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Sample records for enzyme immunoassay eia

  1. Evaluation and Comparison of Enzyme Immunoassay (Eia and Acid Fast Staining with Confirmation by Immunofluorescent Antibody Assay for Detection of Cryptosporidium Species in Infants and Young Children.

    Directory of Open Access Journals (Sweden)

    D Dorostcar Moghaddam

    2005-01-01

    Full Text Available Introduction: Cryptosporidiosis is prevalent world wide, causing a variety of problems ranging from acute, self-limiting diarrhea to fatal cases in immunocompromised persons, particulary those with acquired immunodeficiency (AIDS. Diagnosis of Cryptosporidium is made by identification of oocysts in stool specimens. The detection is most commonly made by the acid-fast staining method followed by microscopic examination which has low specificity and sensitivity. Material and Methods: In the present study, we evaluated diagnostic utility of a commercially available enzyme immunoassay (EIA, which detects Cryptosporidium-Specific antigen (CSA in 204 unprocessed stool specimens obtained from patients less than 3 years of age. Results: When compared with the routine screening procedure applied in this field study (screening by acid-fast staining and microscopy after concentration of positive results by IFA, both sensitivity and specificity were 98%. Of the 139 specimens negative by microscopy, 13 (9.3% were positive by EIA, 11 of which were confirmed by inhibition with antibody to Cryptosporidia-specific antigen. Conclusion: The EIA is an important tool for identifying Cryptosporidium in fecal specimens in field studies since it is sensitive, specific, simple to use and unaffected by the presence of a preservative.

  2. Estimating False-Recent Classification for the Limiting-Antigen Avidity EIA and BED-Capture Enzyme Immunoassay in Vietnam: Implications for HIV-1 Incidence Estimates.

    Science.gov (United States)

    Shah, Neha S; Duong, Yen T; Le, Linh-Vi; Tuan, Nguyen Anh; Parekh, Bharat S; Ha, Hoang Thi Thanh; Pham, Quang Duy; Cuc, Cao Thi Thu; Dobbs, Trudy; Tram, Tran Hong; Lien, Truong Thi Xuan; Wagar, Nick; Yang, Chunfu; Martin, Amy; Wolfe, Mitchell; Hien, Nguyen Tran; Kim, Andrea A

    2017-06-01

    Laboratory tests that can distinguish recent from long-term HIV infection are used to estimate HIV incidence in a population, but can potentially misclassify a proportion of long-term HIV infections as recent. Correct application of an assay requires determination of the proportion false recents (PFRs) as part of the assay characterization and for calculating HIV incidence in a local population using a HIV incidence assay. From April 2009 to December 2010, blood specimens were collected from HIV-infected individuals attending nine outpatient clinics (OPCs) in Vietnam (four from northern and five from southern Vietnam). Participants were living with HIV for ≥1 year and reported no antiretroviral (ARV) drug treatment. Basic demographic data and clinical information were collected. Specimens were tested with the BED capture enzyme immunoassay (BED-CEIA) and the Limiting-antigen (LAg)-Avidity EIA. PFR was estimated by dividing the number of specimens classified as recent by the total number of specimens; 95% confidence intervals (CI) were calculated. Specimens that tested recent had viral load testing performed. Among 1,813 specimens (north, n = 942 and south, n = 871), the LAg-Avidity EIA PFR was 1.7% (CI: 1.2-2.4) and differed by region [north 2.7% (CI: 1.8-3.9) versus south 0.7% (CI: 0.3-1.5); p = .002]. The BED-CEIA PFR was 2.3% (CI: 1.7-3.0) and varied by region [north 3.4% (CI: 2.4-4.7) versus south 1.0% (CI: 0.5-1.2), p < .001]. Excluding specimens with an undetectable VL, the LAg-Avidity EIA PFR was 1.2% (CI: 0.8-1.9) and the BED-CEIA PFR was 1.7% (CI: 1.2-2.4). The LAg-Avidity EIA PFR was lower than the BED-CEIA PFR. After excluding specimens with an undetectable VL, the PFR for both assays was similar. A low PFR should facilitate the implementation of the LAg-Avidity EIA for cross-sectional incidence estimates in Vietnam.

  3. Enzyme immunoassay

    DEFF Research Database (Denmark)

    Feldt-Rasmussen, B; Dinesen, B; Deckert, M

    1985-01-01

    An enzyme linked immunoadsorbent assay for urinary albumin using commercially available reagents is described. The assay range is 2.5-120 micrograms/l. When samples are analysed in two standard dilutions, the assayable albumin concentration range is 2.5-240 mg/l, covering the clinical range from...

  4. Detection of poliovirus antigen by enzyme immunoassay.

    OpenAIRE

    Ukkonen, P; Huovilainen, A; Hovi, T

    1986-01-01

    A solid-phase enzyme immunoassay (EIA) was developed for the detection of poliovirus antigen. Rabbit and guinea pig antisera for the assay were raised against purified poliovirus type 3/Fin (strain 3/Fin/K) isolated from a fecal specimen from a meningitis patient during an outbreak of poliomyelitis in Finland in 1984. The EIA was highly specific for poliovirus type 3, and it was about 30 times more sensitive for strain 3/Fin/K than for strain 3/Saukett used in the inactivated poliovirus vacci...

  5. Comparison of an enzyme-immunoassay with a radio-immunoassay ...

    African Journals Online (AJOL)

    1990-07-21

    HBs, ... (RIA) and enzyme-immunoassay (EIA) method for detecting the hepatitis markers anti-HBs, anti-HBc and ... radioactive anti-HBc in the serum competes with human anti-. HBc 1251 for binding sites on beads coated with ...

  6. Development, evaluation, and laboratory validation of immunoassays for the diagnosis of equine infectious anemia (EIA) using recombinant protein produced from a synthetic p26 gene of EIA virus.

    Science.gov (United States)

    Singha, Harisankar; Goyal, Sachin K; Malik, Praveen; Khurana, Sandip K; Singh, Raj K

    2013-12-01

    Equine infectious anemia (EIA)-a retroviral disease caused by equine infectious anemia virus (EIAV)-is a chronic, debilitating disease of horses, mules, and donkeys. EIAV infection has been reported worldwide and is recognized as pathogen of significant economic importance to the horse industry. This disease falls under regulatory control program in many countries including India. Control of EIA is based on identification of inapparent carriers by detection of antibodies to EIAV in serologic tests and "Stamping Out" policy. The current internationally accepted test for diagnosis of EIA is the agar gel immune-diffusion test (AGID), which detects antibodies to the major gag gene (p26) product. The objective of this study was to develop recombinant p26 based in-house immunoassays [enzyme linked immunosorbent assays (ELISA), and AGID] for EIA diagnosis. The synthetic p26 gene of EIAV was expressed in Escherichia coli and diagnostic potential of recombinant p26 protein were evaluated in ELISA and AGID on 7,150 and 1,200 equine serum samples, respectively, and compared with commercial standard AGID kit. The relative sensitivity and specificity of the newly developed ELISA were 100 and 98.6 %, respectively. Whereas, relative sensitivity and specificity of the newly developed AGID were in complete agreement in respect to commercial AGID kit. Here, we have reported the validation of an ELISA and AGID on large number of equine serum samples using recombinant p26 protein produced from synthetic gene which does not require handling of pathogenic EIAV. Since the indigenously developed reagents would be economical than commercial diagnostic kit, the rp26 based-immunoassays could be adopted for the sero-diagnosis and control of EIA in India.

  7. Development of a highly specific and sensitive rubella immunoglobulin M antibody capture enzyme immunoassay that uses enzyme-labeled antigen.

    OpenAIRE

    Seppänen, H

    1990-01-01

    An enzyme immunoassay (EIA) for serum immunoglobulin M (IgM) antibodies to rubella virus based on enzyme labeling of viral antigen was developed. The sensitivity of the EIA for the detection of recent rubella virus infection was evaluated by using 115 rubella-IgM-antibody-positive serum specimens, which were confirmed as positive by Rubazyme M (Abbott Diagnostics). In addition, 12 individuals, 2 of whom were exposed to rubella through vaccination and 10 of whom were exposed through natural in...

  8. Thermophilic Campylobacter spp. in turkey samples: evaluation of two automated enzyme immunoassays and conventional microbiological techniques

    DEFF Research Database (Denmark)

    Borck, Birgitte; Stryhn, H.; Ersboll, A.K.

    2002-01-01

    Aims: To determine the sensitivity and specificity of two automated enzyme immunoassays (EIA), EiaFoss and Minividas, and a conventional microbiological culture technique for detecting thermophilic Campylobacter spp. in turkey samples. Methods and Results: A total of 286 samples (faecal, meat......, neckskin and environmental samples) were collected over a period of 4 months at a turkey slaughterhouse and meat-cutting plant in Denmark. Faecal and environmental samples were tested by the conventional culture method and by the two EIAs, whereas meat and neckskin samples were tested by the two EIAs only...

  9. Estrogen receptor determination in endometrial carcinoma: ligand binding assay versus enzyme immunoassay

    DEFF Research Database (Denmark)

    Nyholm, H C; Nielsen, Anette Lynge; Lyndrup, J

    1995-01-01

    We compared concentrations of cytosolic estrogen receptors (ERc) measured in 35 postmenopausal endometrial carcinomas by ligand binding method (LBA) (dextran-coated charcoal assay) and enzyme immunoassay (EIA). Correlations between ERc, nuclear estrogen receptors (ERn) determined by EIA, and cyto......We compared concentrations of cytosolic estrogen receptors (ERc) measured in 35 postmenopausal endometrial carcinomas by ligand binding method (LBA) (dextran-coated charcoal assay) and enzyme immunoassay (EIA). Correlations between ERc, nuclear estrogen receptors (ERn) determined by EIA......, and cytosolic progesterone receptors (PR) measured by LBA were also studied. While ERc concentrations determined by LBA and EIA were highly correlated (r: 0.94), ERc values detected by LBA were approximately twice those found by EIA (median values of ERc: 155 vs. 64 fmol/mg cytosol protein, DCC vs. EIA......). The percentages of ERc positive tumors were 89% by LBA and 77% by EIA. The median fraction of total ER present as ERn was 63%. PR levels correlated positively with ERn concentrations (r: 0.73). We explore possible reasons why greater concentrations of ERc are determined by estradiol binding than by the ER-EIA kit...

  10. Nitrocellulose membrane-based enzyme-linked immunoassay for dengue serotype-1 IgM detection

    International Nuclear Information System (INIS)

    Leon, S.; Guevara, C.; Chunga, A.

    1999-01-01

    To evaluate the sensitivity and specifity of a nitrocellulose membrane-based immunoassay for dengue IgM, with respect to capture enzyme immunoassay, for the diagnosis of dengue virus infection. 101 serum samples were processed and divided into 2 groups: 53 from dengue serotype 1 (DEN1) infected patients, and 48 from healthy subjects. Both groups were tested with a nitrocellulose membrane-based IgM capture enzyme immunoassay (NMB-EIA) and also with an ELISA as referential pattern. NMB-EIA testing detected IgM anti-DEN1 in 94,34% of samples from infected patients, and in 14,58% of control samples, whereas ELISA fails to report false positive or false negative results: NMB-EIA appears to be a good alternative for dengue infection diagnosis. (authors)

  11. Comparison of an enzyme-immunoassay with a radio-immunoassay ...

    African Journals Online (AJOL)

    immunoassay (EIA) method for detecting the hepatitis markers anti-HBs, anti-HBc and HBsAg. The results indicated that the RIA and EIA were comparable for the HBsAg marker but that the RIA test was more sensit!ve for anti-HBs and more ...

  12. Comparative digoxin determination in serum by means of a radioimmuno- and an enzyme immunoassay

    International Nuclear Information System (INIS)

    Reinhard, M.

    1981-01-01

    Two immunologic measuring methods for the quantitative digoxin determination in serum were compared. One method bases on the principle of radioisotope dilution, the second one on the principle of enzyme inhibition. The radioimmunoassay served as reference method. The limit of detection for RIA is 0.23 ng/ml, for EIA 0.40 ng/ml. For both methods the measuring range extends up to approx. 5.5 ng/ml. The degree of precision in series is 8.2% for RIA, 10.8% for EIA. Day-to-day precision is 4.4% for RIA, 15.2% for EIA. On comparison, the 59 serum samples of patients who received digoxin did not show any systematic difference. The results obtained can be transformed by means of the equations Csub(EIA) = 0.041 ng/ml + 0.936 Csub(RIA). In pathologic sera, however, there are significant differences disfavoring EIA, because due to high color concentrations or turbidities these sera do not permit any or any exact extinction measurements. The enzyme immunoassay should not be used with such sera. With regard to practicability the EIA corresponds more or less to RIA. The EIA can essentially be economized by using semi-microcuvettes and applying only the half of the recommended enzyme and antibody volume. (orig.) [de

  13. Determination of fenoterol and ractopamine in urine by enzyme immunoassay.

    Science.gov (United States)

    Haasnoot, W; Stouten, P; Lommen, A; Cazemier, G; Hooijerink, D; Schilt, R

    1994-12-01

    Fenoterol and ractopamine are phenethanolamines with beta-adrenergic agonist activity. An enzyme immunoassay (EIA) for these compounds was developed using antibodies raised in a New Zealand white rabbit against fenoterol-bovine serum albumin and fenoterol coupled to horseradish peroxidase (HRP). The calibration graphs of fenoterol and ractopamine showed linearity over the concentration ranges 0.1-5 and 0.2-25 ng ml-1, respectively. Isoxsuprine showed a cross-reactivity of 0.7% while the cross-reactivity of other beta-agonists was combination with isobutanol extraction, the mean concentration was eight times higher than that obtained when using extraction only. Gas chromatography-mass spectrometry (GC-MS) analysis confirmed the presence of fenoterol in most of these samples. However, probably owing to the absence of a proper GC-MS internal standard, the correlation between GC-MS and EIA concentrations was low (r = 0.7976). In general, the concentrations found by the EIA are much higher than those found by GC-MS, which might be caused by the presence of metabolites detected with the EIA. Fenoterol is excreted in urine mostly (about 85%) as glucuronidated-sulfated conjugate. The antibodies partly recognize the conjugated fenoterol, which makes it possible to use a direct screening assay. In blank calf urine the detection limits, mean background +3 times the standard deviation, are 1.3 (fenoterol) and 2.6 ng ml-1 (ractopamine). In bovine urine, however, owing to matrix effects, the detection limits are 20 times higher.

  14. Comparison of clinical performance of antigen basedenzyme immunoassay (EIA and major outer membrane protein (MOMP-PCR for detection of genital Chlamydia trachomatis infection

    Directory of Open Access Journals (Sweden)

    Mahmoud Nateghi Rostami

    2016-06-01

    Full Text Available Background: Chlamydia trachomatis is the most common sexually transmitted bacterial pathogen worldwide. Early detection and treatment of C.trachomatis genital infection prevent serious reproductive complications. Objective: Performances of enzyme immunoassay (EIA and major outer membrane protein (MOMP-polymerase chain reaction (PCR for diagnosis of genital C.trachomatis infection in women were compared. Materials and Methods: In this cross sectional study a total of 518 women volunteers were included (33.67±8.3 yrs who had been referred to Gynecology clinics of Qom province, Iran, were included. Endocervical swab specimens were collected to detect lipopolysaccharide (LPS antigen in EIA and to amplify MOMP gene of C.trachomatis in PCR. Results were confirmed using ompI nested-PCR. Sensitivity, specificity, positive (PPV and negative predictive values (NPV were calculated for performance of the tests. Odds ratios were determined using binary logistic regression analysis. Results: In total, 37 (7.14% cases were positive by EIA and/or MOMP-PCR. All discrepant results were confirmed by nested-PCR. Sensitivity, specificity, PPV and NPV values of EIA were 59.46%, 100%, 100% and 96.98%, and those of MOMPPCR were 97.30%, 100%, 100%, 99.79%, respectively. Reproductive complications including 2.7% ectopic pregnancy, 5.4% stillbirth, 5.4% infertility, and 10.8% PROM were recorded. The risk of developing chlamydiosis was increased 4.8-fold in volunteers with cervicitis (p<0.05; OR 4.80; 95% CI 1.25-18.48. Conclusion: C.trachomatis infection should be regarded in women of reproductive ages especially those with cervicitis. Primary screening of women by using the low cost antigen-EIA is recommended; however, due to the low sensitivity of Ag-EIA, verification of the negative results by a DNA amplification method is needed.

  15. Comparative studies on the determination of alphafetoprotein by enzyme immunoassay and by radioimmunoassay

    International Nuclear Information System (INIS)

    Haller, G.; Linneke, P.; Voss, P.; Jeske, W.

    1987-01-01

    Alphafetoprotein (AFP) was determined in serum of pregnant women in the tenth till sixteenth week of pregnancy by means of two enzyme immunoassays (Enzymun-Test AFP, Boehringer Mannheim, FRG and AFP EIA 'Dessau' 1000, Research Institute for Vaccine Dessau, GDR) and a radioimmunoassay (Radioimmunoassay Kit, AFP-PR, CIS, France). Parallel determinations in sera of 438 patients, who had come to surveillance for the first consultation were estimated. A comparison between the methods showed a good correlation. (author)

  16. Evaluation of an enzyme immunoassay for verotoxin detection in Escherichia coli.

    Science.gov (United States)

    Frias, C; Majò, M; Margall, N; Llobet, T; Mirelis, B; Prats, G

    1996-09-01

    Verotoxin-producing Escherichia coli strains (VTEC) cause hemorrhagic colitis and hemolytic-uremic syndrome in humans. Laboratory diagnosis by conventional methods is slow and cumbersome. The results of a new rapid enzyme immunoassay (EIA Premier EHEC) for verotoxin detection both in isolated strains and in clinical samples are presented, and they are compared with cell culture (CC) and polymerase chain reaction (PCR) techniques. Fifty-four strains have been analyzed by both EIA and PCR, and 33 by all three methods. The kit has also been evaluated for experimentally infected stool samples directly and after their enrichment on MacConkey broth. Nineteen, out of the 54 strains, were positive by EIA and 20 by PCR. The results of the 33 strains evaluated by the three techniques were coincident with one exception. The latter was uninterpretable by CC, negative by EIA and positive by PCR. The sensitivity of the kit for experimentally infected stool samples was approximately 5 x 10(7) bacteria/ml in the direct test, and 5 x 10(4) bacteria/ml after broth enrichment. EIA sensitivity and specificity were similar to those of CC and PCR. The diagnostic times were 18h for EIA, 3 days for PCR and 5 days for CC. Sensitivity, rapidity and ease of performance make this technique especially valuable for clinical diagnosis.

