the subsequent assay execution time. In this perspective, we evaluated the potential of fiber optic-surface plasmon resonance (FO-SPR). To achieve this goal, a panel of 55 monoclonal anti-infliximab antibodies (MA-IFX) was developed and characterized in-house, leading to the identification of nine different clusters. Based on this high diversity, 22 antibody pairs were selected and tested for their reactivity towards IFX, using one MA-IFX as capture and one MA-IFX for detection, in a sandwich type ELISA and FO-SPR. This study showed that the reactivity towards IFX of each antibody pair in ELISA is highly similar to its reactivity on FO-SPR, indicating that antibody pairs are easily transferable between both platforms. Given the fact that FO-SPR shows the potential for miniaturization and fast assay time, it can be considered a highly promising platform for on-site infliximab monitoring.

  7. Development of an ELISA for evaluation of swab recovery efficiencies of bovine serum albumin.

    Directory of Open Access Journals (Sweden)

    Nadja Sparding

    Full Text Available After a potential biological incident the sampling strategy and sample analysis are crucial for the outcome of the investigation and identification. In this study, we have developed a simple sandwich ELISA based on commercial components to quantify BSA (used as a surrogate for ricin with a detection range of 1.32-80 ng/mL. We used the ELISA to evaluate different protein swabbing procedures (swabbing techniques and after-swabbing treatments for two swab types: a cotton gauze swab and a flocked nylon swab. The optimal swabbing procedure for each swab type was used to obtain recovery efficiencies from different surface materials. The surface recoveries using the optimal swabbing procedure ranged from 0-60% and were significantly higher from nonporous surfaces compared to porous surfaces. In conclusion, this study presents a swabbing procedure evaluation and a simple BSA ELISA based on commercial components, which are easy to perform in a laboratory with basic facilities. The data indicate that different swabbing procedures were optimal for each of the tested swab types, and the particular swab preference depends on the surface material to be swabbed.

  8. Hydrogen storage property of sandwiched magnesium hydride nanoparticle thin film

    Energy Technology Data Exchange (ETDEWEB)

    Barcelo, Steven; Rogers, Matthew; Grigoropoulos, Costas P.; Mao, Samuel S. [Lawrence Berkeley National Laboratory and Department of Mechanical Engineering, University of California at Berkeley, Berkeley, CA 94720 (United States)

    2010-07-15

    Hydrogen sorption property of magnesium (Mg) in the form of sandwiched Pd/Mg/Pd films is investigated. Pulsed laser deposition method was applied to deposit the samples consisting of films of nanoparticles. The enthalpy of formation of MgH{sub 2} was found to be -68 kJ/mol H{sub 2} for films with nanoparticle size on the order of 50 nm, which is smaller than the value for bulk MgH{sub 2} and may be explained by the concept of excess volume. (author)

  9. Optimization of the curing process of a sandwich panel

    Science.gov (United States)

    Phyo Maung, Pyi; Tatarnikov, O.; Malysheva, G.

    2016-10-01

    This study presented finite element modelling and experimental measurements of temperatures during the autoclave curing of the T-50 aircraft wing sandwich panel. This panel consists of upper and lower carbon fibre based laminates and an aluminium foil honeycomb. The finite element modelling was performed using the Femap-Nastran product. During processing, the temperature at various points on the surface of the panel was measured using the thermocouples. The finite element method simulated the thermal conditions and determined the temperatures in the different parts of the panel for a full cycle of the curing process. A comparison of the calculated and experimental data shows that their difference does not exceed 6%.

  10. FINITE ELEMENT MODELING OF THIN CIRCULAR SANDWICH PLATES DEFLECTION

    Directory of Open Access Journals (Sweden)

    K. S. Kurachka

    2014-01-01

    Full Text Available A mathematical model of a thin circular sandwich plate being under the vertical load is proposed. The model employs the finite element method and takes advantage of an axisymmetric finite element that leads to the small dimension of the resulting stiffness matrix and sufficient accuracy for practical calculations. The analytical expressions for computing local stiffness matrices are found, which can significantly speed up the process of forming the global stiffness matrix and increase the accuracy of calculations. A software is under development and verification. The discrepancy between the results of the mathematical model and those of analytical formulas for homogeneous thin circularsandwich plates does not exceed 7%.

  11. Sandwich Hologram Interferometry For Determination Of Sacroiliac Joint Movements

    Science.gov (United States)

    Vukicevic, S.; Vinter, I.; Vukicevic, D.