  17. Comparison of Salivary and Serum Enzyme Immunoassays for the Diagnosis of Helicobacter pylori Infection

    Directory of Open Access Journals (Sweden)

    John M Embil

    1998-01-01

    Full Text Available Infection with Helicobacter pylori has been established as an important risk factor for the development of peptic ulcer disease, gastritis and gastric cancer. The diagnosis of H pylori infection can be established by invasive or noninvasive techniques. Two noninvasive enzyme immunoassays (EIAs for antibody detection – HeliSal and Pylori Stat – were compared with histology. Both assays detect immunoglobulin (Ig G directed against purified H pylori antigen. The test populations consisted of 104 consecutive patients scheduled for upper gastrointestinal endoscopy. Of these patients, 97 (93% had symptoms compatible with peptic ulcer disease. Saliva and serum were collected simultaneously at the time of endoscopy. Salivary EIA had a sensitivity of 66%, specificity of 67%, positive predictive value of 67% and negative predictive value of 66% compared with the serum EIA, where the results were 98%, 48%, 64% and 96%, respectively. Although the salivary EIA is an appealing noninvasive test, it was not a sensitive and specific assay. The serum EIA also lacked specificity, but was highly sensitive with a good negative predictive value. Although a negative serum EIA rules out H pylori infection, a positive result must be interpreted in the clinical context and confirmed with a more specific measure.

  18. Detection of antinuclear antibodies by solid-phase immunoassays and immunofluorescence analysis

    DEFF Research Database (Denmark)

    Fenger, Mogens; Wiik, Allan; Høier-Madsen, Mimi

    2004-01-01

    BACKGROUND: Antinuclear antibodies (ANAs) are associated with several inflammatory rheumatic diseases. The aim of the present work was to evaluate enzyme immunoassays (EIAs) and compare them with classic immunofluorescent analysis (IFA) for the detection of ANA. METHODS: Seven enzyme immunoassays...

  19. Failure of human immunodeficiency virus enzyme immunoassay to rule out infection among polymerase chain reaction-negative Vietnamese infants at 12 months of age.

    Science.gov (United States)

    Sohn, Annette H; Thanh, Tran Chi; Thinh, Le Quoc; Khanh, Truong Huu; Thu, Huynh Khanh; Giang, Le Truong; Lien, Truong Xuan

    2009-04-01

    : Previous studies have demonstrated that >90% of HIV-uninfected infants serorevert, as seen in the results of enzyme immunoassay (EIA) testing by 12 months of age, making it feasible to confirm or rule out infection. We assessed the reliability of EIA in a cohort of Vietnamese infants. : HIV-exposed, uninfected infants enrolled in a parent diagnostic and monitoring study from February 2005 through August 2006 were eligible for inclusion in a prospective cohort study of HIV-EIA performance. Testing using 2 standard assays (Genscreen HIV 1/2 version 2, Bio-Rad; Murex 1.2.0, Murex Biotech) was initiated at 12 months of age. Infants were categorized as EIA-negative (seroreverted; negative Genscreen), EIA-indeterminate (positive Genscreen, negative Murex), or EIA-positive (Genscreen and Murex positive). : Of 273 infants included in the study, 59 (22%) were EIA-negative at 12 months, 131 (48%) were indeterminate, and 83 (30%) were EIA-positive; specificity 21.6 (95% confidence interval: 16.6, 26.3). Infants with positive EIAs at 12 months were 74% more likely than EIA-indeterminate infants to test indeterminate or positive at 18 months (risk ratio, 1.74, 95% confidence interval: 1.15, 2.64; P = 0.03). : Expectations regarding infant seroreversion by standard EIAs should be reassessed to reflect potential cross-regional differences in their performance.

  20. Detection of anti-nuclear antibody by immunofluorescence assay and enzyme immunoassay in childhood systemic lupus erythematosus: experience from Bangladesh.

    Science.gov (United States)

    Dipti, Tanjeem Rabika; Azam, Mohammad Shaiful; Sattar, Mohammad Humayun; Rahman, Shahana Akhter

    2012-02-01

    Systemic lupus erythematosus (SLE) is a multisystem, chronic but often episodic, autoimmune disease that is characterized by the presence of antinuclear antibodies (ANA). The criteria set by American College of Rheumatology are widely used for diagnosis of SLE. Elevation of ANA titer is the most sensitive of the ACR criteria. There are different methods for detection of ANA. Indirect immunofluorescence (ANA-IFA) and enzyme immunoassay (ANA-EIA) are commonly used methods. The sensitivity of ANA-IFA using HEp-2 cell substrate is 90-100% in systemic rheumatic diseases. In Bangladesh most of the laboratories use ANA-EIA for detection of ANA. As the sensitivity of ANA-EIA is lower than ANA-IFA it might be that we are missing many cases of ANA positivity in childhood SLE cases. To detect ANA by immunofluorescence assay using HEp-2 cell substrate and enzyme immunoassay in childhood SLE and to compare the diagnostic performance of these methods. This is a cross-sectional analytical study. A total of 40 patients were enrolled. Among them 20 were childhood SLE cases. Another 20 patients of childhood rheumatic diseases other than SLE were taken as the disease control group. In childhood SLE cases, 100% were ANA-positive by IFA and 55% were ANA positive by EIA. The sensitivity of ANA-IFA was 100%. In contrast, sensitivity of ANA-EIA was 55%.   ANA-IFA is superior to ANA-EIA for detection of ANA in childhood SLE patients. ANA-IFA should be the primary screening test for children with clinical features suggestive of SLE. © 2011 The Authors. International Journal of Rheumatic Diseases © 2011 Asia Pacific League of Associations for Rheumatology and Blackwell Publishing Asia Pty Ltd.

  1. Rapid detection of fungal alpha-amylase in the work environment with a lateral flow immunoassay

    NARCIS (Netherlands)

    Bogdanovic, J.; Koets, M.; Sander, I.; Wouters, I.; Meijster, T.; Heederik, D.J.J.; Amerongen, van A.; Doekes, G.

    2006-01-01

    Background Occupational allergen exposure assessment usually requires airborne dust sampling at the worksite followed by dust extraction and enzyme immunoassay (EIA) analysis at the laboratory. Use of semiquantitative lateral flow immunoassays (LFIAs) may allow a more rapid detection procedure with

  2. A competitive enzyme immunoassay for the quantitative detection of cocaine from banknotes and latent fingermarks.

    Science.gov (United States)

    van der Heide, Susan; Garcia Calavia, Paula; Hardwick, Sheila; Hudson, Simon; Wolff, Kim; Russell, David A

    2015-05-01

    A sensitive and versatile competitive enzyme immunoassay (cEIA) has been developed for the quantitative detection of cocaine in complex forensic samples. Polyclonal anti-cocaine antibody was purified from serum and deposited onto microtiter plates. The concentration of the cocaine antibody adsorbed onto the plates, and the dilution of the cocaine-HRP hapten were both studied to achieve an optimised immunoassay. The method was successfully used to quantify cocaine in extracts taken from both paper currency and latent fingermarks. The limit of detection (LOD) of 0.162ngmL(-1) achieved with the assay compares favourably to that of conventional chromatography-mass spectroscopy techniques, with an appropriate sensitivity for the quantification of cocaine at the low concentrations present in some forensic samples. The cEIA was directly compared to LC-MS for the analysis of ten UK banknote samples. The results obtained from both techniques were statistically similar, suggesting that the immunoassay was unaffected by cross-reactivity with potentially interfering compounds. The cEIA was used also for the detection of cocaine in extracts from latent fingermarks. The results obtained were compared to the cocaine concentrations detected in oral fluid sampled from the same individual. Using the cEIA, we have shown, for the first time, that endogeneously excreted cocaine can be detected and quantified from a single latent fingermark. Additionally, it has been shown that the presence of cocaine, at similar concentrations, in more than one latent fingermark from the same individual can be linked with those concentrations found in oral fluid. These results show that detection of drugs in latent fingermarks could directly indicate whether an individual has consumed the drug. The specificity and feasibility of measuring low concentrations of cocaine in complex forensic samples demonstrate the effectiveness and robustness of the assay. The immunoassay presents a simple and cost

  3. Development of an enzyme immunoassay for poliovirus antigens

    Directory of Open Access Journals (Sweden)

    Newton Hashimoto

    2007-01-01

    Full Text Available An indirect solid-phase enzyme immunoassay (EIA was developed for the detection of poliovirus antigen. Virus antigen was obtained in LLC-MK2 cell cultures and used to prepare antibodies in rabbit and guinea pig. Antibodies were evaluated by double immunodiffusion and neutralization test. Optimal concentrations of guinea pig and rabbit immunoglobulins were determined by checkerboard titration. Microtitre plates were coated with 15.0 µg/ml guinea pig anti-polio immunoglobulin and rabbit anti-polio immunoglobulin at the concentration of 7.94 µg/ml was used as detecting antibody. The standard curve with eight different antigen concentrations in eight replicates resulted in a coefficient of variation (CV between 2.1% to 7.8%. The dose-response relationship was determined by simple linear regression with a coefficient of correlation (R² equal to 96.4%. The assay detected a minimum of 2.3 µg/ml poliovirus antigen.O trabalho apresenta o desenvolvimento de um ensaio imunoenzimático indireto para a detecção de antígeno de poliovírus. O antígeno viral foi obtido em cultura de células LLC-MK2 e usado para imunização de coelho e cobaia. Os soros hiperimunes foram avaliados por imunodifusão dupla e teste de neutralização. Após padronização, o soro de captura, produzido em cobaia, foi usado na concentração protéica de 15.0 µg/ml para sensibilizar microplacas de poliestireno e o soro de coelho (detector foi usado na concentração de 7.94 µg/ml. A curva padrão resultante da utilização de oito diferentes concentrações do antígeno padrão definiu um coeficiente de variação de 2.1% a 7.8%. A relação dose-resposta foi determinada por regressão linear simples com o estabelecimento do coeficiente de correlação (R² igual a 96.4%. O ensaio possibilitou a detecção mínima de 2.3 µg/ml de antígeno de poliovírus.

  4. An enzyme immunoassay for detection of Japanese encephalitis ...

    Indian Academy of Sciences (India)

    Japanese encephalitis virus (JEV) induces human peripheral blood monocytes to secrete a chemotactic cytokine [human macrophage-derived factor (hMDF)] which causes chemotaxis of neutrophils. The only known assay for hMDF cannot quantify its level in samples, so an enzyme immunoassay has been standardized for ...

  5. An enzyme immunoassay for detection of Japanese encephalitis ...

    Indian Academy of Sciences (India)

    Enzyme immunoassay; human macrophage-derived factor; Japanese encephalitis virus. Abbreviations used: ... macrophage-derived factor; HBSS, Hank's balanced salt solution; CSF, cerebrospinal fluid; MEM, minimum essential medium; FCS ..... much attention as pathogenic or marker substance in various diseases (Ida ...

  6. Enzyme immunoassay for progesterone and estradiol. A study of factors influencing sensitivity

    International Nuclear Information System (INIS)

    Joyce, B.G.; Read, G.F.; Riad-Fahmy, D.

    1978-01-01

    Enzyme immunoassays (EIA) for progesterone and estradiol have been developed at the Tenovus Institute using horseradish peroxidase conjugates and a solid-phase system for separation of free and bound steroid. The sensitivity of these assays was inadequate and precluded their use for the assay of low-titre samples. Attempts to increase the sensitivity of the assay by reducing the affinity of the conjugate for the antiserum by altering the bridge groups were unsuccessful. Decrease in the molar incorporation of the steroid into the enzyme conjugate effected a significant improvement, increasing the sensitivity in the progesterone assay by 50% and in that of estradiol by 70%. Initial attempts to replace the solid-phase separation technique by a second antibody procedure based on a sheep, anti-rabbit IgG serum were of limited success. Increased sensitivity in the progesterone radioimmunoassay (RIA) was achieved but severe problems with high non-specific binding remained. The apparent specific binding in the estradiol assay did not exceed 30% and a viable system could not therefore be established. An improved second antibody produced in guinea-pigs to a wider spectrum of immunoglobulins, when used in the EIA of progesterone, increased sensitivity 50-fold compared with the solid-phase assay. The sensitivity of the estradiol assay was increased 8-fold compared with the solid-phase system, and is now comparable with that of the RIA (2.0pg/0.7pg). The significant improvement in sensitivity in EIA associated with the use of double antibody separation procedures, and the importance of tailoring the affinity of the second antibody to the immunoglobulin profile of the first antibody are emphasized. (author)

  7. Detection of Campylobacter in Stool and Determination of Significance by Culture, Enzyme Immunoassay, and PCR in Developing Countries

    Science.gov (United States)

    Platts-Mills, James A.; Liu, Jie; Gratz, Jean; Mduma, Esto; Amour, Caroline; Swai, Ndealilia; Taniuchi, Mami; Begum, Sharmin; Peñataro Yori, Pablo; Tilley, Drake H.; Lee, Gwenyth; Shen, Zeli; Whary, Mark T.; Fox, James G.; McGrath, Monica; Kosek, Margaret; Haque, Rashidul

    2014-01-01

    Campylobacter is a common bacterial enteropathogen that can be detected in stool by culture, enzyme immunoassay (EIA), or PCR. We compared culture for C. jejuni/C. coli, EIA (ProSpecT), and duplex PCR to distinguish Campylobacter jejuni/C. coli and non-jejuni/coli Campylobacter on 432 diarrheal and matched control stool samples from infants in a multisite longitudinal study of enteric infections in Tanzania, Bangladesh, and Peru. The sensitivity and specificity of culture were 8.5% and 97.6%, respectively, compared with the results of EIA and 8.7% and 98.0%, respectively, compared with the results of PCR for C. jejuni/C. coli. Most (71.6%) EIA-positive samples were positive by PCR for C. jejuni/C. coli, but 27.6% were positive for non-jejuni/coli Campylobacter species. Sequencing of 16S rRNA from 53 of these non-jejuni/coli Campylobacter samples showed that it most closely matched the 16S rRNA of C. hyointestinalis subsp. lawsonii (56%), C. troglodytis (33%), C. upsaliensis (7.7%), and C. jejuni/C. coli (2.6%). Campylobacter-negative stool spiked with each of the above-mentioned Campylobacter species revealed reactivity with EIA. PCR detection of Campylobacter species was strongly associated with diarrhea in Peru (odds ratio [OR] = 3.66, P < 0.001) but not in Tanzania (OR = 1.56, P = 0.24) or Bangladesh (OR = 1.13, P = 0.75). According to PCR, Campylobacter jejuni/C. coli infections represented less than half of all infections with Campylobacter species. In sum, in infants in developing country settings, the ProSpecT EIA and PCR for Campylobacter reveal extremely high rates of positivity. We propose the use of PCR because it retains high sensitivity, can ascertain burden, and can distinguish between Campylobacter infections at the species level. PMID:24452175

  8. Use of a nested PCR-enzyme immunoassay with an internal control to detect Chlamydophila psittaci in turkeys

    Directory of Open Access Journals (Sweden)

    Goddeeris Bruno M

    2005-09-01

    Full Text Available Abstract Background Laboratory diagnosis of Chlamydophila psittaci, an important turkey respiratory pathogen, is difficult. To facilitate the diagnosis, a nested PCR-enzyme immunoassay (PCR-EIA was developed to detect the Cp. psittaci outer membrane protein A (ompA gene in pharyngeal swabs. Methods The fluorescein-biotin labelled PCR products were immobilized on streptavidin-coated microtiter plates and detected with anti-fluorescein peroxidase conjugate and a colorimetric substrate. An internal inhibition control was included to rule out the presence of inhibitors of DNA amplification. The diagnostic value of the ompA nested PCR-EIA in comparison to cell culture and a 16S-rRNA based nested PCR was assessed in pharyngeal turkey swabs from 10 different farms experiencing respiratory disease. Results The sensitivity of the nested PCR-EIA was established at 0.1 infection forming units (IFU. Specificity was 100%. The ompA nested PCR-EIA was more sensitive than the 16S-rRNA based nested PCR and isolation, revealing 105 out of 200 (52.5% positives against 13 and 74 for the latter two tests, respectively. Twenty-nine (23.8% out of 122 ompA PCR-EIA negatives showed the presence of inhibitors of DNA amplification, although 27 of them became positive after diluting (1/10 the specimens in PCR buffer or after phenol-chloroform extraction and subsequent ethanol precipitation. Conclusion The present study stresses the need for an internal control to confirm PCR true-negatives and demonstrates the high prevalence of chlamydiosis in Belgian turkeys and its potential zoonotic risk. The ompA nested PCR-EIA described here is a rapid, highly sensitive and specific diagnostic assay and will help to facilitate the diagnosis of Cp. psittaci infections in both poultry and man.

  9. Detection of Campylobacter in stool and determination of significance by culture, enzyme immunoassay, and PCR in developing countries.

    Science.gov (United States)

    Platts-Mills, James A; Liu, Jie; Gratz, Jean; Mduma, Esto; Amour, Caroline; Swai, Ndealilia; Taniuchi, Mami; Begum, Sharmin; Peñataro Yori, Pablo; Tilley, Drake H; Lee, Gwenyth; Shen, Zeli; Whary, Mark T; Fox, James G; McGrath, Monica; Kosek, Margaret; Haque, Rashidul; Houpt, Eric R

    2014-04-01

    Campylobacter is a common bacterial enteropathogen that can be detected in stool by culture, enzyme immunoassay (EIA), or PCR. We compared culture for C. jejuni/C. coli, EIA (ProSpecT), and duplex PCR to distinguish Campylobacter jejuni/C. coli and non-jejuni/coli Campylobacter on 432 diarrheal and matched control stool samples from infants in a multisite longitudinal study of enteric infections in Tanzania, Bangladesh, and Peru. The sensitivity and specificity of culture were 8.5% and 97.6%, respectively, compared with the results of EIA and 8.7% and 98.0%, respectively, compared with the results of PCR for C. jejuni/C. coli. Most (71.6%) EIA-positive samples were positive by PCR for C. jejuni/C. coli, but 27.6% were positive for non-jejuni/coli Campylobacter species. Sequencing of 16S rRNA from 53 of these non-jejuni/coli Campylobacter samples showed that it most closely matched the 16S rRNA of C. hyointestinalis subsp. lawsonii (56%), C. troglodytis (33%), C. upsaliensis (7.7%), and C. jejuni/C. coli (2.6%). Campylobacter-negative stool spiked with each of the above-mentioned Campylobacter species revealed reactivity with EIA. PCR detection of Campylobacter species was strongly associated with diarrhea in Peru (odds ratio [OR] = 3.66, P PCR, Campylobacter jejuni/C. coli infections represented less than half of all infections with Campylobacter species. In sum, in infants in developing country settings, the ProSpecT EIA and PCR for Campylobacter reveal extremely high rates of positivity. We propose the use of PCR because it retains high sensitivity, can ascertain burden, and can distinguish between Campylobacter infections at the species level.

  10. Effects of saliva collection using cotton swab on cortisol enzyme immunoassay.

    Science.gov (United States)

    Kozaki, Tomoaki; Hashiguchi, Nobuko; Kaji, Yumi; Yasukouchi, Akira; Tochihara, Yutaka

    2009-12-01

    Cotton swabs are among the most commonly used devices for collecting saliva, but various studies have reported that their use impacts the results of salivary cortisol assays. These studies, however, estimated this impact by comparing the average of the concentration and/or scatter plots. In the present study, we estimated the impact of cotton swabs on the results of salivary cortisol enzyme immunoassay (EIA) by Bland-Altman plot. Eight healthy males (aged 20-23 years) provided four saliva samples on different days to yield a total of 32 samples. Saliva samples were collected directly in plastic tubes using plastic straws and then pipetted onto cotton swabs (cotton saliva collection) and into clear sterile tubes (passive saliva collection). There was a lower correlation between cotton and passive saliva collection. Individually, four subjects showed a negative correlation between passive and cotton saliva collection. A Bland-Altman plot indicated that cotton swabs causes a proportional bias on the EIA assay result. Our findings indicate a considerable effect of using cotton swabs for saliva collection, and subject-specific variability in the impact. A Bland-Altman plot further suggests possible reasons for this effect.

  11. Fieldable, real-time enzyme immunoassay kits for drugs on surfaces

    Science.gov (United States)

    Chiappini, Michele W.; Wendel, Gregory J.; Duquette, Peter H.; Hamilton, Martha J.; Chudzik, Stephen J.; Chappa, Ralph A.

    1994-03-01

    Immunoassays (e.g., RIA, EIA) have been demonstrated to be useful for rapid, convenient detection and semiquantitative analysis of drugs. Thermedics Detection, Inc. manufactures a rapid, sensitive, self-contained, disposable, EIA device, developed by Bio-Metric Systems, Inc., designed to allow untrained personnel to perform in field situations. This format has been developed for drugs in biological fluids and on surfaces. The analyte in the test sample competes with an enzyme-analyte conjugate for a limited number of immobilized antibody sites. The AccuPRESS Test format can detect analytes at 10 ppb in biological fluids, water, and soil, and on surfaces, such as suitcases, vehicles, tables and hands, with positive results indicated by clearly visible color development within 5 minutes. This format is designed to have all dry components and to have an ambient shelf life of greater than one year. The format is available for cocaine and opiate derivatives, including heroin, and is readily adaptable for use with numerous other drugs, explosives, and environmental pollutants.

  12. Electrochemical enzyme-linked immunoassay in a DNA hybridization sensor

    Czech Academy of Sciences Publication Activity Database

    Paleček, Emil; Kizek, René; Havran, Luděk; Billová, Sabina; Fojta, Miroslav

    2002-01-01

    Roč. 469, č. 1 (2002), s. 73-83 ISSN 0003-2670 R&D Projects: GA ČR GV204/97/K084; GA ČR GA204/00/D049 Institutional research plan: CEZ:AV0Z5004920 Keywords : electrochemical sensor for DNA hybridization * enzyme-linked immunoassay of target DNA * magnetic beads with attached DNA probe Subject RIV: BO - Biophysics Impact factor: 2.114, year: 2002

  13. Chemiluminescence enzyme immunoassay using ProteinA-bacterial magnetite complex

    Science.gov (United States)

    Matsunaga, Tadashi; Sato, Rika; Kamiya, Shinji; Tanaka, Tsuyosi; Takeyama, Haruko

    1999-04-01

    Bacterial magnetic particles (BMPs) which have ProteinA expressed on their surface were constructed using magA which is a key gene in BMP biosynthesis in the magnetic bacterium Magnetospirillum sp. AMB-1. Homogenous chemiluminescence enzyme immunoassay using antibody bound ProteinA-BMP complexes was developed for detection of human IgG. A good correlation between the luminescence yield and the concentration of human IgG was obtained in the range of 1-10 3 ng/ml.

  14. Comparison of a Recombinant-antigen Enzyme Immunoassay with Treponema pallidum Hemagglutination Test for Serological Confirmation of Syphilis

    Directory of Open Access Journals (Sweden)

    Islay Rodríguez

    2002-04-01

    Full Text Available A recombinant-antigen enzyme immunoassay (EIA, BioSCREEN TM anti-Treponema pallidum, was compared favorably with the T. pallidum hemagglutination test, in the detection of specific antibodies in different groups of sera from patients with primary (n = 38, secondary (n = 10, early latent (n = 28 and congenital syphilis (n = 2, patients with leptospirosis ( n= 8, infectious mononucleosis (n = 7, hepatitis (n = 9, diabetes mellitus (n = 11, rheumatoid arthritis (n = 13, leprosy (n = 11, tuberculosis (n = 9, HIV/Aids ( n= 12, systemic lupus erythematosus (n = 4, rheumatic fever (n = 3, old-persons (n = 9, pregnant women (n = 29 and blood donors (n = 164. The coincidence between them was 95.1%. The sensitivity and specificity of the EIA were 93.3% and 95.5%, respectively. Fifteen serum specimens belonging to old-persons, pregnant women, blood donors, and patients with human leptospirosis, hepatitis, diabetes mellitus, tuberculosis and rheumatic fever gave false-positive results by Venereal Disease Research Laboratory and/or Rapid Plasma Reagin. The EIA can be used as alternative method for the serological confirmation of syphilis.

  15. Detection of Chlamydia trachomatis in pregnant women by the Papanicolaou technique, enzyme immunoassay and polymerase chain reaction.

    Science.gov (United States)

    Bañuelos Pánuco, C A; Deleón Rodríguez, I; Hernández Méndez, J T; Martínez Guzmán, L A; Akle Fierro, D; Miranda Murillo, J; Reyes Maldonado, E

    2000-01-01

    To compare Papanicolaou staining, enzyme immunoassay (EIA) and the polymerase chain reaction (PCR) techniques for detecting Chlamydia trachomatis in pregnant women. Endocervical specimens were taken randomly from 125 pregnant women with or without symptoms. These women attended their first medical consultation at the Regional General Ignacio Zaragoza Hospital. Samples were analyzed for detection of C trachomatis. When results differed between tests, specimens were evaluated by direct immunofluorescence staining. The prevalence of chlamydial infection was 2.4%. The characteristics of patients positive for Chlamydia were: average age, 24 years; first sexual encounter at age 21 years, one partner and six to nine months of gestation. The sensitivity, specificity, accuracy, positive predictive values and negative predictive values were 100%, 99.18%, 99.20%, 75% and 100%, respectively, for Papanicolaou staining; 100%, 92.62%, 92%, 25% and 100% for EIA; and 100%, 100%, 100% and 100% for PCR. Both Papanicolaou staining and PCR were adequate for diagnosis of C trachomatis infection. EIA was not reliable and therefore is not recommended for use as a diagnostic technique in a pregnant population with low risk and low prevalence.

  16. Development of ochratoxin a in cereal by chemiluminescence enzyme immunoassay

    Science.gov (United States)

    Li, Min; Liu, Renrong; Zhu, Lixin; Chen, Zhenzhen

    2017-11-01

    A rapid, simple and sensitive chemiluminescence enzyme immunoassay method (CLEIA) was established to detect ochratoxin A (OTA) in cereal. Optimal conditions including antibody dilution ratio and enzyme conjugate, ionic strength, pH value and organic solvent. Established indirect competition inhibition curve to determine the linear working range, detection limit and recovery rate. Results: The 50% inhibitory concentration and the detection limit of the CLEIA were78.8pg/mL and 14.86 pg/mL, respectively, with a linear range of 0.015-0.4ng/mL. At 1∼4μpg/kg fortified levels in wheat, mean recoveries ranged from 67.47% to100.35%.

  17. Diagnosis of acute radiation disease by Enzyme Immune-Assay (EIA)

    International Nuclear Information System (INIS)

    Popov, D.; Maliev, V.; Jones, J.; Gonta, S.; Prasad, K.; Rachal, C.

    2006-01-01

    Diagnosis of the acute radiation disease by the method of immune enzyme assay is a simple and efficient tool of evaluating and biological dosimetry and forecasting of development of the acute radiation defeats as at group of population so at individuals locating in the zone polluted by the radiation. We use as biological markers the group of essential radiotoxins - high molecular mass glycoprotein ( molecular mass - 200 - 250 kDa ) - radiation antigens (S.D.R. - specific radiation determinant ) accumulated in the lymphoid system, with epitopes specific to each form of radiation syndrome, after animals have been irradiated in doses inducing the development of the cerebral (1), toxic ( 2), gastrointestinal ( 3 ) and typical ( 4 ) forms of acute radiation sickness. These two phenomena allowed us to develop a technologies for diagnosis, prophylaxis and therapy of radiation disease - enzyme immune assay ( EIA ), anti radiation vaccine, anti radiation serum, method of immune - lymph - plasma-sorption. The important first step in effectiveness of therapy is an accurate assessment of severity of disease in early period after irradiation. The ideal markers for early and accurate assessment is high weight glycoprotein with specifics radiation induced features (S.D.R.) mentioned above. This biology active substance isolated from lymph can induct the symptoms of radiation syndrome without previously radiation when it is administrated intra-muscularly or intravenously to healthy animals. Enzyme immune assay (EIA) allowed researchers to indicate the significant levels of different forms of S.D.R. in peripheral blood of animals in first 24 hours after radiation. Indication of high level of S.D.R. -1 allowed to forecast a fast development of cerebral form of acute radiation disease. Determination of high levels of S.D.R.-2, S.D.R.-3 and S.D.R.-4 in peripheral blood allowed to recognize early periods of toxic, gastrointestinal and typical forms of acute radiation sickness

  18. Diagnosis of acute radiation disease by enzyme immune-assay (EIA)

    International Nuclear Information System (INIS)

    Popov, D.; Maliev, V.

    2005-01-01

    Full text: Multifactor fundamental research under experimental and industrial conditions on farm and laboratory animals and also on humans blood serum of Chernobyl NPP clean-up workers conducted within 1982 - 2002 have enabled us to establish the existence of two previously unknown phenomena: the phenomenon of reversible redistribution of cytobiochemical parameters in the blood-interstices-lymph-blood system of irradiated animals, which supports compensatory maintenance of homeostasis. The phenomenon of specific immunochemical reactions to the radiological effect, involving the formation of high molecular mass glycoprotein (molecular mass - 200 - 250 kDa) - radiation antigens (SDR - specific radiation determinant) in the lymphoid system with epitopes specific to each form of radiation syndrome, after animals have been irradiated in doses inducing the development of the cerebral (1) , toxic (2) gastrointestinal (3) and typical (4) forms of acute radiation sickness. These two phenomena allowed us to develop a technologies for diagnosis, prophylaxis and therapy of radiation disease - enzyme immune assay (EIA), antiradiation vaccine, antiradiation serum, method of immune-lymph-plasmosorption. The important first step in effectiveness of therapy is an accurate assessment of severity of disease in early period after irradiation. The ideal markers for early and accurate assessment is high weight glycoprotein with specifics radiation induced features (SDR) mentioned above. This biology active substance isolated from lymph can induct the symptoms of radiation syndrome without previously radiation when it is administrated intramuscularly or intravenously to healthy animals. Enzyme immune assay (EIA) allowed researchers to indicate the significant levels of different forms of SDR in peripheral blood of animals in first 24 hours after radiation. Indication of high level of SDR -1 allowed to forecast a fast development of cerebral form of acute radiation disease. Determination

  19. Comparative determination of phenytoin by spectrophotometry, gas chromatography, liquid chromatography, enzyme immunoassay, and radioimmunoassay

    International Nuclear Information System (INIS)

    Castro, A.; Ibanez, J.; DiCesare, J.L.; Adams, R.F.; Malkus, H.

    1978-01-01

    Sera from patients being treated with phenytoin were analyzed for the drug by spectrophotometry, gas chromatography, radioimmunoasay, enzyme immunoassay, and liquid chromatography. The assay values obtained were intercompared statistically. Enzyme immunoassay and liquid chromatography appear to be attractive alternatives to the more traditional methods of spectrophotometry and gas chromatography. Our radioimmunoassay data correlated poorly with results by the four other methods

  20. Large-scale evaluation of the immuno-mycologics lateral flow and enzyme-linked immunoassays for detection of cryptococcal antigen in serum and cerebrospinal fluid.

    Science.gov (United States)

    Hansen, Jessica; Slechta, E Susan; Gates-Hollingsworth, Marcellene A; Neary, Brandon; Barker, Adam P; Bauman, Sean; Kozel, Thomas R; Hanson, Kimberly E

    2013-01-01

    Cryptococcosis is a systemic infection caused by the pathogenic yeasts Cryptococcus neoformans and C. gattii. Detection of cryptococcal capsular antigen (CrAg) in serum and cerebrospinal fluid (CSF) plays an important diagnostic role. We prospectively compared the new Immuno-Mycologics Inc. (IMMY) lateral flow assay (LFA) and enzyme immunoassay (EIA) to our current CrAg test (Premier EIA; Meridian Bioscience Inc.). Discordant samples were retested with the latex-Cryptococcus antigen test (IMMY) and using serotype-specific monoclonal antibodies (MAbs). A total of 589 serum and 411 CSF specimens were tested in parallel. Qualitative agreement across assays was 97.7%. In all, 56 (41 serum and 15 CSF) samples were positive and 921 (527 serum and 394 CSF) samples were negative by all three assays. The 23 discrepant specimens were all Meridian EIA negative. Of 23 discordant specimens, 20 (87.0%) were positive by both the IMMY LFA and EIA, 2 were LFA positive only, and 1 was EIA positive only. Eleven discrepant specimens had adequate volume for latex agglutination (LA) testing; 8 were LA positive, and 3 were LA negative. LA-negative samples (2 CSF samples and 1 serum) had low IMMY LFA/EIA titers (≤1:10). Serotype-specific MAb analysis of the LA-positive samples suggested that these specimens contained CrAg epitopes similar to those of serotype C strains. In conclusion, the IMMY assays showed excellent overall concordance with the Meridian EIA. Assay performance differences were related to issues of analytic sensitivity and possible serotype bias. Incomplete access to patient-level data combined with low specimen volumes limited our ability to fully resolve discrepant results.

  1. Development of a novel ultrasensitive enzyme immunoassay for human glutamic acid decarboxylase 65 antibody.

    Science.gov (United States)

    Numata, Satoshi; Katakami, Hideki; Inoue, Shinobu; Sawada, Hirotake; Hashida, Seiichi

    2016-07-01

    We developed a novel, ultrasensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) for determination of glutamic acid decarboxylase autoantibody concentrations in serum samples from patients with type 2 diabetes. We developed an immune complex transfer enzyme immunoassay for glutamic acid decarboxylase autoantibody and measured glutamic acid decarboxylase autoantibody from 22 patients with type 1 diabetes, 29 patients with type 2 diabetes, and 32 healthy controls. A conventional ELISA kit identified 10 patients with type 1 diabetes and one patient with type 2 diabetes as glutamic acid decarboxylase autoantibody positive, whereas 15 patients with type 1 diabetes and six patients with type 2 diabetes were identified as glutamic acid decarboxylase autoantibody positive using immune complex transfer enzyme immunoassay. Immune complex transfer enzyme immunoassay is a highly sensitive and specific assay for glutamic acid decarboxylase autoantibody and might be clinically useful for diabetic onset prediction and early diagnosis. © The Author(s) 2016.

  2. Rapid determination of recent cocaine use with magnetic particles-based enzyme immunoassays in serum, saliva, and urine fluids.

    Science.gov (United States)

    Vidal, Juan C; Bertolín, Juan R; Bonel, Laura; Asturias, Laura; Arcos-Martínez, M Julia; Castillo, Juan R

    2016-06-05

    Cocaine is one of the most worldwide used illicit drugs. We report a magnetic particles-based enzyme-linked immunoassay (mpEIA) method for the rapid and sensitive determination of cocaine (COC) in saliva, urine and serum samples. Under optimized conditions, the limits of detections were 0.09ngmL(-1) (urine), 0.15ngmL(-1) (saliva), and 0.06ngmL(-1) COC (human serum). Sensitivities were in the range EC50=0.6-2.5ngmL(-1) COC. The cross-reactivity with the principal metabolite benzoylecgonine (BZE) was only 1.6%. Recovering percentages of doped samples (0, 10, 50, and 100ngmL(-1) of COC) ranged from about 86-111%. Some advantages of the developed mpEIA over conventional ELISA kits are faster incubations, improved reproducibility, and consumption of lower amounts of antibody and enzyme conjugates due to the use of magnetic beads. The reported method was validated following the guidelines on bioanalytical methods of the European Medicines Agency (2011). Unmetabolized COC detection has a great interest in pharmacological, pharmacokinetics, and toxicokinetics studies, and can be used to detect a very recent COC use (1-6h). Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Quality assurance in immunoassay performance--comparison of different enzyme immunoassays for the determination of caffeine in consumer products.

    Science.gov (United States)

    Grandke, Julia; Oberleitner, Lidia; Resch-Genger, Ute; Garbe, Leif-Alexander; Schneider, Rudolf J

    2013-02-01

    Enzyme immunoassays with optical detection are amongst the most widely used bioanalytical tools. We defined seven parameters for the quality assessment of immunoassays that were addressed in a systematic study of direct and indirect immunoassays, using the enzymes horseradish peroxidase (HRP) and alkaline phosphatase (AP), the chromogenic substrates 3,3',5,5'-tetramethylbenzidine (TMB) and para-nitrophenyl phosphate, and the fluorescent substrates 3-(4-hydroxyphenyl)propionic acid and 4-methylumbelliferyl phosphate. The same monoclonal antibody against caffeine was used throughout the study. The four quality parameters regarding the standard curve were the test midpoint (sensitivity), the measurement range, the relative dynamic range of the signal, and the goodness of fit of the adjusted four-parameter logistic function. All HRP immunoassays showed a higher sensitivity compared to the AP assays. On the basis of all four criteria, it was established that the direct assay format is superior to the indirect format, the immunoassay using HRP TMB fulfilling all requirements best. In a second step, caffeine concentrations in 24 beverage and cosmetics samples were determined and three more quality parameters were assessed with this application. The direct HRP TMB assay showed one of the best intra- and inter-plate precisions and the best accuracy, defined by the correlation of results with those from the chosen reference method liquid chromatography tandem mass spectrometry (LC-MS/MS). Considering all criteria, HRP TMB seems to be the enzyme substrate system of choice preferably used in the direct assay format.

  4. Comparison of a frozen human foreskin fibroblast cell assay to an enzyme immunoassay and toxigenic culture for the detection of toxigenic Clostridium difficile.