    1983-12-01

    Investigations were carried out on embalmed and fresh specimens of human pelvisis with preserved lumbar spines, hip joints and all the ligaments. Specimens were tested under static vertical loading by pulsed laser interferometry. The deformations and behaviour of particular pelvic parts were interpreted by providing computer interferogram models. Results indicate rotation and tilting of the sacrum in the dorso-ventral direction and small but significant movements in the cranio-caudal direction. Sandwich holography proved to be the only applicable method when there is a combination of translation and tilt in the range of 200 μm to 1.5 mm.

  12. Optical and thermophysical parameters measurement using sandwich photodetectors

    Science.gov (United States)

    Turinov, Valery I.

    1993-12-01

    A method is reported for estimating thermophysical and optical parameters, in which a two- spectral band sandwich photodetector is used. The sensitivity ranges of such a sensor are 2 - 5 micrometers and 8 - 14 micrometers . On irradiating an object's surface by a short radiation pulse the photodetector collected signals U1, U2 and the rates of their changes (dU1/dt), (dU2/dt) are recorded. These pertain to the same area on the object's surface and belong to the two spectral ranges mentioned. The parameters of the object are evaluated by means of calculations with the values measured.

  13. Novel CagA ELISA exhibits enhanced sensitivity of Helicobacter pylori CagA antibody

    Science.gov (United States)

    Matsuo, Yuichi; Kido, Yasutoshi; Akada, Junko; Shiota, Seiji; Binh, Tran Thanh; Trang, Tran Thi Huyen; Dung, Ho D Q; Tung, Pham Huu; Tri, Tran Dinh; Thuan, Ngo P Minh; Tam, Le Quang; Nam, Bui Chi; Khien, Vu Van; Yamaoka, Yoshio

    2017-01-01

    AIM To develop a novel Helicobacter pylori (H. pylori) CagA antibody enzyme-linked immunosorbent assay (ELISA) suitable for detecting serum anti-CagA antibodies with high sensitivity. METHODS Recombinant East Asian-type CagA protein was purified and immobilized for ELISA. Serum samples from 217 Vietnamese individuals (110 H. pylori-infected and 107 uninfected individuals) were applied. Conventional ELISA from Western-type CagA and our East Asian-type CagA ELISA were evaluated by comparing 38 subjects with the Western-type genotype and 72 subjects with the East Asian-type cagA genotype. Histological scores of the gastric mucosa were determined using the updated Sydney System to examine the relationship with anti-CagA antibody titers. RESULTS Recombinant 70-100 kDa fragments were immobilized on the ELISA plate. In ROC analysis, the area under the curve of our East Asian-type CagA ELISA was comparable to that of conventional CagA ELISA. The sensitivity of the two ELISAs differed depending on the cagA genotype. The sensitivity of East Asian-type CagA ELISA was higher for subjects infected with East Asian-type cagA H. pylori (P < 0.001), and the sensitivity of the conventional CagA ELISA tended to be higher for subjects infected with Western cagA H. pylori (P = 0.056). The titer of anti-CagA antibody tended to correlate with monocyte infiltration scores (r = 0.25, P = 0.058) and was inversely correlated with H. pylori density (r = -0.26, P = 0.043). CONCLUSION The novel ELISA is useful to detect anti-CagA antibodies in East Asian countries, and the titer may be a marker for predicting chronic gastritis. PMID:28104980

  14. Equivalent material modelling of sandwich beams, evanescent solutions and damping investigations

    Science.gov (United States)

    de Rijk, Sophie; Nijman, Eugene

    2016-11-01

    A novel method for representing the transverse vibrations of sandwich beams as equivalent Timoshenko beams is developed. Special attention is given to damping modelling together with the evanescent parts of the solutions to assert applicability of the approach to any boundary conditions. Shear stiffness is evaluated based on current knowledge. The latter is then used to update the reference theory for vibrations in sandwich beams. Analytical case studies are presented to show the performance and limitations of the method and compared with experimental data.

  15. Quasi-circular orbits of conformal thin-sandwich puncture binary black holes

    CERN Document Server

    Hannam, M D

    2005-01-01

    I construct initial data for equal-mass irrotational binary black holes using the conformal thin-sandwich puncture (CTSP) approach. I locate quasi-circular orbits using the effective-potential method, and estimate the location of the innermost stable circular orbit (ISCO). The ISCO prediction is consistent with results for conformal thin-sandwich data produced using excision techniques. These results also show that the ISCOs predicted by the effective-potential and ADM-Komar mass-comparison methods agree for CTS data, just as they did for Bowen-York data.

  16. Degradation of shear stiffness of Nomex honeycomb sandwich panel in laser irradiation

    Science.gov (United States)

    Wang, Jiawei; Jiang, Houman; Wu, Lixiong; Zhu, Yongxiang; Wei, Chenghua; Ma, Zhiliang; Wang, Lijun

    2017-05-01

    Based on the overhanging beam three-point bending method, the experimental system was set up to measure the variety of shear stiffness of Nomex honeycomb sandwich panel in laser irradiation. The shear stiffness of the specimens under different laser power density was measured. The result shows that the thermal effect during the laser irradiation leads to the degradation of mechanical properties of Nomex honeycomb sandwich panel. High temperature rise rate in the specimen is another main reason for the shear stiffness degeneration. This research provides a reference for the degradation of mechanical properties of composite materials in laser irradiation and proposes a new method for the study of laser interaction with matter.

  17. Nonlinear vibration and buckling of circular sandwich plate under complex load

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    The nonlinear vibration fundamental equation of circular sandwich plate under uniformed load and circumjacent load and the loosely clamped boundary condition were established by von Karman plate theory, and then accordingly exact solution of static load and its numerical results were given. Based on time mode hypothesis and the variational method, the control equation of the space mode was derived, and then the amplitude frequency-load character relation of circular sandwich plate was obtained by the modified iteration method. Consequently the rule of the effect of the two kinds of load on the vibration character of the circular sandwich plate was investigated. When circumjacent load makes the lowest natural frequency zero, critical load is obtained.

  18. Study of sandwiched three-port transmission grating with a connection layer under normal incidence

    Science.gov (United States)

    Li, Hongtao; Wang, Bo

    2016-09-01

    A sandwiched three-port transmission grating with a connection layer under normal incidence is shown. The sandwiched dielectric grating can mainly diffract the identical energies in the ±1st orders and the zeroth diffraction order based on optimized grating parameters by employing rigorous coupled-wave analysis (RCWA) for TE and TM polarizations. On account of optimized parameters, efficiencies more than 32.5% in each diffraction order for two polarizations can be gained under normal incidence. A simplified modal method is used to describe the mechanism of beam propagation and analyze diffraction behavior clearly in the grating. Most importantly, diffraction efficiencies numerically obtained by using RCWA can be in agreement with the rough theoretical analysis on account of the simplified modal method. Therefore, the connection-layer-based sandwiched three-port grating is significant for the further practical manufacture.

  19. 夹层杯集菌离心涂片法检测抗酸杆菌及影响检出率的因素分析%Detection of Acid-fast Bacilli by Sandwich Cup Collecting Bacteria and Smear method and Analysis of Factors Affecting the Detection Rate

    Institute of Scientific and Technical Information of China (English)

    朱志斌; 张吉波; 雷鸣; 孙庆华

    2012-01-01

    Objective To evaluate the clinical value of sandwich cup collecting bacteria and smear method (sandwich cup method) in detecting acid- fast bacilli and to explore the factors affecting the detection rate of the acid - fast bacilli in sputum specimens. Methods Sputum specimens of 735 inpatients with confirmed diagnosis of tuberculosis were collected. Sputum specimens of each patient included morning sputum, night sputum and random sputum. Each sputum specimen was subjected to sandwich cup method and improved Lowenstein Jensen culture (L - J culture). The positive detection rates of acid - fast bacilli were compared between the two detection methods and among sputum specimens of different natures and collected at different time. Results The positive rates of the sandwich cup method and the L - J culture in 2,205 sputum specimens were 28.9 % and 32.1% respectively, which were significantly different(χ2 =54.744,P<0.001). Having the L-J culture as the standard method, the sensitivity, specificity, positive predictive value and negative predictive value of the sandwich cup method were 89.39 %, 99.53 %, 98.90 % and 95.21 %, respectively. The detection rates of acid - fast bacilli by the sandwich cup method in purulent sputum, bloody sputum, mucous sputum and saliva were 53. 3%,48.5%,9.9% and 3. 7%, respectively. TheL —J culture also had the highest detection rate in purulent sputum(54. 8%), followed by bloody sputum(51. 3%), mucous sputum (13.5%)and saliva (8.8%). For sputum samples collected at different time, the detection rates by both methods were the highest in the morning sputum. Conclusions Sandwich cup collecting bacteria and smear method is simple, time- saving, with high detection rate and more favorable to be standardized. It shows a good coincidence with the L- J culture. It can be used as a valuable detection method for acid- fast bacillus and can be popularized. Both methods have high detection rates in morning sputum, purulent sputum and bloody sputum