    Science.gov (United States)

    Strachan, Alastair J; Evans, Natalie E; Williams, O Martin; Spencer, Robert C; Greenwood, Rosemary; Probert, Chris J

    2013-01-01

    This study set out to validate the Hs27 ReadyCell assay (RCCNA) as an alternative CCNA method compared against a commonly used commercial enzyme immunoassay (EIA) method and toxigenic culture (TC) reference standard. A total of 860 samples were identified from those submitted to the Health Protection Agency microbiology laboratories over a 30-week period. RCCNA performed much better than EIA when using TC as a gold standard, with sensitivities of 90.8% versus 78.6% and positive predictive value of 87.3% to 81.9%, respectively. The Hs27 Human Foreskin Fibroblast ReadyCells are an easy-to-use and a sensitive CCNA method for the detection of toxigenic Clostridium difficile directly from stool. A turnaround time of up to 48 h for a negative result and possible need for repeat testing make it an unsuitable method to be used in most clinical laboratory setting. Copyright © 2013 Elsevier Inc. All rights reserved.

  5. Comparison of a frozen human foreskin fibroblast cell assay to an enzyme immunoassay and toxigenic culture for the detection of toxigenic Clostridium difficile☆☆☆★

    Science.gov (United States)

    Strachan, Alastair J.; Evans, Natalie E.; Williams, O. Martin; Spencer, Robert C.; Greenwood, Rosemary; Probert, Chris J.

    2013-01-01

    This study set out to validate the Hs27 ReadyCell assay (RCCNA) as an alternative CCNA method compared against a commonly used commercial enzyme immunoassay (EIA) method and toxigenic culture (TC) reference standard. A total of 860 samples were identified from those submitted to the Health Protection Agency microbiology laboratories over a 30-week period. RCCNA performed much better than EIA when using TC as a gold standard, with sensitivities of 90.8% versus 78.6% and positive predictive value of 87.3% to 81.9%, respectively. The Hs27 Human Foreskin Fibroblast ReadyCells are an easy-to-use and a sensitive CCNA method for the detection of toxigenic Clostridium difficile directly from stool. A turnaround time of up to 48 h for a negative result and possible need for repeat testing make it an unsuitable method to be used in most clinical laboratory setting. PMID:23107315

  6. Immunoassays

    Science.gov (United States)

    Hsieh, Y.-H. Peggy

    Immunochemistry is a relatively new science that has developed rapidly in the last few decades. One of the most useful analytical developments associated with this new science is immunoassay. Originally immunoassays were developed in medical settings to facilitate the study of immunology, particularly the antibody-antigen interaction. Immunoassays now are finding widespread applications outside the clinical field because they are appropriate for a wide range of analytes ranging from proteins to small organic molecules. In the food analysis area, immunoassays are widely used for chemical residue analysis, identification of bacteria and viruses, and detection of proteins in food and agricultural products. Protein detection is important for determination of allergens and meat species content, seafood species identification, and detection of genetically modified plant tissues. While immunoassays of all formats are too numerous to cover completely in this chapter, there are several procedures that have become standard for food analysis because of their specificity, sensitivity, and simplicity.

  7. Two competitive enzyme immunoassays for the detection of IgG class antibodies to hepatitis a antigen

    Directory of Open Access Journals (Sweden)

    Claudia Lamarca Vitral

    1991-06-01

    Full Text Available Two competitive enzyme immunoassays (EIA techniques were developed: in the first (COMP-1, test sera were added together with HAV antigen on anti-HAV IgG-coated wells followed by an anti-HA VHRP conjugate; in the second (COMP-2, test sera and anti-HA VHRP conjugate competed for HAV epitopes previously adsorbed to anti-HA V IgG-coated wells. Both procedures used tetramethylbenzidine (TMB as a substrate. Both competitive tests were shown to be reproducible and suitable for routine diagnosis and research purposes.Foram desenvolvidos dois ensaios imunoenzimáticos (EIA competitivos: no primeiro (COMP-1 colocou-se numa placa sensibilizada com anti-HAVIgG as amostras teste juntamente como antígeno HA Vea seguir o conjugado anti-HA VHRP; no segundo (COMP-2, as amostras teste e o conjugado anti-HAV HRP competem pelos epitopos do antígeno HAV previamente absorvido na placa sensibilizada do anti-HAV IgG. O substrato utilizado foi tetrametilbenzidina (TMB. Ambas as técnicas mostraram ser produtíveis e aplicáveis para fins de diagnóstico e pesquisa.

  8. Validation of an enzyme-immunoassay for the non-invasive monitoring of faecal testosterone metabolites in male cheetahs (Acinonyx jubatus).

    Science.gov (United States)

    Pribbenow, Susanne; Wachter, Bettina; Ludwig, Carsten; Weigold, Annika; Dehnhard, Martin

    2016-03-01

    In mammals, the sex hormone testosterone is the major endocrine variable to objectify testicular activity and thus reproductive function in males. Testosterone is involved in the development and function of male reproductive physiology and sex-related behaviour. The development of a reliable androgen enzyme-immunoassay (EIA) to monitor faecal testosterone metabolites (fTM) is a powerful tool to non-invasively assess the gonadal status of males. We validated an epiandrosterone EIA for male cheetahs by performing a testosterone radiometabolism study followed by high-performance liquid chromatography (HPLC) analyses and excluding possible cross-reactivities with androgenic metabolites not derived from testosterone metabolism. The physiological and biological relevance of the epiandrosterone EIA was validated by demonstrating (1) a significant increase in fTM concentrations within one day in response to a testosterone injection, (2) a significant increase in fTM concentrations within one day in response to a gonadotropin-releasing hormone (GnRH) injection, which failed following a placebo injection, and (3) significant differences in fTM concentrations between adult male and adult female cheetahs and between adult and juvenile male cheetahs of a free-ranging population. Finally, we demonstrated stability of fTM concentrations measured in faecal samples exposed to ambient temperatures up to 72h. Our results clearly demonstrate that the epiandrosterone EIA is a reliable non-invasive method to monitor testicular activity in male cheetahs. Copyright © 2016 Elsevier Inc. All rights reserved.

  9. A multicenter evaluation of the Biotest legionella urinary antigen EIA

    DEFF Research Database (Denmark)

    Harrison, Timothy; Uldum, Søren; Alexiou-Daniel, Stella

    1998-01-01

    OBJECTIVES: To undertake a multicenter study to evaluate the Biotest legionella urinary antigen enzyme immunoassay (EIA) performance against those EIAs already in use in 14 European laboratories. METHODS: Each laboratory examined urine specimens from appropriate patients using both their current...... assay and the Biotest EIA. Each examined: a standard panel of 12 coded urine samples (distributed by Biotest); a panel of 10 coded urine samples provided as part of a European external quality assurance (EQA) scheme; urine samples from patients with proven legionnaires' disease (LD); urine samples from...... patients with pneumonia of microbiologically proven cause other than LD; and urine samples submitted for routine examination. Thus, the performance of the Biotest assay (in comparison with current EIAs), its specificity and utility, and the inter-laboratory agreement were assessed. RESULTS: Inter...

  10. Comparison of enzyme immunoassays for the diagnosis of bovine brucellosis

    International Nuclear Information System (INIS)

    Nielsen, K.; Kelly, L.; Gall, D.; Balsevicius, S.; Bosse, J.; Kelly, W.; Nicoletti, P.

    1998-01-01

    The indirect enzyme immunoassay for measurement of bovine antibody to Brucella abortus was tested on 15,716 Canadian sera to assess the specificity. These sera were also tested by the buffered plate antigen test. Two ELISA formats were used for assessment of data: the targeting procedure using a positive control serum allowed to develop to an optical density of 1.0 and the use of a positive control serum to determine relative positivity at a set time. Two different cut-off values were also assessed for each assay. A total of 763 sera gave reactions above established cut-off values in the ELISA while 216 were positive in the buffered plate antigen test (BPAT). A modification of the indirect ELISA employed divalent cation chelating agents (EDTA/EGTA) incorporated into the serum incubation stage to eliminate some non-specific reactions. This method was applied only to the 763 indirect ELISA reactor sera and it eliminated all but 93 or 37, depending on the cut-off selected, of the reactions. Sensitivity was assessed by testing 424 sera from Brucella abortus culture positive cattle. The indirect ELISA classified all 424 sera as positive by either method of data handling and with or without addition of EDTA/EGTA for a specificity estimate of 100%. In the BPAT, 412 sera gave a positive agglutination reaction. Ten percent of the 15,716 sera were randomly selected and tested by two different competitive ELISAs and by the complement fixation test (CFT). One competitive ELISA used Brucella abortus O-polysaccharide as the antigen and an enzyme conjugated monoclonal antibody to the O-polysaccharide for competition and detection. Of the sera tested, 34 gave false positive reactions. On a retest, the false positive reactions were reduced to 2. The second competitive ELISA used lipopolysaccharide as the antigen, a different monoclonal antibody but also specific for the O-polysaccharide for competition and commercially available goat anti-mouse IgG enzyme conjugate for detection

  11. Evaluation of six immunoassays for detection of dengue virus-specific immunoglobulin M and G antibodies

    NARCIS (Netherlands)

    J. Groen (Jan); P. Koraka (Penelope); J. Velzing (Jans); C. Copra (Cederick); A.D.M.E. Osterhaus (Albert)

    2000-01-01

    textabstractThe performance of six commercially available immunoassay systems for the detection of dengue virus-specific immunoglobulin M (IgM) and IgG antibodies in serum was evaluated. These included two IgM and IgG enzyme immunoassays (EIA) from MRL Laboratories and PanBio, a rapid

  12. Diagnosing toxigenic Clostridium difficile: new confidence bounds show culturing increases sensitivity of the toxin A/B enzyme immunoassay and refute gold standards.

    Science.gov (United States)

    Mattner, Frauke; Winterfeld, Ingo; Mattner, Lutz

    2012-08-01

    To scrutinize published sensitivity estimates obtained using questionable gold standards by comparing sensitivities of culturing Clostridium difficile in commercially available media followed by enzyme immunoassay (EIA) toxin A or B detection (culture test) with applying the EIA to stool samples alone (direct test). In 2008, consecutive stool samples were cultured on C. difficile selective culture media: (1) medium I: Clostridium difficile-selective agar (CDSA; Becton Dickinson); (2) medium II: CLO agar (BioMérieux); (3) medium III: C. difficile agar according to Brazier (Oxoid). In addition, a direct test was performed (Ridascreen, r-Biopharm), which was also used to confirm toxin A or B production in the cultured C. difficile. New confidence bounds for sensitivities were applied, without assuming any perfect reference test or any conditional independence of the tests compared. Of 256 liquid stool samples, 18.4% were diagnosed as positive by at least 1 of the 4 tests; 12.8% were positive with culture medium I, 16.4% with medium II, and 13.6% with medium III, and 10.1% were positive by the direct test. Assuming culture tests to be at least as specific as the direct test yields an upper bound of 61% (upper 95% confidence bound (CB) 81%) for the sensitivity of the direct test. Assuming a prevalence of 15% yields sensitivity gains of the culture tests of at least 18% (lower 95% CB--4%) for medium I, 40% (lower 95% CB 21%) for medium II, and 23% (lower 95% CB 2%) for medium III. Published high sensitivities of direct toxin A/B EIAs, up to 96%, and the correctness of the cytotoxicity test assumed for their estimation are doubtful. With culture medium II, sensitivity gains of at least about 20% are obtainable. Direct toxin A/B EIAs alone are insufficiently sensitive for the clinical diagnosis of C. difficile infections.

  13. Challenges, pitfalls and surprises: development and validation of a monoclonal antibody for enzyme immunoassay of the steroid 1α-hydroxycorticosterone in elasmobranch species.

    Science.gov (United States)

    Wheaton, Catharine J; Mylniczenko, Natalie D; Rimoldi, John M; Gadepalli, Rama S V S; Hart, R; O'Hara, Bobbi R; Evans, Andrew N

    2018-01-31

    Sharks and rays are popular species used in wildlife ecotourism and aquariums to educate the public on the behavior, ecology and conservation challenges of elasmobranchs. To understand long-term physiological health and welfare under varying social and husbandry conditions, we developed and validated an enzyme immunoassay (EIA) to measure stress/ionoregulatory hormones in managed and semi-free range southern rays (Hypanus americanus). Banked serum and interrenal samples from 27 female rays managed at Disney's The Seas with Nemo and Friends® and Castaway Cay were used to evaluate measurement of 1α-hydroxycorticosterone (1αOHB) relative to corticosterone (B). Although commercial EIAs are available for B, those tested exhibit only low relative cross-reactivity to 1αOHB (3-5%). To improve measurement of 1αOHB, we developed a monoclonal antibody using a synthesized 1αOHB-derivative for evaluation using high-performance liquid chromatography (HPLC) and EIA. Relative displacements of cross-reactant compounds showed that the antibody had good sensitivity for the target antigen 1αOHB, and low sensitivity to related steroids (desoxycorticosterone and B), but greater sensitivity to 11-dehydrocorticosterone. Tests of competitive vs. noncompetitive EIA formats, reagent titration, and incubation times of the antibody and conjugate were used to optimize sensitivity, repeatability and precision of measured 1αOHB in standards and samples (4 ng/ml, 90% binding). Tests of sample pre-treatment (pH adjustment) and extraction with varying solvent polarity were used to optimize measurement of 1αOHB in <1 ml (serum) or 1 g (interrenal) samples. HPLC analysis revealed the 1αOHB EIA to be superior for measurement of 1αOHB compared to use of a B EIA with or without HPLC fractioning. Results may prove useful for extrapolation to guide best practices for 1αOHB measurement in other elasmobranch species. Improved measurement of stress/ionoregulatory hormones in sharks and rays

  14. False-negative syphilis treponemal enzyme immunoassay results in an HIV-infected case-patient.

    Science.gov (United States)

    Katz, Alan R; Komeya, Alan Y; Tomas, Juval E

    2017-06-01

    We present a case report of a false-negative syphilis treponemal enzyme immunoassay test result in an HIV-infected male. While treponemal tests are widely considered to be more sensitive and specific than non-treponemal tests, our findings point to potential challenges using the reverse sequence syphilis screening algorithm.

  15. Anti-bovine antibody in human sera as a cause of nonspecificity in enzyme immunoassay.

    OpenAIRE

    Dise, T; Brunell, P A

    1987-01-01

    Much nonspecific activity in enzyme immunoassay is due to reactions between anti-bovine antibodies which are commonly present in human serum samples and residual calf serum in viral antigens. Calf serum is used in cultured cells which are used to produce viral antigens. Addition of bovine proteins to serum samples to be tested for viral antibodies reduces this activity.

  16. Competitive enzyme immunoassay for human chorionic somatomammotropin using the avidin-biotin system

    International Nuclear Information System (INIS)

    Rappuoli, R.; Leoncini, P.; Tarli, P.; Neri, P.

    1981-01-01

    Human chorionic somatomammotropin (HCS) is determined by an enzyme immunoassay where HCS competes with biotin-labeled HCS for insolubilized anti-HCS antibodies. Enzyme-labeled avidin is then used to reveal the amount of bound HCS. The system proves to be sensitive (1 ng/ml of HCS can be detected) and results agree with radioimmunoassay determinations (correlation coefficient = 0.979). Kinetics of the avidin-biotin reaction and coating of polystyrene wells are also investigated

  17. The effect of storage conditions on salivary cortisol concentrations using an enzyme immunoassay

    DEFF Research Database (Denmark)

    Nalla, Anjana A; Thomsen, Gerda; Knudsen, Karen Birgitte Moos

    2015-01-01

    Saliva samples are easy to collect and are applicable for home-sampling, e.g. when studying HPA-axis dynamics to characterize diurnal cortisol profiles and the cortisol awakening response. However, the storing and transport conditions might be critical in the home-sampling approach. Here, we tested...... the stability of saliva cortisol in samples stored at different temperatures and after repeated thawing-freezing cycles when measured with an Enzyme Immuno Assay (EIA). Thirteen healthy volunteers, six women and seven men, mean age 31 (range 26-49) years collected saliva either in the morning hours (08...... it was stored at - 80°C. The last tube was stored directly at - 80°C and served as the 'gold standard'. The saliva samples were assayed using Salivary Cortisol Diagnostic EIA. Differences in cortisol measurements between each of the five conditions and the 'gold standard' (- 80°C) were evaluated by one-sample t...

  18. Enhanced lateral flow immunoassay using gold nanoparticles loaded with enzymes.

    Science.gov (United States)

    Parolo, Claudio; de la Escosura-Muñiz, Alfredo; Merkoçi, Arben

    2013-02-15

    The use of gold nanoparticles (AuNPs) as labeling carriers in combination with the enzymatic activity of the horseradish peroxidase (HRP) in order to achieve an improved optical lateral flow immunoassay (LFIA) performance is presented here. Briefly in a LFIA with an immune-sandwich format AuNPs are functionalized with a detection antibody already modified with HRP, obtaining an 'enhanced' label. Two different detection strategies have been tested: the first one following just the red color of the AuNPs and the second one using a substrate for the HRP (3 different substrates are evaluated), which produces a darker color that enhances the intensity of the previous red color of the unmodified AuNPs. In such very simple way it is gaining sensitivity (up to 1 order of magnitude) without losing the simplicity of the LFIA format, opening the way to other LFIA applications including their on-demand performance tuning according to the analytical scenario. Copyright © 2012 Elsevier B.V. All rights reserved.

  19. An enzyme immunoassay for detection of Japanese encephalitis ...

    Indian Academy of Sciences (India)

    The chemotactic factor could be detected in JEV inoculated mouse sera and JEV infected culture fluids. Significant finding of the test was the detection of hMDF in sera of human cases of JE. 1. Introduction. Enzyme linked immunosorbent assay (ELISA) is a highly sensitive technique which is specific, less time consuming.

  20. Detection of phase I IgG antibodies to Coxiella burnetii with EIA as a screening test for blood donations

    NARCIS (Netherlands)

    van der Hoek, W.; Wielders, C. C. H.; Schimmer, B.; Wegdam-Blans, M. C. A.; Meekelenkamp, J.; Zaaijer, H. L.; Schneeberger, P. M.

    2012-01-01

    The presence of a high phase I IgG antibody titre may indicate chronic infection and a risk for the transmission of Coxiella burnetii through blood transfusion. The outbreak of Q fever in the Netherlands allowed for the comparison of an enzyme immunoassay (EIA) with the reference immunofluorescence

  1. Immune status of health care workers to measles virus: evaluation of protective titers in four measles IgG EIAs

    NARCIS (Netherlands)

    Dorigo-Zetsma, J.W.; Hall, M.A.; Vreeswijk, J.; Vries, J.J. de; Vossen, A.C.; Hulscher, H.I. Ten; Kerkhof, J.; Smits, G.P.; Ruijs, W.L.M.; Koopmans, M.P.; Binnendijk, R.S. van

    2015-01-01

    BACKGROUND: Following the recognition of a measles case in a hospital in The Netherlands, health care workers (HCW) from the premises were screened by a commercial enzyme immunoassay (EIA) for measles IgG to identify persons at risk for measles. At least 10% of the HCW were tested measles

  2. Use of inorganic salts to minimize serum interference in a sandwich enzyme immunoassay for human growth hormone using Fab'-horseradish peroxidase conjugate.