  20. HYBRID-SANDWICHED REINFORCEMENT WITH GEOSYNTHETICS

    Science.gov (United States)

    Yasuhara, Kazuya; Yamazaki, Shinji; Sakakibara, Tsutomu

    Advantageous aspects of sandwich-type reinforced earth structures combined with geosynthetics and sand mat are highlighted in this paper. Those aspects were elucidated by two kinds of laboratory tests : (1) large consolidation tests for improvement of hydraulic conductivity and (2) model footing tests on improvement of bearing capacity and deformation characteristics for reinforced earth structures, including both vertical permeability and horizontal transmissibility characteristics of geosynthetics results from both laboratory tests indicated the following: i) Hydraulic conductivity of geosynthetics used for this type of earth reinforcement can be maintained for a long period. Such conductivity sometimes disappears, particularly because of clogging when geosynthetics are adopted in embankment construction using fine-grained soils. This fact indicates that the sand mats which are laid above and beneath geosynthetics play a salient role in preventing clogging of geosynthetics that occurs by intrusion of fines from cohesive soils. ii) Sandwich-type reinforcement combined with geosynthetics and sand mats increases stability and decreases deformation of earth structures. In particular, the sandwich structure is effective for providing toughness, which has remained an important issue for reducing infrastructural maintenance and costs. In the later part of the paper, conventionally available stability analysis was carried out to propose the design procedure for reinforced earth structures and at the same time numerical analysis was also conducted to ensure the applicability of the hybrid-sandwiched earth reinforcement newly proposed in the current paper. Finally, based on the horizontal placement by means of HBS described in the current paper, the vertical drain procedure using the sandwich structures for accelerating consolidation and increasing stability of soft soils is also suggested for the future research and investigation.

  1. Pemendekan Waktu ELISA dalam Deteksi TMV

    Directory of Open Access Journals (Sweden)

    Susamto Somowiyarjo

    1996-12-01

    Full Text Available In order to support the program for the management of viral diseases on garlic, a rapid diagnosis procedure was developed by shortening the incubation period of the indirect ELISA. No significant differences of ELISA absorbance were observed when the antigen was incubated for 4 h, 2 h, and 30 min at 37ºC. The incubation period for the antibody and conjugate could he reduced from 4 h at 37ºC or 18 h at 4ºC to 30 min al 37ºC. The shortest period of incubation for the assay could be obtained when each incubation time for the antigen, antibody and conjugate was 30 min. The detection limit for the shortened ELISA was 10^-4 for the virus in crude extracts and 0,5 μg/ml for the purified preparation.

  2. Human toxocariosis: Seroprevalence in Lima inhabitans by ELISA technique

    OpenAIRE

    ESPINOZA, YRMA; Instituto de Medicina Tropical “Daniel A. Carrión”, Facultad de Medicina, UNMSM; Departamento Académico de Microbiología Médica Facultad de Medicina, UNMSM; Huapaya, Pedro; Instituto de Medicina Tropical “Daniel A. Carrión”, Universidad Nacional Mayor de San Marcos; Sevilla, Carlos; Departamento Académico de Microbiología Médica; Huiza, Alina; Instituto de Medicina Tropical “Daniel A. Carrión”, Universidad Nacional Mayor de San Marcos, Lima, Perú. Bióloga.; Jiménez, Susana; Departamento Académico de Microbiología Médica, Facultad de Medicina, Universidad Nacional Mayor de San Marcos. Lima, Perú.; Náquira, César; Instituto de Medicina Tropical “Daniel A. Carrión”, UNSMM; Instituto Nacional de Salud

    2013-01-01

    Objective: To estimate seroprevalence of human toxocariosis in Lima inhabitants. Design: Cross-sectional study, non aleatory selection. Material and methods: To people living in Lima city urban marginal communities, an interview and clinical examination were done and serum samples obtained to detect antibodies against Toxocara by ELISA technique. Stool samples were also obtained to check parasites causing cross reactions with serology. Results: From 553 persons examined 23,3% were reactive. T...