    Science.gov (United States)

    Hashida, S; Nakagawa, K; Imagawa, M; Inoue, S; Yoshitake, S; Ishikawa, E; Endo, Y; Ohtaki, S; Ichioka, Y; Nakajima, K

    1983-12-30

    In a sandwich enzyme immunoassay (EIA) for human growth hormone (hGH) with anti-hGH Fab'-peroxidase conjugate, the effects of inorganic salts on serum interference were examined, and serum interference was eliminated by incubation of serum samples with anti-hGH IgG-coated polystyrene balls in the presence of 0.4 mol/l NaCl, avoiding the need for hGH-free serum. The sensitivity for hGH was 60 fg/tube or 3 ng/l of serum. No cross-reaction was observed with prolactin, chorionic gonadotropin or luteinizing, thyroid-stimulating and follicle-stimulating hormones. The coefficients of within-assay and between-assay variations were 2.8-6.5% and 4.8-8.7%, respectively. The regression equation and correlation coefficient to radioimmunoassay (RIA) were y(EIA) = 0.89x(RIA) + 0.11 and 0.98 (n = 100), respectively. hGH levels in normal male and female adult serum taken between 9:00 and 10:00 a.m. after overnight fasting and 1 h rest were 312 ng/l (range 53-940 ng/l; n = 10) and 662 ng/l (112-2195 ng/l; n = 13), respectively.

  3. Development and application of radioimmunoassay and enzyme immunoassays in microbiological and immunological diagnosis. 4

    International Nuclear Information System (INIS)

    Struy, H.; Horeschi, G.; Wolffgang, H.; Morenz, J.

    1982-01-01

    Indirect solid phase radioimmunoassays and enzyme immunoassays were developed for the quantitative determination of low concentrations of IgG, IgA, IgM, and IgE. PVC blister and microtiterplates of polystyrene with adsorptively bound purified immunoglobulins were used as carrier. The determination of the immunoglobulins was accomplished with a uniform system of indicators, which consists of class-specific antihuman immunoglobulin sera from rabbits and of labelled ( 125 I and horseradish peroxydase, resp.) antirabbitglobulin from sheep. The developped competitive radioimmuno- and enzyme immunoassays allows the determination of concentrations of classes of immunoglobulins down to nearly 1 ng/ml, and the indirect sandwich solid phase radioimmunoassay determinations of IgE down to about 1 IU/ml. The values of IgG determined in liquors and those of IgE found in sera correspond very well to values which were measured with commercial kits. (author)

  4. Cytomegalovirus Antibody in Cerebrospinal Fluid of Schizophrenic Patients Detected by Enzyme Immunoassay

    Science.gov (United States)

    Fuller Torrey, E.; Yolken, Robert H.; Winfrey, C. Jack

    1982-05-01

    By means of enzyme immunoassay techniques to detect the presence of antibody to cytomegalovirus, the cerebrospinal fluid of 178 patients with schizophrenia, 17 patients with bipolar disorders, and 11 other psychiatric patients was compared with that of 79 neurological patients and 41 normal control subjects. The cerebrospinal fluid of 20 of the schizophrenic patients and 3 of the patients with bipolar disorders showed significant increases in immunoglobulin M antibody to cytomegalovirus; no difference was found in patients on or off psychotropic medications.

  5. Utility of salivary enzyme immunoassays for measuring estradiol and testosterone in adolescents: a pilot study.

    Science.gov (United States)

    Amatoury, Mazen; Lee, Jennifer W; Maguire, Ann M; Ambler, Geoffrey R; Steinbeck, Katharine S

    2016-04-09

    We investigated the utility of enzyme immunoassay kits for measuring low levels of salivary estradiol and testosterone in adolescents and objectively assessed prevalence of blood contamination. Endocrine patients provided plasma and saliva for estradiol (females) or testosterone (males) assay. Saliva samples were also tested with a blood contamination kit. Picomolar levels of salivary estradiol in females failed to show any significant correlation with plasma values (r=0.20, p=0.37). The nanomolar levels of salivary testosterone in males showed a strong correlation (r=0.78, p<0.001). A significant number of saliva samples had blood contamination. After exclusion, correlations remained non-significant for estradiol, but strengthened for testosterone (r=0.88, p<0.001). The salivary estradiol enzyme immunoassay is not clinically informative at low levels. Users should interpret clinical saliva with caution due to potential blood contamination. Our data supports the utility of the salivary testosterone enzyme immunoassay for monitoring adolescent boys on hormone developmental therapy.

  6. Enzyme immunoassay of oestrogen receptors in needle biopsies from human liver

    DEFF Research Database (Denmark)

    Becker, U; Andersen, J; Poulsen, H S

    1991-01-01

    For quantitative assessments of sex hormone receptors in liver tissue, ligand binding assays are inconvenient, as they require large biopsies (0.5-1.0 g). The present study shows that it is possible to measure oestrogen receptors (ER) quantitatively in needle biopsy specimens as small as 10 mg...... by modifications of a commercial enzyme immunoassay employing monoclonal antibodies. Sucrose gradient centrifugation and the dextran charcoal method served as reference methods. A consecutive series of needle biopsies from patients suspected of liver disease were investigated. The biopsies (n = 37) had a median...

  7. Clostridium difficile testing algorithms using glutamate dehydrogenase antigen and C. difficile toxin enzyme immunoassays with C. difficile nucleic acid amplification testing increase diagnostic yield in a tertiary pediatric population.

    Science.gov (United States)

    Ota, Kaede V; McGowan, Karin L

    2012-04-01

    We evaluated the performance of the rapid C. diff Quik Chek Complete's glutamate dehydrogenase antigen (GDH) and toxin A/B (CDT) tests in two algorithmic approaches for a tertiary pediatric population: algorithm 1 entailed initial testing with GDH/CDT followed by loop-mediated isothermal amplification (LAMP), and algorithm 2 entailed GDH/CDT followed by cytotoxicity neutralization assay (CCNA) for adjudication of discrepant GDH-positive/CDT-negative results. A true positive (TP) was defined as positivity by CCNA or positivity by LAMP plus another test (GDH, CDT, or the Premier C. difficile toxin A and B enzyme immunoassay [P-EIA]). A total of 141 specimens from 141 patients yielded 27 TPs and 19% prevalence. Sensitivity, specificity, positive predictive value, and negative predictive value were 56%, 100%, 100%, and 90% for P-EIA and 81%, 100%, 100%, and 96% for both algorithm 1 and algorithm 2. In summary, GDH-based algorithms detected C. difficile infections with superior sensitivity compared to P-EIA. The algorithms allowed immediate reporting of half of all TPs, but LAMP or CCNA was required to confirm the presence or absence of toxigenic C. difficile in GDH-positive/CDT-negative specimens.

  8. The management of isolated positive syphilis enzyme immunoassay results in HIV-negative patients attending a sexual health clinic.

    Science.gov (United States)

    Thorley, Nicola; Adebayo, Michael; Smit, Erasmus; Radcliffe, Keith

    2016-08-01

    An unconfirmed positive treponemal enzyme immunoassay (enzyme immunoassay positive, Treponema pallidum particle agglutination negative and rapid plasma reagin negative) presents a clinical challenge to distinguish early syphilis infection from false-positive results. These cases are referred for syphilis line assay (INNO-LIA) and recalled for repeat syphilis serology. We performed a retrospective audit to establish the proportion of HIV-negative cases with unconfirmed positive enzyme immunoassay results, the proportion of these cases that received an INNO-LIA test and repeat syphilis serology testing and reviewed the clinical outcomes; 0.35% (80/22687) cases had an unconfirmed positive treponemal enzyme immunoassay result. Repeat syphilis serology was performed in 80% (64/80) cases, but no additional cases of syphilis were identified. Eighty-eight per cent (70/80) received an INNO-LIA test; 14% (5/37) unconfirmed enzyme immunoassay-positive cases with no prior history of syphilis were confirmed on INNO-LIA assay, supporting a diagnosis of latent syphilis. As a confirmatory treponemal test, the INNO-LIA assay may be more useful than repeat syphilis serological testing. © The Author(s) 2016.

  9. Determination of serum digosin. A comparison between RIA and EIA

    International Nuclear Information System (INIS)

    Spitz, J.; Braun, J.S.; Schmidt, M.; Krankenhausstiftung Bamberg

    1979-01-01

    The results of two radioimmunoassays (RIA, precipitating technique), of a homogenous (EMIT) and a heterogenous (ELISA) enzyme immunoassay (EIA) for ascertaining the amounts of digoxin showed a good correlation in precision and a reasonably AK satisfying correlation in the recovery. However, there was a clear discrepancy in the amounts of digoxin concentrate in the serum of patients. Only the RIA of Abbott and the EIA of Boehringer showed no significant differences. Particularly noticeable was the tendency towards lower values in the EMIT-technique as well as its liability to unspecific serum changes (lipaemia etc.), which often made the detection of digoxin impossible. The routine use of this technique appears problematic. The need for establishing one's own laboratory and test-specific therapeutical range is pointed out. (orig.) [de

  10. Establishment of a Simple and Convenient Method for Folic Acid Enzyme Chemiluminescence Immunoassay

    Science.gov (United States)

    Liu, Ting; Zeng, Ling; Yu, Zhengwei; Yang, Yanfei; Qu, Yunhuan

    2018-01-01

    The enzyme chemiluminescence new immunoassay for folic acid (FA) was established by competition model. Add FA samples to a microtiter plate precoated with the goat anti-mouse IgG firstly, then add enzyme abled FA and FA monoclonal antibody (McAb). The values of CLIA were measured to reflect the quantity of FA. The limit of detection(LOD) of assay is 0.37ng/mL. The assay shows good correlation during 1∼30 ng/mL with correlation coefficient 0.9976. The intra- and inter-assay coefficients of variation are 4.8 % ∼ 7.3 % and 6.1 % ∼ 12.2 %, respectively. The recovery of folic acid in serum is 90.4 %∼113.2 %. Compared with determine value clinically in chemiluminescence immunoassay kit from Roche company, the correlative equation is y = 0.9689x + 0.0228, and correlation coefficient is 0.9780. Various components and kit overall show good stabilities. This method is simple and convenient, and has low LOD value. The method has overcome the shortcomings of the present references. It is easy to apply and has broad clinical application prospect. It lays an experimental foundation for the preparation of Mc Ab against folic acid and the development of domestic kit.

  11. Human chorionic gonadotropin detection in cerebrospinal fluid of patients with a germinoma and its prognostic significance: assessment by using a highly sensitive enzyme immunoassay.

    Science.gov (United States)

    Fukuoka, Kohei; Yanagisawa, Takaaki; Suzuki, Tomonari; Shirahata, Mitsuaki; Adachi, Jun-Ichi; Mishima, Kazuhiko; Fujimaki, Takamitsu; Katakami, Hideki; Matsutani, Masao; Nishikawa, Ryo

    2016-11-01

    OBJECTIVE Human chorionic gonadotropin (HCG) can be detected in a certain population of patients with a germinoma, but the frequency of germinoma HCG secretion and the prognostic value of HCG in the CSF are unknown. METHODS The authors measured HCG levels in sera and CSF in patients with a histologically confirmed germinoma by using a highly sensitive assay known as an immune complex transfer enzyme immunoassay (EIA), which is more than 100 times as sensitive as the conventional method, and they analyzed the correlation between HCG levels and the prognoses of patients with a germinoma. RESULTS HCG levels in sera and CSF of 35 patients with a germinoma were examined with the immune complex transfer EIA. The median CSF HCG levels in patients with a germinoma during the pretreatment and posttreatment evaluations were 192.5 pg/ml (range 1.2-13,116.5 pg/ml) and 18.7 pg/ml (1.2-283.9 pg/ml), respectively. Before treatment, the CSF HCG level was greater than the cutoff value in 85.7% of the patients with a germinoma. The authors compared survival rates among the patients by using a CSF HCG cutoff level of 1000 pg/ml, and the difference was statistically significant between the groups (p = 0.029, log-rank test). CONCLUSIONS Results of this study demonstrate that most germinomas secrete HCG. Patients with a germinoma that secretes higher amounts of HCG in their CSF experienced recurrence more frequently than those with lower CSF HCG levels.

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

    DEFF Research Database (Denmark)

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

    2014-01-01

    using IFA and automated EIA techniques. The IFA results generated by two independent laboratories were compared with the EIA results from antibodies against double-stranded DNA (dsDNA), from ANA screening, and from tests of the seven included subantigens. The final IFA and EIA results for 386 unique......, with Cohen's kappa value of 0.30 (95% confidence interval (CI) = 0.14-0.46), which decreased to 0.23 (95% CI = 0.06-0.40) when the results for dsDNA were omitted. The EIA method was less reliable for assessing nuclear and speckled reactivity patterns, whereas the IFA method presented difficulties detecting...... dsDNA and Ro activity. The automated EIA method was performed in a similar way to the conventional IFA method using HEp-2 cells; thus, automated EIA may be used as a screening test....

  13. Validation of a Commercially Available Enzyme ImmunoAssay for the Determination of Oxytocin in Plasma Samples from Seven Domestic Animal Species

    Directory of Open Access Journals (Sweden)

    Cecile Bienboire-Frosini

    2017-09-01

    Full Text Available The neurohormone oxytocin (OT has a broad range of behavioral effects in mammals. It modulates a multitude of social behaviors, e.g., affiliative and sexual interactions. Consequently, the OT role in various animal species is increasingly explored. However, several issues have been raised regarding the peripheral OT measurement. Indeed, various methods have been described, leading to assay discrepancies and inconsistent results. This highlights the need for a recognized and reliable method to measure peripheral OT. Our aim was to validate a method combining a pre-extraction step, previously demonstrated as essential by several authors, and a commercially available enzyme immunoassay (EIA for OT measurement, using plasma from seven domestic species (cat, dog, horse, cow, pig, sheep, and goat. The Oxytocin EIA kit (EnzoLifeSciences was used to assay the solid-phase extracted samples following the manufacturer's instructions with slight modifications. For all species except dogs and cats, concentration factors were applied to work above the kit's sensitivity (15 pg/ml. To validate the method, the following performance characteristics were evaluated using Validation Samples (VS at various concentrations in each species: extraction efficiency via spiking tests and intra- and inter-assay precision, allowing for the calculation of total errors. Parallelism studies to assess matrix effects could not be performed because of too low basal concentrations. Quantification ranges and associated precision profiles were established to account for the various OT plasma concentrations in each species. According to guidelines for bioanalytical validation of immunoassays, the measurements were sufficiently precise and accurate in each species to achieve a total error ≤30% in each VS sample. In each species, the inter-assay precision after 3 runs was acceptable, except in low concentration samples. The linearity under dilution of dogs and cats' samples was

  14. Validation of a Commercially Available Enzyme ImmunoAssay for the Determination of Oxytocin in Plasma Samples from Seven Domestic Animal Species.

    Science.gov (United States)

    Bienboire-Frosini, Cecile; Chabaud, Camille; Cozzi, Alessandro; Codecasa, Elisa; Pageat, Patrick

    2017-01-01

    The neurohormone oxytocin (OT) has a broad range of behavioral effects in mammals. It modulates a multitude of social behaviors, e.g., affiliative and sexual interactions. Consequently, the OT role in various animal species is increasingly explored. However, several issues have been raised regarding the peripheral OT measurement. Indeed, various methods have been described, leading to assay discrepancies and inconsistent results. This highlights the need for a recognized and reliable method to measure peripheral OT. Our aim was to validate a method combining a pre-extraction step, previously demonstrated as essential by several authors, and a commercially available enzyme immunoassay (EIA) for OT measurement, using plasma from seven domestic species (cat, dog, horse, cow, pig, sheep, and goat). The Oxytocin EIA kit (EnzoLifeSciences) was used to assay the solid-phase extracted samples following the manufacturer's instructions with slight modifications. For all species except dogs and cats, concentration factors were applied to work above the kit's sensitivity (15 pg/ml). To validate the method, the following performance characteristics were evaluated using Validation Samples (VS) at various concentrations in each species: extraction efficiency via spiking tests and intra- and inter-assay precision, allowing for the calculation of total errors. Parallelism studies to assess matrix effects could not be performed because of too low basal concentrations. Quantification ranges and associated precision profiles were established to account for the various OT plasma concentrations in each species. According to guidelines for bioanalytical validation of immunoassays, the measurements were sufficiently precise and accurate in each species to achieve a total error ≤30% in each VS sample. In each species, the inter-assay precision after 3 runs was acceptable, except in low concentration samples. The linearity under dilution of dogs and cats' samples was verified. Although

  15. Detection of Pesticides and Pesticide Metabolites Using the Cross Reactivity of Enzyme Immunoassays

    Science.gov (United States)

    Thurman, E.M.; Aga, D.S.

    2001-01-01

    Enzyme immunoassay is an important environmental analysis method that may be used to identify many pesticide analytes in water samples. Because of similarities in chemical structure between various members of a pesticide class, there often may be an unwanted response that is characterized by a percentage of cross reactivity. Also, there may be cross reactivity caused by degradation products of the target analyte that may be present in the sample. In this paper, the concept of cross reactivity caused by degradation products or by nontarget analytes is explored as a tool for identification of metabolites or structurally similar compounds not previously known to be present in water samples. Two examples are examined in this paper from various water quality studies. They are alachlor and its metabolite, alachlor ethane sulfonic acid, and atrazine and its class members, prometryn and propazine. A method for using cross reactivity for the detection of these compounds is explained in this paper.

  16. Quantification of low levels of rainbow trout immunoglobulin by enzyme immunoassay using two monoclonal antibodies.

    Science.gov (United States)

    Sánchez, C; Babin, M; Tomillo, J; Ubeira, F M; Domínguez, J

    1993-02-01

    An enzyme immunoassay has been developed for quantitation of low levels of trout immunoglobulin (Ig). This assay uses two monoclonal antibodies, one as capture antibody and the other as detector, directed against two non-overlapping epitopes on the heavy chains of trout Ig. The assay shows high reproducibility and can detect 0.12 micrograms trout Ig ml-1. Coefficients of intra- and interassay variation ranged from 3.8 to 7.1% and from 7.9 to 17.4%, respectively. Analysis of 37 healthy trout showed increasing serum Ig concentration with size. The mean Ig concentration was 0.67 mg ml-1 for trout of about 20 g and 9.1 mg ml-1 for trout weighing more than 125 g.

  17. Evaluation of the concomitant use of two different EIA tests for HIV screening in blood banks.

    Science.gov (United States)

    Otani, Marcia M; Salles, Nanci A; Barreto, Angela M E; Barreto, Claudia C; Chamone, Dalton F; Sabino, Ester C

    2003-01-01

    In 1998, the Brazilian Ministry of Health made it mandatory for all blood banks in the country to screen donated blood for human immunodeficiency virus (HIV) concomitantly using two different enzyme immunoassay (EIA) tests. Concerned with the best use of available resources, our objective with this study was to evaluate the usefulness of conducting two EIA screening tests instead of just one. We analyzed data from 1999 through 2001 obtained by testing 698 191 units of donated blood using two EIA HIV screening tests concomitantly at the Pro-Blood Foundation/Blood Center of São Paulo (Fundação Pró-Sangue/Hemocentro de São Paulo), which is a major blood center in the city of São Paulo, Brazil. All samples reactive in at least one of the two EIA tests were submitted for confirmation by a Western blot (WB) test, and the persons who had donated those samples were also asked to return and provide a follow-up sample. Out of the 698 191 blood units that were donated, 2 718 of them (0.4%) had to be discarded because they were reactive to at least one of the EIA tests. There were two WB-positive donation samples that were reactive in only one HIV EIA screening test. On their follow-up samples, both donors tested WB-negative. These cases were considered false positive results at screening. Of the 2 718 donors who were asked to return and provide a follow-up sample, 1 576 of them (58%) did so. From these 1 576 persons, we found that there were two individuals who had been reactive to only one of the two EIA screening tests and who had also been negative on the WB at screening but who were fully seroconverted on the follow-up sample. We thus estimated that, in comparison to the use of a single EIA screening test, the use of two EIA screening tests would detect only one extra sample out of 410 700 units of blood. Our data do not support the use of two different, concomitant EIA screening tests for HIV. The great majority of HIV-positive donors have already seroconverted and

  18. Detection of hidden hazelnut protein in food by IgY-based indirect competitive enzyme-immunoassay

    NARCIS (Netherlands)

    Baumgartner, S.; Bremer, M.G.E.G.; Kemmers - Voncken, A.E.M.; Smits, N.G.E.; Haasnoot, W.; Banks, J.; Reece, P.; Danks, C.; Tomkies, V.; Immer, U.; Schmitt, K.; Krska, R.

    2004-01-01

    The development of an indirect competitive enzyme-immunoassay for the detection of hidden hazelnut protein in complex food matrices is described. A sensitive and selective polyclonal antibody was raised by immunisation of laying hens with protein extracts from roasted hazelnuts. In contrast to

  19. Quantification of patient specific assay interference in different formats of enzyme linked immunoassays for therapeutic monoclonal antibodies

    NARCIS (Netherlands)

    Grebenchtchikov, N.J.; Geurts-Moespot, A.; Heijmen, L.; Laarhoven, H.W.M. van; Herpen, C.M.L. van; Thijs, A.M.J.; Span, P.N.; Sweep, F.C.

    2014-01-01

    BackgroundThe use of therapeutic monoclonal antibodies for clinical purposes has significantly increased in recent years, and so has the need to monitor antibody concentrations. This may be achieved using the well-established enzyme linked immunoassay (ELISA) methods; however, these assays are

  20. Quantification of patient-specific assay interference in different formats of enzyme-linked immunoassays for therapeutic monoclonal antibodies

    NARCIS (Netherlands)

    Grebenchtchikov, Nicolai; Geurts-Moespot, Anneke J.; Heijmen, Linda; van Laarhoven, Hanneke W. M.; van Herpen, Carla M. L.; Thijs, Annemarie M. J.; Span, Paul N.; Sweep, Fred C. G. J.

    2014-01-01

    The use of therapeutic monoclonal antibodies for clinical purposes has significantly increased in recent years, and so has the need to monitor antibody concentrations. This may be achieved using the well-established enzyme linked immunoassay (ELISA) methods; however, these assays are subject to a

  1. Immunoassays of fungal laccases for screening of natural enzymes and control of recombinant enzyme production.

    Science.gov (United States)

    Loginov, Dmitry S; Vavilova, Ekaterina A; Savinova, Оlga S; Abyanova, Alfia R; Chulkin, Andrey M; Vasina, Daria V; Zherdev, Anatoly V; Koroleva, Olga V

    2014-01-01

    Because of the wide application of laccases in different biotechnological processes and intense studies of the enzymes from different sources, the development of efficient techniques for monitoring laccase level is a task of significant importance. Enzyme-linked immunosorbent assay (ELISA) and Western blotting techniques were developed to control total content and isoform composition of laccases, including their recombinant preparations. Because glycosylated and nonglycosylated forms have different structures and sets of epitopes, two kinds of polyclonal antibodies were obtained and applied. The first antibody recognized the native (glycosylated) laccase purified from Trametes hirsuta and the second one reacted with recombinant (nonglycosylated) laccase expressed in Escherichia coli. Titers of the antibodies were analyzed by indirect ELISA with laccases isolated from several strains of basidiomycetes. The obtained cross-reactivity data for both antibodies demonstrated a correspondence with sequence homology of the laccases. The antibodies raised against recombinant (nonglycosylated) laccase had higher titers and thus were preferable for screening of recombinant laccase in cultural media. Thus, optimal antibody preparations were selected for screening of laccase-producing strains, and the control of recombinant enzymes and the efficiency of their use in immunochemical control of laccase levels were confirmed. © 2013 International Union of Biochemistry and Molecular Biology, Inc.

  2. Androgen and androgen metabolite levels in serum and urine of East African chimpanzees (Pan troglodytes schweinfurthii): comparison of EIA and LC-MS analyses.

    Science.gov (United States)

    Preis, Anna; Mugisha, Lawrence; Hauser, Barbara; Weltring, Anja; Deschner, Tobias

    2011-12-01

    The primary male androgen testosterone (T) is often used as an endocrinological marker to investigate androgen-behaviour interactions in males. In chimpanzees and bonobos, studies investigating the relationship between T levels and dominance rank or aggressive behaviour have revealed contradictory results. The immunoassays used in these studies were originally developed for the measurement of steroids in serum. Their application to non-invasively collected samples, however, can lead to methodological problems due to cross-reacting metabolites, which might occur in urine or faeces but not in blood. The overall aim of this study, therefore, is to clarify whether a T enzyme immunoassay (EIA) is an applicable method to monitor testicular function in adult male chimpanzees. To estimate the impact of cross-reacting androgens on the used T EIA, we compared the results of an EIA measurement with a set of androgen metabolite levels measured by LC-MS. In urine from male chimpanzees, cross-reactivities appear to exist mainly with T and its exclusive metabolites, 5α-dihydrotestosterone (5α-DHT) and 5α-androstanediol (androstanediol). Both urinary and serum T levels of male chimpanzees were significantly higher than female T levels when measured with the T EIA, indicating a reliable measurement of testicular androgens and their exclusive metabolites with the used EIA. In urine from female chimpanzees, the comparison between LC-MS and T EIA results indicated a higher impact of cross-reactions with adrenal androgen metabolites. Therefore, the investigation of urinary T levels in female chimpanzees with a T EIA seems to be problematic. Overall our results show that a T EIA can be a reliable method to monitor testicular function in male chimpanzee urine and that LC-MS is a valuable tool for the validation of immunoassays. Copyright © 2011 Elsevier Inc. All rights reserved.

  3. Development of an efficient signal amplification strategy for label-free enzyme immunoassay using two site-specific biotinylated recombinant proteins

    Energy Technology Data Exchange (ETDEWEB)

    Tang, Jin-Bao [School of Pharmacy, Weifang Medical University, Weifang 261053 (China); Tang, Ying [Affiliated Hospital of Weifang Medical University, Weifang 261041 (China); Yang, Hong-Ming, E-mail: yanghongming2006@sohu.com [School of Pharmacy, Weifang Medical University, Weifang 261053 (China)

    2015-02-15

    Highlights: • An efficient signal amplification strategy for label-free EIA is proposed. • Divalent biotinylated AP and monovalent biotinylated ZZ were prepared via Avitag–BirA system. • The above site-specific biotinylated fusion proteins form complex via SA–biotin interaction. • The mechanism relies on the ZZ–Avi-B/SA/AP–(Avi-B){sub 2} complex. • The analytical signals are enhanced (32-fold) by the proposed strategy. - Abstract: Constructing a recombinant protein between a reporter enzyme and a detector protein to produce a homogeneous immunological reagent is advantageous over random chemical conjugation. However, the approach hardly recombines multiple enzymes in a difunctional fusion protein, which results in insufficient amplification of the enzymatic signal, thereby limiting its application in further enhancement of analytical signal. In this study, two site-specific biotinylated recombinant proteins, namely, divalent biotinylated alkaline phosphatase (AP) and monovalent biotinylated ZZ domain, were produced by employing the Avitag–BirA system. Through the high streptavidin (SA)–biotin interaction, the divalent biotinylated APs were clustered in the SA–biotin complex and then incorporated with the biotinylated ZZ. This incorporation results in the formation of a functional macromolecule that involves numerous APs, thereby enhancing the enzymatic signal, and in the production of several ZZ molecules for the interaction with immunoglobulin G (IgG) antibody. The advantage of this signal amplification strategy is demonstrated through ELISA, in which the analytical signal was substantially enhanced, with a 32-fold increase in the detection sensitivity compared with the ZZ–AP fusion protein approach. The proposed immunoassay without chemical modification can be an alternative strategy to enhance the analytical signals in various applications involving immunosensors and diagnostic chips, given that the label-free IgG antibody is suitable

  4. The clinical application of fluorescent-enzyme immunoassay to detect human thyroid peroxidase autoantibody quantitatively

    International Nuclear Information System (INIS)

    Chai Jinyan; Fang Peihua; Li Ning; Zhang Yanli

    2010-01-01

    Objective: To establish a fluorescent-enzyme immunoassay (FEIA) with recombinant human thyroid peroxidase (hTPO) as the antigen. Methods: Sera of 326 healthy people, 119 cases of Hashimoto's thyroiditis (HT), 116 cases of Graves disease (untreated), 28 cases of nodular goiter, 10 ca-ses of subacute thyroiditis and 6 cases of simple goiter were measured by the FEIA with recombinant hTPO as the antigen. Rrank sum test and χ 2 -test were used in inter-groups. Results: (1) Concentration above 4000 U/L was considered to be positive. (2) The intra-assay coefficient of variation (CV) and the inter-as-say CV were 4.59% ∼ 6.52% and 17.37% ∼ 17.45%. (3) The values measured by the FEIA were positively correlated with the values measured by hTPO antibody (hTPOAb) commercial kit (r=0.80, P 2 = 53.45, 39.30, 15.41 and 21.74, all P < 0.01). Conclusions: The method can be applied in the serum measurement of thyroid disease. HT presented the highest positive rate, therefore, the FEIA was an effective method in diagnosing HT. (authors)

  5. [Use of monoclonal antibodies against horse immunoglobulin in an enzyme immunoassay of bacterial toxins and anatoxins].

    Science.gov (United States)

    Burkin, M A; Gal'vidis, I A; Iakovleva, I V; Sviridov, V V

    2007-01-01

    Immunization of BALB/c mice by horse antiserum against diphtheria made it possible to obtain IgG1 monoclonal antibodies (MoAbs) 2B7E4 specific for light chains of horse immunoglobulin (Ig). Unlike commercial preparations of anti-horse immunoglobulin antibodies, which are specific for the whole Ig molecule or its Fc-fragment, the peroxidase (HRP) conjugate of the MoAb, 2B7E4-HRP did not interact with human, mouse, rabbit, and sheep Igs, or horse albumin. The conjugate obtained was used with MoAbs against bacterial toxins and commercial horse anatoxins, as a universal reagent in sandwich enzyme immunoassay (ELISA) for bacterial toxins and anatoxins. The detection sensitivity of diphtheria toxin/anatoxin equaled 0.0005 Lf/ml; tetanus toxin and anatoxin were detected with sensitivities of 20 LD50/ml and 0.005 UI/ml, respectively. A similar sandwich ELISA for botulinum anatoxins (group measurement) allowed types A, B, and E to be detected at 0.02, 0.002, and 0.001 UI/ml, respectively; selective measurement was only possible in the case of type E anatoxin (0.001 UI/ml).

  6. Enzyme immunoassays for the diagnosis of bovine brucellosis. Trial in Latin America

    International Nuclear Information System (INIS)

    Gall, D.; Nielsen, K.; Colling, A.; Marino, O.; Moreno, E.; Perez, B.; Samartino, L.

    1998-01-01

    The results of a field trail conducted in Latin America with two indirect (IELISA) and two competitive (CELISA) enzyme immunoassays for the detection of bovine antibody to Brucella abortus are reported. One of the CELISA formats performed most accurately. The relative sensitivity of this assay was 97.47%, the relative specificity for unexposed cattle was 98.32% and the specificity in cattle vaccinated with B. abortus strain 19 was 96.51%. The same assay format under Canadian conditions had a sensitivity of 100%, a specificity of 99.90% and a specificity of 97.7% in a strain 19 vaccinated population. Overall, the CELISA performed as expected and the results were not dissimilar to the results obtained in the Canadian study thus providing further evidence that this CELISA can in many instances differentiate infected cattle from those that are vaccinated or infected with a cross-reacting organism while still giving very low false positive or false negative results. (author)

  7. Nanobody-based enzyme immunoassay for ochratoxin A in cereal with high resistance to matrix interference.

    Science.gov (United States)

    Liu, Xing; Tang, Zongwen; Duan, Zhenhua; He, Zhenyun; Shu, Mei; Wang, Xianxian; Gee, Shirley J; Hammock, Bruce D; Xu, Yang

    2017-03-01

    A sensitive indirect competitive nanobody-based enzyme linked immunosorbent assay (Nb-ELISA) for ochratoxin A (OTA) with high resistance to cereal matrix interference was developed. Nanobodies against OTA (Nb15, Nb28, Nb32, Nb36) were expressed in E. coli cells and their thermal stabilities were compared with that of an OTA-specific monoclonal antibody 6H8. All nanobodies could still retain their antigen-binding activity after exposure to temperature 95°C for 5min or to 90°C for 75min. Nb28 that exhibited the highest sensitivity in ELISA was selected for further research. An indirect competitive ELISA based on Nb28 was developed for OTA, with an IC 50 of 0.64ng/mL and a linear range (IC 20 -IC 80 ) of 0.27-1.47ng/mL. Cereal samples were analyzed following a 2.5 fold dilution of sample extracts, showing the good resistance to matrix interference of the Nb-ELSIA. The recovery of spiked cereal samples (rice, oats, barley) ranged from 80% to 105% and the Nb-ELISA results of OTA content in naturally contamined samples were in good agreement with those determined by a commercial ELISA kit. The results indicated the reliablity of nanobody as a promising immunoassay reagent for detection of mycotoxins in food matrix and its potential in biosensor development. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. Multicentric Evaluation of New Commercial Enzyme Immunoassays for the Detection of Immunoglobulin M and Total Antibodies against Hepatitis A Virus▿

    Science.gov (United States)

    Arcangeletti, M. C.; Dussaix, E.; Ferraglia, F.; Roque-Afonso, A. M.; Graube, A.; Chezzi, C.

    2011-01-01

    A multicentric clinical study was conducted on representative sera from 1,738 European and U.S. subjects for the evaluation of new anti-hepatitis A virus enzyme immunoassays from Bio-Rad Laboratories. Comparison with reference DiaSorin S.p.A. tests confirmed the good performance of Bio-Rad assays (99.85% and 99.47% overall agreement in detecting total antibodies and IgM, respectively). PMID:21653739

  9. Detection of virus-specific intrathecally synthesised immunoglobulin G with a fully automated enzyme immunoassay system

    Directory of Open Access Journals (Sweden)

    Weissbrich Benedikt

    2007-05-01

    Full Text Available Abstract Background The determination of virus-specific immunoglobulin G (IgG antibodies in cerebrospinal fluid (CSF is useful for the diagnosis of virus associated diseases of the central nervous system (CNS and for the detection of a polyspecific intrathecal immune response in patients with multiple sclerosis. Quantification of virus-specific IgG in the CSF is frequently performed by calculation of a virus-specific antibody index (AI. Determination of the AI is a demanding and labour-intensive technique and therefore automation is desirable. We evaluated the precision and the diagnostic value of a fully automated enzyme immunoassay for the detection of virus-specific IgG in serum and CSF using the analyser BEP2000 (Dade Behring. Methods The AI for measles, rubella, varicella-zoster, and herpes simplex virus IgG was determined from pairs of serum and CSF samples of patients with viral CNS infections, multiple sclerosis and of control patients. CSF and serum samples were tested simultaneously with reference to a standard curve. Starting dilutions were 1:6 and 1:36 for CSF and 1:1386 and 1:8316 for serum samples. Results The interassay coefficient of variation was below 10% for all parameters tested. There was good agreement between AIs obtained with the BEP2000 and AIs derived from the semi-automated reference method. Conclusion Determination of virus-specific IgG in serum-CSF-pairs for calculation of AI has been successfully automated on the BEP2000. Current limitations of the assay layout imposed by the analyser software should be solved in future versions to offer more convenience in comparison to manual or semi-automated methods.

  10. Geography, Planning and EIA.

    Science.gov (United States)

    Morgan, R . K.

    1987-01-01

    Defines Environmental Impact Assessment (EIA) as a major tool for environmental management throughout the world. Provides background to the EIA concept and describes how it might be used within secondary school geography as an integrative teaching device. Special emphasis is given to the new EIA/Seventh-Form syllabus currently being used in New…

  11. NON-RADIOMETRIC IMMUNOASSAYS [FLUOROIMMUNOASSAY (FIA AND FLUOROMETRIC ENZYME IMMUNOASSAY (FEIA] WITH RADIOIMMUNOASSAY (RIA FOR EVALUATION OF ADRENAL FUNCTION IN NORMAL AND HYPERCORTISOLEMIC DOGS

    Directory of Open Access Journals (Sweden)

    Jericó Márcia Marques

    2002-01-01

    Full Text Available Non-radiometric immunoassays offer many advantages over radiometric assays, such as higher stability of kit compounds and absence of potential hazardous effects for users and environment. The comparison of cortisol measurements by fluoroimmunoassay (FIA and fluorometric enzyme immunoassay (FEIA with radioimmunoassay (RIA in adrenal function evaluation of normal (n=50 and hypercortisolemic dogs (n=12 was proposed. Serum concentrations of cortisol were measured in basal conditions and 8 hours after dexamethasone (DEX suppression (0.01mg/kg/IV. All our reference values were based on the 5th and 95th percentile. The values for basal cortisol of healthy dogs were 0.20 to 2.35mug/d for FIA, 0.30 to 5.39mug/d for FEIA, and 0.65 to 4.64mug/d for RIA. After DEX suppression the values were , and for FIA, FEIA and RIA, respectively. In hypercortisolemic dogs, the values of cortisol (mean ± SD in basal and post-DEX conditions were 2.71 + 0.41mug/d and 1.73 + 1.15mug/d for FIA, 7.05 + 2.85mug/d and 4.93 + 2.26mug/d for FEIA, and 4.80 + 1.43mug/d and 3.52 + 1.08mug/d for RIA. Statistically significant differences (p<0.05 between the normal and the hypercortisolemic groups (Kruskal-Wallis test were observed in the three methods, and between basal and post-DEX values (Wilcoxon test using RIA and FEIA methods but not with FIA. Cortisol determinations by FEIA and RIA methods at DEX suppression test showed 100% of sensitivity and specificity for the diagnosis of hyperadrenocorticism in dogs. The results demonstrate that serum cortisol concentrations measurements by FEIA is a suitable alternative to the traditional RIA method for adrenal function evaluation in dogs.

  12. Towards rationalisation of antivenom use in funnel-web spider envenoming: enzyme immunoassays for venom concentrations.

    Science.gov (United States)

    Miller, Mark; O'Leary, Margaret A; Isbister, Geoffrey K

    2016-03-01

    Funnel-web spider (Atrax and Hadronyche spp.) envenoming is rare but causes severe neuromuscular, autonomic, and cardiac effects. A rabbit-derived IgG antivenom is available, but venom detection in patients has not been reported. To use serial venom and antivenom concentrations to better define envenoming and antivenom effectiveness. Serum was collected from nine patients with suspected funnel-web spider bites and clinical effects were recorded. Venom-specific enzyme immunoassays were developed to measure funnel-web spider venom and antivenom concentrations. Goat anti-rabbit whole serum was coupled to UltraLink resin and added to samples to remove bound venom and measure free venom. Antivenom efficacy was defined as antivenom binding all free venom and antivenom effectiveness as resolution of clinical features. Venom was detectable in samples from six of nine patients. In three patients without venom detected, there were only moderate effects, which did not completely respond to antivenom in all cases and no spider was identified. In five of six cases, a male Atrax spp. (Sydney funnel-web) spider was identified. Three patients had moderate envenoming which responded to antivenom. Three patients had severe envenoming and developed catecholamine-induced myocarditis and acute pulmonary oedema. Although cholinergic and non-specific clinical features appeared to respond to antivenom, myocarditis and pulmonary oedema lasted 2-4 days. Median venom concentration pre-antivenom in five patients with samples was 5.6 ng/ml (3-35 ng/ml), and immediately post-antivenom decreased to a median of 0 ng/ml (0-1.8 ng/ml). Post-antivenom venom concentrations decreased when bound venom was removed; median, 0 ng/ml (0-0.9 ng/ml), indicating that most venom detected post-antivenom was bound. There was recurrence of venom and clinical features in one patient when a pressure bandage was removed. Detection of venom in suspected funnel-web spider bites identified definite cases with

  13. Magnetic-particle-based, ultrasensitive chemiluminescence enzyme immunoassay for free prostate-specific antigen

    International Nuclear Information System (INIS)

    Liu, Ruping; Wang, Cheng; Jiang, Quan; Zhang, Wei; Yue, Zhao; Liu, Guohua

    2013-01-01

    Graphical abstract: -- Highlights: •A low-cost and rapid assay for f-PSA in serum was developed using MMP-based CLEIA. •f-PSA detection: 0.1–30 ng mL −1 concentration range; 0.1 ng mL −1 detection limit. •The proposed method showed high sensitivity, good reproducibility and stability. •The strategy showed great potential in the fabrication of MMP-based f-PSA test kits. -- Abstract: We report a magnetic-particle (MMP)-based chemiluminescence enzyme immunoassay (CLEIA) for free prostate-specific antigen (f-PSA) in human serum. In this method, the f-PSA is sandwiched between the anti-PSA antibody coated MMPs and alkaline phosphatase (ALP)-labeled anti-f-PSA antibody. The signal produced by the emitted photons from the chemiluminescent substrate (4-methoxy-4-(3-phosphatephenyl)-spiro-(1,2-dioxetane-3,2′-adamantane)) is directly proportional to the amount of f-PSA in a sample. The present MMP-based assay can detect f-PSA in the range of 0.1–30 ng mL −1 with the detection limit of 0.1 ng mL −1 . The linear detection range could match the concentration range within the “diagnostic gray zone” of serum f-PSA levels (4–10 ng mL −1 ). The detection limit was sufficient for measuring clinically relevant f-PSA levels (>4 ng mL −1 ). Furthermore, the method was highly selective; it was unaffected by cross-reaction with human glandular kallikrein-2, a kallikrein-like serine protease that is 80% similar to f-PSA. The proposed method was finally applied to determine f-PSA in 40 samples of human sera. Results obtained using the method showed high correlation with those obtained using a commercially available microplate CLEIA kit (correlation coefficient, 0.9821). This strategy shows great potential application in the fabrication of diagnostic kits for determining f-PSA in serum

  14. Magnetic-particle-based, ultrasensitive chemiluminescence enzyme immunoassay for free prostate-specific antigen

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Ruping [College of Electronic Information and Optical Engineering, Nankai University, Tianjin 300071 (China); Wang, Cheng [College of Electronic Information and Optical Engineering, Nankai University, Tianjin 300071 (China); Department of Mechanical Engineering, Columbia University, New York 10027 (United States); Jiang, Quan [Institute of Material Science and Engineering, Wuhan University of Technology, Wuhan 430070 (China); Zhang, Wei; Yue, Zhao [College of Electronic Information and Optical Engineering, Nankai University, Tianjin 300071 (China); Liu, Guohua, E-mail: liugh@nankai.edu.cn [College of Electronic Information and Optical Engineering, Nankai University, Tianjin 300071 (China)

    2013-11-01

    Graphical abstract: -- Highlights: •A low-cost and rapid assay for f-PSA in serum was developed using MMP-based CLEIA. •f-PSA detection: 0.1–30 ng mL{sup −1} concentration range; 0.1 ng mL{sup −1} detection limit. •The proposed method showed high sensitivity, good reproducibility and stability. •The strategy showed great potential in the fabrication of MMP-based f-PSA test kits. -- Abstract: We report a magnetic-particle (MMP)-based chemiluminescence enzyme immunoassay (CLEIA) for free prostate-specific antigen (f-PSA) in human serum. In this method, the f-PSA is sandwiched between the anti-PSA antibody coated MMPs and alkaline phosphatase (ALP)-labeled anti-f-PSA antibody. The signal produced by the emitted photons from the chemiluminescent substrate (4-methoxy-4-(3-phosphatephenyl)-spiro-(1,2-dioxetane-3,2′-adamantane)) is directly proportional to the amount of f-PSA in a sample. The present MMP-based assay can detect f-PSA in the range of 0.1–30 ng mL{sup −1} with the detection limit of 0.1 ng mL{sup −1}. The linear detection range could match the concentration range within the “diagnostic gray zone” of serum f-PSA levels (4–10 ng mL{sup −1}). The detection limit was sufficient for measuring clinically relevant f-PSA levels (>4 ng mL{sup −1}). Furthermore, the method was highly selective; it was unaffected by cross-reaction with human glandular kallikrein-2, a kallikrein-like serine protease that is 80% similar to f-PSA. The proposed method was finally applied to determine f-PSA in 40 samples of human sera. Results obtained using the method showed high correlation with those obtained using a commercially available microplate CLEIA kit (correlation coefficient, 0.9821). This strategy shows great potential application in the fabrication of diagnostic kits for determining f-PSA in serum.

  15. Measurement of Urinary Growth Hormone (GH) Levels by Highly Sensitive Enzyme Immunoassay as a Potential Screening Test for GH Secretion

    OpenAIRE

    Sohmiya, Motoi; Kato, Yuzuru

    1992-01-01

    We set up a highly sensitive enzyme immunoassay of human growth hormone (hGH) using anti-hGH rabbit Fab'-peroxidase conjugate. IgG was obtained from anti-hGH rabbit serum with salting-out and diethylaminoethyl cellulose. IgG was digested by porcine gastric mucosa pepsin to F (ab')_2 which was reduced to Fab'. Fab' was conjugated with peroxidase by maleimide method. The minimum detectable amount of hGH was 0.3pg/ml using 100μl of dialyzed urine sample without any concentrating procedure. Urina...

  16. The clinical value of enzyme-multiplied immunoassay technique monitoring the plasma concentrations of cyclosporine A after renal transplantation

    Directory of Open Access Journals (Sweden)

    Xiao-Hui Luo

    2011-05-01

    Full Text Available The feasibility and the clinical value of the enzyme-multiplied immunoassay technique (EMIT monitoring of blood concentrations of cyclosporine A (CsA in patients treated with CsA were investigated after kidney transplantation. The validation method was performed to the EMIT determination of CsA blood concentration, the CsA whole blood ‘trough concentrations (C0 of patients in different time periods after renal transplantation were monitored, and combined with the clinical complications, the statistical results were analyzed and compared. EMIT was precise, accurate and stable, also with a high quality control. The mean postoperative blood concentration of CsA was as follows: 12 months, (185.6 ± 28.1ng/mL. The toxic reaction rate of CsA blood concentration within the recommended therapeutic concentration was 14. 1%, significantly lower than that of the none-recommended dose group (37.2% (P < 0.05; the transplantation rejection rate was 4.4%, significantly lower than that of the none-recommended dose group (22.5% (P < 0.05. Using EMIT to monitor the blood concentration of CsA as the routine laboratory method is feasible, and is able to reduce the CsA toxicity and rejection significantly, leading to achieving the desired therapeutic effect. Keywords: enzyme-multiplied immunoassay technique, renal transplantation, cyclosporin A, blood concentration monitoring

  17. The influence of intravenous canrenoate on the determination of digoxin in serum by radio- and enzyme-immunoassay

    International Nuclear Information System (INIS)

    Rietbrock, N.; Lichey, J.; Borner, K.; Freie Univ. Berlin

    1979-01-01

    Ten patients were kept on a constant maintenance dose of digoxin. During a baseline period of 6 days, blood samples were taken daily for analysis of digoxin in serum. On the 6th day the maintenance dose of digoxin was withheld and a single intravenous dose of 200mg potassium-canrenoate (AldactoneR) was administered to all patients. Digoxin in serum was determined by a classical radioimmunoassay with 125 I-digoxin and solid phase technique (RIA-NEN) and partly by a heterogenous enzyme-immunoassay (EnzymunR-Digoxin, Boehringer, Mannheim). Results of the radioimmunoassay indicated a rise of apparent serum digoxin levels with an average maximum of 201% of the mean baseline value 30 min after injection of canrenoate and a gradual return to the baseline value within 6 to 10 hours. Contrary to the radioimmunoassay there was no interference when using the enzyme-immunoassay in a subgroup of identical serum samples: serum digoxin levels remained constant throughout the test. Interference of determinations of digoxin in serum by spironolactone and its metabolites appear to be related to two factors: 1. The mode of administration and the amount of interfering drug, 2. the specifity of the digoxin antibody used in the kit. (orig.) [de

  18. Microfluidic Platform for Enzyme-Linked and Magnetic Particle-Based Immunoassay

    Directory of Open Access Journals (Sweden)

    Dorota G. Pijanowska

    2013-06-01

    Full Text Available This article presents design and testing of a microfluidic platform for immunoassay. The method is based on sandwiched ELISA, whereby the primary antibody is immobilized on nitrocelluose and, subsequently, magnetic beads are used as a label to detect the analyte. The chip takes approximately 2 h and 15 min to complete the assay. A Hall Effect sensor using 0.35-μm BioMEMS TSMC technology (Taiwan Semiconductor Manufacturing Company Bio-Micro-Electro-Mechanical Systems was fabricated to sense the magnetic field from the beads. Furthermore, florescence detection and absorbance measurements from the chip demonstrate successful immunoassay on the chip. In addition, investigation also covers the Hall Effect simulations, mechanical modeling of the bead–protein complex, testing of the microfluidic platform with magnetic beads averaging 10 nm, and measurements with an inductor-based system.

  19. Pregnancy does not affect HIV incidence test results obtained using the BED capture enzyme immunoassay or an antibody avidity assay.

    Directory of Open Access Journals (Sweden)

    Oliver Laeyendecker

    2010-10-01

    Full Text Available Accurate incidence estimates are needed for surveillance of the HIV epidemic. HIV surveillance occurs at maternal-child health clinics, but it is not known if pregnancy affects HIV incidence testing.We used the BED capture immunoassay (BED and an antibody avidity assay to test longitudinal samples from 51 HIV-infected Ugandan women infected with subtype A, C, D and intersubtype recombinant HIV who were enrolled in the HIVNET 012 trial (37 baseline samples collected near the time of delivery and 135 follow-up samples collected 3, 4 or 5 years later. Nineteen of 51 women were also pregnant at the time of one or more of the follow-up visits. The BED assay was performed according to the manufacturer's instructions. The avidity assay was performed using a Genetic Systems HIV-1/HIV-2 + O EIA using 0.1M diethylamine as the chaotropic agent.During the HIVNET 012 follow-up study, there was no difference in normalized optical density values (OD-n obtained with the BED assay or in the avidity test results (% when women were pregnant (n = 20 results compared to those obtained when women were not pregnant (n = 115; for BED: p = 0.9, generalized estimating equations model; for avidity: p = 0.7, Wilcoxon rank sum. In addition, BED and avidity results were almost exactly the same in longitudinal samples from the 18 women who were pregnant at only one study visit during the follow-up study (p = 0.6, paired t-test.These results from 51 Ugandan women suggest that any changes in the antibody response to HIV infection that occur during pregnancy are not sufficient to alter results obtained with the BED and avidity assays. Confirmation with larger studies and with other HIV subtypes is needed.

  20. Enzyme-immuno assay for total estrogens and human placental lactogen. Comparison with radio-immuno-assay in normal pregnancy-monitoring

    International Nuclear Information System (INIS)

    Raichvarg, D.; Tallet, F.; Lajeunie, E.; Bonnaire, Y.; Danglas, P.

    1980-01-01

    The concentrations of estrogens (E) and human placental lactogen (HLP) are estimated in sera by radio immuno-assay (RIA) and enzyme-immuno-assay (EIA). Statistical data indicate mean intra-assay variation coefficients of 7% and 12% for E and HLP tests, respectively. The correlation coefficient (RIA/EIA) are found higher than 0,9% for both hormonal assays. The dilution curves obtained by RIA and EIA are similar. However, Student'test gives a significant difference for E determination. In fact, total E and E 3 only are measured by EIA and RIA, respectively. In most cases biological interferences are negligible except for HLP in presence of higher protein or haemoglobin levels. RIA and EIA are performed to study serum HLP and E levels throughout normal pregnancies. Results allow to use EIA for HLP and E evaluations in pregnancy-monitoring [fr

  1. Sensitive electrochemical enzyme immunoassay microdevice based on architecture of dual ring electrodes with a sensing cavity chamber.

    Science.gov (United States)

    Dong, H; Li, C M; Zhou, Q; Sun, J B; Miao, J M

    2006-12-15

    A novel electrochemical detection architecture was investigated for enzyme immunoassay sensors. Microchips with dual-ring working and counter electrodes, and a sensing cavity chamber were made on glass slides. The glass surface of the microchip was coated by 3-aminopropyltriethoxysilane (APTES). Goat IgG, as a example, was covalently captured on APTES-modified glass surfaces through glutaraldehyde (GA) as a cross-linker. Enzyme substrate, p-aminophenyl phosphate (PAPP) was prepared by electrolysis. The enzyme conversion from home-synthetic PAPP to p-aminophenol (PAP) was examined by differential pulse voltammetry (DPV). A competitive inhibition enzyme-linked immunosorbant assay (ELISA) was designed to test the system. Experimental results demonstrate that a detection limit of 118 fg/ml of goat IgG and a dynamic range of 118 fg/ml to 1.18 ng/ml, up to five orders of magnitude could be achieved. Due to its novel architecture design and electronic detection scheme, the method can be used to fabricate portable electrochemical ELISA lab-on-chip systems. The technology could have great potential in clinical diagnostic applications.

  2. EIA publications directory, 1991

    International Nuclear Information System (INIS)

    1992-01-01

    Enacted in 1977, the Department of Energy (DOE) Organization Act established the Energy Information Administration (EIA) as the Department's independent statistical and analytical agency, with a mandate to collect and publish data and prepare analyses on energy production, consumption, prices, and resources, and projections of energy supply and demand. This edition of the EIA Publications Directory contains titles and abstracts of periodicals and one-time reports produced by the EIA from January through December 1991. This edition supplements EIA Publications Directory 1977--1989 and EIA Publications Directory 1990. The body of the Directory contains citations and abstracts arranged by broad subject categories, such as coal, petroleum, and natural gas and subcategories such as reserves, produces and byproducts, and marketing and economics. All reports are indexed alphabetically by subject and title and numerically by report number

  3. Screening for cocaine on Euro banknotes by a highly sensitive enzyme immunoassay.

    Science.gov (United States)

    Abdelshafi, Nahla A; Panne, Ulrich; Schneider, Rudolf J

    2017-04-01

    This study focused on quantitative detection of cocaine on Euro banknotes in Germany. A sensitive direct competitive immunoassay was developed and optimized with a limit of detection (LOD) of 5.6ng/L. Exhaustive cocaine extraction by solvent was tested using different methanol concentrations and buffered solutions. Cross-reactivity studies were performed to determine the degree of interference of cocaine metabolites with the immunoassay. Sixty-five Euro banknotes obtained from different districts in Berlin were evaluated. A 100% contamination frequency with cocaine was detected. A comparison between the amount of cocaine extracted by cotton swabbing of one square centimeter of the banknote showed a good correlation for lower contamination levels. This assay showed high sensitivity of detecting pg of cocaine per 1cm 2 of one banknote by swabbing 1cm 2 : 0, 14, and 21pg/cm 2 . Moreover, three notes of different denominations revealed high cocaine concentration; 1.1mg/note, and twice 55µg/note. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Novel enzyme immunoassay system for simultaneous detection of six subclasses of antiphospholipid antibodies for differential diagnosis of antiphospholipid syndrome.

    Science.gov (United States)

    Nojima, Junzo; Motoki, Yukari; Hara, Kazusa; Sakata, Toshiyuki; Ichihara, Kiyoshi

    2017-06-01

    : Antiphospholipid syndrome, which often complicates systemic lupus erythematosus (SLE), features high occurrence of arterial and/or venous thrombosis and recurrent fetal loss. However, which antibody subclass contributes to which clinical event remains uncertain. We newly developed an up-to-date enzyme immunoassay system using the AcuStar automated analyzer (Instrumentation Laboratory, Bedford, Massachusetts, USA) for parallel detection of six subclasses of antiphospholipid antibodies (aPLs): anticardiolipin antibodies (aCL) of IgG, IgM, and IgA and anti-β2-glycoprotein I antibodies (aβ2GPI) of IgG, IgM, and IgA. They were measured in 276 healthy volunteers and 138 patients with SLE: 45 with thromboembolic complications (29 arterial; 16 venous) and 93 without. Lupus anticoagulant activity in their plasma was measured according to the guidelines recommended by the Subcommittee on Lupus Anticoagulant/Phospholipid-Dependent Antibodies. aCL/β2GPI was measured with a standard ELISA kit commonly used in Japan. The positive results of IgG aCL, IgA aCL, and IgG aβ2GPI were closely associated with thromboembolic complications, whereas IgM aCL and IgM aβ2GPI were not. receiver operating characteristic analysis revealed that the accuracy of predicting thromboembolic complications based on the composite test results of the former three antibodies were obviously higher than by each alone. Regarding agreement with the test results of lupus anticoagulant activity, IgG aβ2GPI showed the closest match. Patients with SLE frequently possess various combinations of the six aPL subclasses, and this antibody spectrum is closely associated with thromboembolic events in these patients. This new automated enzyme immunoassay system allows simultaneous analysis of the profile of aPL subclasses for the differential diagnosis of antiphospholipid antibody syndrome in its early stage.

  5. Evaluation of the concomitant use of two different EIA tests for HIV screening in blood banks

    Directory of Open Access Journals (Sweden)

    Otani Marcia M.

    2003-01-01

    Full Text Available OBJECTIVE: In 1998, the Brazilian Ministry of Health made it mandatory for all blood banks in the country to screen donated blood for human immunodeficiency virus (HIV concomitantly using two different enzyme immunoassay (EIA tests. Concerned with the best use of available resources, our objective with this study was to evaluate the usefulness of conducting two EIA screening tests instead of just one. METHODS: We analyzed data from 1999 through 2001 obtained by testing 698 191 units of donated blood using two EIA HIV screening tests concomitantly at the Pro-Blood Foundation/Blood Center of São Paulo (Fundação Pró-Sangue/Hemocentro de São Paulo, which is a major blood center in the city of São Paulo, Brazil. All samples reactive in at least one of the two EIA tests were submitted for confirmation by a Western blot (WB test, and the persons who had donated those samples were also asked to return and provide a follow-up sample. RESULTS: Out of the 698 191 blood units that were donated, 2 718 of them (0.4% had to be discarded because they were reactive to at least one of the EIA tests. There were two WB-positive donation samples that were reactive in only one HIV EIA screening test. On their follow-up samples, both donors tested WB-negative. These cases were considered false positive results at screening. Of the 2 718 donors who were asked to return and provide a follow-up sample, 1 576 of them (58% did so. From these 1 576 persons, we found that there were two individuals who had been reactive to only one of the two EIA screening tests and who had also been negative on the WB at screening but who were fully seroconverted on the follow-up sample. We thus estimated that, in comparison to the use of a single EIA screening test, the use of two EIA screening tests would detect only one extra sample out of 410 700 units of blood. CONCLUSIONS: Our data do not support the use of two different, concomitant EIA screening tests for HIV. The great

  6. TRICLOSAN AND METHYL-TRICLOSAN MONITORING STUDY IN THE NORTHEAST OF SPAIN USING A MAGNETIC PARTICLE ENZYME IMMUNOASSAY AND CONFIRMATORY ANALYSIS BY GAS CHROMATOGRAPHY-MASS SPECTROMETRY

    Science.gov (United States)

    The occurrence of triclosan in the water environment around a Mediterranean region was investigated. Triclosan and methyl-triclosan content of ninety five environmental samples were screened using a magnetic particle enzyme immunoassay. Positive samples were confirmed by solid phase extraction (SPE...

  7. Immunoassay-Based Drug Tests Are Inadequately Sensitive for Medication Compliance Monitoring in Patients Treated for Chronic Pain.

    Science.gov (United States)

    Snyder, Marion L; Fantz, Corrine R; Melanson, Stacy

    2017-02-01

    Enzyme immunoassays (EIA) have notable limitations for monitoring therapeutic compliance in pain management. Chromatography coupled with mass spectrometry provides definitive results and superior sensitivity and specificity over traditional EIA testing. To analyze and compare the sensitivity of EIA results together with known prescriptions to liquid chromatography-tandem mass spectrometry (LC-MS/MS) for monitoring drug use (and abuse) in patients treated for chronic pain. A total of 530 urine samples from patients being treated for chronic pain were studied. Pain management clinic in the United States. The samples were tested for a profile of chronic pain medications and illicit drugs with commercially available EIA kits followed by analysis with Agilent LC-MS/MS system. The EIAs exhibited poor sensitivity and high rates of false negative results in the pain management setting. For example, 21% of EIA for opiates show false negative results. Mass spectrometry methods were more sensitive, detected a broader range of drugs and metabolites, and could detect non-prescribed drug use and simulations in compliance. Patients do not always accurately report drug use information, and some drugs do not have EIA methods available for comparative purposes. Mass spectrometry is a more robust and reliable method for detection of drugs used in the pain management setting. Due to the extent of undisclosed use and abuse of medications and illicit drugs, LC-MS/MS testing is necessary for adequate and accurate drug detection. In addition, LC-MS/MS methods are superior in terms of sensitivity and number of compounds that can be screened, making this a better method for use in pain management. Key words: Pain management, enzyme immunoassays, mass spectrometry, urine drug testing, prescription status, compliance.

  8. [Equine Infectious Anemia (EIA)].

    Science.gov (United States)

    Kaiser, A; Meier, H P; Straub, R; Gerber, V

    2009-04-01

    Equine Infectious Anemia (EIA) is a reportable, eradicable epizootic disease caused by the equine lentivirus of the retrovirus family which affects equids only and occurs worldwide. The virus is transmitted by blood, mainly by sanguivorous insects. The main symptoms of the disease are pyrexia, apathy, loss of body condition and weight, anemia, edema and petechia. However, infected horses can also be inapparent carriers without any overt signs. The disease is diagnosed by serological tests like the Coggins test and ELISA tests. Presently, Switzerland is offi cially free from EIA. However, Switzerland is permanently at risk of introducing the virus as cases of EIA have recently been reported in different European countries.

  9. EIA publications directory 1994

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1995-07-20

    Enacted in 1977, the Department of Energy (DOE) Organization Act established the Energy Information Administration (EIA) as the Department`s independent statistical and analytical agency, with a mandate to collect and publish data and prepare analyses on energy production, consumption, prices, resources, and projections of energy supply and demand. This edition of the EIA Publications Directory contains titles and abstracts of periodicals and one-time reports produced by EIA from January through December 1994. The body of the Directory contains citations and abstracts arranged by broad subject categories: metadata, coal, oil and gas, nuclear, electricity, renewable energy/alternative fuels, multifuel, end-use consumption, models, and forecasts.

  10. EIA publications directory, 1990

    International Nuclear Information System (INIS)

    1991-01-01

    Enacted in 1977, the Department of Energy (DOE) Organization Act established the Energy Information Administration (EIA) as the Department's independent statistical and analytical agency, with a mandate to collect and publish data and prepare analyses on energy production, consumption, prices, and resources, and projections of energy supply and demand. This edition of the EIA Publications Directory contains titles and abstracts of periodicals and one-time reports produced by the EIA from January through December 1990. This edition supplements EIA Publications Directory 1977--1989. The body of the Directory contains citations and abstracts arranged by broad subject categories, such as coal, petroleum, and natural gas and subcategories such as reserves, products and byproducts, and marketing and economics. All reports are indexed alphabetically by subject and title and numerically by report number

  11. EIA publications directory 1996

    International Nuclear Information System (INIS)

    1997-05-01

    This edition of the EIA Publications Directory contains titles and abstracts of periodicals and one-time reports produced by the Energy Information Administration (EIA) from January through December 1996. The body of the Directory contains citations and abstracts arranged by broad subject categories; metadata, coal, oil and gas, nuclear, electricity, renewable and energy/alternative fuels, multifuel, end-use consumption, models, and forecasts

  12. EIA publications directory 1996

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1997-05-01

    This edition of the EIA Publications Directory contains titles and abstracts of periodicals and one-time reports produced by the Energy Information Administration (EIA) from January through December 1996. The body of the Directory contains citations and abstracts arranged by broad subject categories; metadata, coal, oil and gas, nuclear, electricity, renewable and energy/alternative fuels, multifuel, end-use consumption, models, and forecasts.

  13. EIA Publications Directory 1995

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1995-10-01

    This directory contains abstracts and ordering information for individual issues of semiannual, annual, biennial, and triennial Energy Information Administration (EIA) periodicals, analysis reports, Service Reports, and model documentations. This edition covers periodicals and one-time reports produced by EIA from January through December 1995. The citations and abstracts are arranged by broad subject categories: metadata, coal, oil/gas, nuclear, electricity, renewable energy/alternative fuels, multifuel, end-use consumption, models, and forecasts. Indexes are provided.

  14. EIA publications directory 1997

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1998-04-01

    This edition of the EIA Publications Directory contains 68 titles and abstracts of periodicals and one time reports produced by EIA from January through December 1997. The body of the Directory contains citations and abstracts arranged by broad subject categories; (1) MetaData, (2) Coal, (3) Oil (4) Natural gas, (5) Nuclear, (6) Electricity, (7) Renewable energy and Alternative fuels, (8) Multifuel, (9) End use consumption, (10) Models, and (11) Forecasts.

  15. Enzyme-catalyzed silver deposition on irregular-shaped gold nanoparticles for electrochemical immunoassay of alpha-fetoprotein.

    Science.gov (United States)

    Lai, Wenqiang; Tang, Dianping; Que, Xiaohua; Zhuang, Junyang; Fu, Libing; Chen, Guonan

    2012-11-28

    A new and disposable electrochemical immunosensor was designed for detection of alpha-fetoprotein (AFP), as a model analyte, with sensitivity enhancement based on enzyme-catalyzed silver deposition onto irregular-shaped gold nanoparticles (ISGNPs). The assay was carried out with a sandwich-type immunoassay protocol by using ISGNP-labeled anti-AFP antibodies conjugated with alkaline phosphatase (ALP-Ab(2)) as detection antibodies. The enzymatically catalytic deposition of silver on the electrode could be measured by stripping analysis in KCl solution due to the Ag/AgCl solid-state voltammetric process. Several labeling protocols including spherical gold nanoparticle-labeled ALP-Ab(2) and ISGNP-labeled ALP-Ab(2) were investigated for determination of AFP, and improved analytical properties were achieved with the ISGNP labeling. With the ISGNP labeling method, the effects of incubation time and incubation temperature for antigen-antibody reaction, and deposition time of silver on the current responses of the electrochemical immunosensors were also monitored. Under optimal conditions, the electrochemical immunosensor exhibited a wide dynamic range from 0.01 ng mL(-1) to 200 ng mL(-1) with a detection limit of 5.0 pg mL(-1) AFP. The immunosensor displayed a good stability and acceptable reproducibility and accuracy. No significant differences at the 95% confidence level were encountered in the analysis of 10 clinical serum samples between the developed immunoassay and the commercially available electrochemiluminescent method for determination of AFP. Copyright © 2012 Elsevier B.V. All rights reserved.

  16. Peptide-Recombinant VP6 Protein Based Enzyme Immunoassay for the Detection of Group A Rotaviruses in Multiple Host Species.

    Directory of Open Access Journals (Sweden)

    Naveen Kumar

    Full Text Available We developed a novel enzyme immunoassay for the detection of group A rotavirus (RVA antigen in fecal samples of multiple host species. The assay is based on the detection of conserved VP6 protein using anti-recombinant VP6 antibodies as capture antibodies and anti-multiple antigenic peptide (identified and constructed from highly immunodominant epitopes within VP6 protein antibodies as detector antibodies. The clinical utility of the assay was evaluated using a panel of 914 diarrhoeic fecal samples from four different host species (bovine, porcine, poultry and human collected from diverse geographical locations in India. Using VP6- based reverse transcription-polymerase chain reaction (RT-PCR as the gold standard, we found that the diagnostic sensitivity (DSn and specificity (DSp of the new assay was high [bovine (DSn = 94.2% & DSp = 100%; porcine (DSn = 94.6% & DSp = 93.3%; poultry (DSn = 74.2% & DSp = 97.7% and human (DSn = 82.1% & DSp = 98.7%]. The concordance with RT-PCR was also high [weighted kappa (k = 0.831-0.956 at 95% CI = 0.711-1.0] as compared to RNA-polyacrylamide gel electrophoresis (RNA-PAGE. The performance characteristics of the new immunoassay were comparable to those of the two commercially available ELISA kits. Our results suggest that this peptide-recombinant protein based assay may serve as a preliminary assay for epidemiological surveillance of RVA antigen and for evaluation of vaccine effectiveness especially in low and middle income settings.

  17. Updates in immunoassays: parasitology.

    Science.gov (United States)

    Josko, Deborah

    2012-01-01

    Although most clinical laboratories use microscopy and routine O&P procedures when identifying parasitic infections, there are several parasites that are better detected through serological means. Toxoplasma, Giardia, and Cryptosporidium were discussed along with immunoassays used for their detection. Immunoassays provide quick results and are less labor intensive than specimen concentration and slide preparation for microscopic examination. These assays are easy to use and provide sensitive and specific results. Some clinical laboratories no longer perform O&Ps in house and refer specimens to reference laboratories for evaluation. By using immunoassays, some of the more common parasites can be identified in a timely manner reducing turn-around times. Some controversy exists over the use of IIF and EIA tests used for ANA testing along with measuring CRPs and PCT as predictors of bacterial sepsis and septic shock. Regardless of the methodology discussed in this series of articles, there are pros and cons to the various immunoassays available. Determining the most appropriate assay based on patient population and volume is governed by the institution and its patients' needs. In conclusion, immunoassays, whether manual or automated, are easy to use, cost effective and allow the medical laboratory professional to provide quick and accurate results to the clinician so the most appropriate treatment can be administered to the patient. The ultimate goal of healthcare professionals is to provide the highest quality of medical care in a timely manner. The use of immunoassays in the clinical laboratory allows the healthcare team to successfully achieve this goal.

  18. Anti-idiotypic nanobody-alkaline phosphatase fusion proteins: Development of a one-step competitive enzyme immunoassay for fumonisin B1 detection in cereal

    International Nuclear Information System (INIS)

    Shu, Mei; Xu, Yang; Liu, Xing; Li, Yanping; He, Qinghua; Tu, Zhui; Fu, Jinheng; Gee, Shirley J.; Hammock, Bruce D.

    2016-01-01

    A rapid and sensitive one-step competitive enzyme immunoassay for the detection of FB 1 was developed. The anti-idiotypic nanobody–alkaline phosphatase (Ab2β−Nb−AP) was validated by the AP enzyme activity and the properties of bounding to anti-FB1-mAb (3F11) through colorimetric and chemiluminescence analyses. The 50% inhibitory concentration and the detection limit (LOD) of colorimetric enzyme-linked immunosorbent assay (ELISA) for FB 1 were 2.69 and 0.35 ng mL −1 , respectively, with a linear range of 0.93–7.73 ng mL −1 . The LOD of the chemiluminescence ELISA (CLIA) was 0.12 ng mL −1 , and the IC 50 was 0.89 ± 0.09 ng mL −1 with a linear range of 0.29–2.68 ng mL −1 . Compared with LC-MS/MS, the results of this assay indicated the reliability of the Ab2β−Nb−AP fusion protein based one-step competitive immunoassay for monitoring FB 1 contamination in cereals. The Ab2β−Nb−AP fusion proteins have the potential to replace chemically-coupled probes in competitive enzyme immunoassay systems. - Highlights: • Ab2β−Nb−AP has the potential to replace chemically-coupled probes. • Ab2β−Nb−AP is homogeneous enzyme-labelled antigen can be prepared reproducibly. • We developed a green and rapid one-step competitive enzyme immunoassay. • The sensitivity of one-step CLIA was 9-folds higher than two-step ELISA.

  19. Anti-idiotypic nanobody-alkaline phosphatase fusion proteins: Development of a one-step competitive enzyme immunoassay for fumonisin B{sub 1} detection in cereal

    Energy Technology Data Exchange (ETDEWEB)

    Shu, Mei [State Key Laboratory of Food Science and Technology, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047 (China); Jiangxi-OAI Joint Research Institute, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047 (China); Xu, Yang, E-mail: xuyang@ncu.edu.cn [State Key Laboratory of Food Science and Technology, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047 (China); Jiangxi-OAI Joint Research Institute, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047 (China); Liu, Xing [State Key Laboratory of Food Science and Technology, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047 (China); College of Food Science and Technology, Hainan University, No. 58 Renmin Avenue, Haikou 570228 (China); Li, Yanping; He, Qinghua [Jiangxi-OAI Joint Research Institute, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047 (China); Tu, Zhui [State Key Laboratory of Food Science and Technology, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047 (China); Fu, Jinheng [Jiangxi-OAI Joint Research Institute, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047 (China); Gee, Shirley J.; Hammock, Bruce D. [Department of Entomology and UCD Comprehensive Cancer Center, University of California, Davis, CA 95616 (United States)

    2016-06-14

    A rapid and sensitive one-step competitive enzyme immunoassay for the detection of FB{sub 1} was developed. The anti-idiotypic nanobody–alkaline phosphatase (Ab2β−Nb−AP) was validated by the AP enzyme activity and the properties of bounding to anti-FB1-mAb (3F11) through colorimetric and chemiluminescence analyses. The 50% inhibitory concentration and the detection limit (LOD) of colorimetric enzyme-linked immunosorbent assay (ELISA) for FB{sub 1} were 2.69 and 0.35 ng mL{sup −1}, respectively, with a linear range of 0.93–7.73 ng mL{sup −1}. The LOD of the chemiluminescence ELISA (CLIA) was 0.12 ng mL{sup −1}, and the IC{sub 50} was 0.89 ± 0.09 ng mL{sup −1} with a linear range of 0.29–2.68 ng mL{sup −1}. Compared with LC-MS/MS, the results of this assay indicated the reliability of the Ab2β−Nb−AP fusion protein based one-step competitive immunoassay for monitoring FB{sub 1} contamination in cereals. The Ab2β−Nb−AP fusion proteins have the potential to replace chemically-coupled probes in competitive enzyme immunoassay systems. - Highlights: • Ab2β−Nb−AP has the potential to replace chemically-coupled probes. • Ab2β−Nb−AP is homogeneous enzyme-labelled antigen can be prepared reproducibly. • We developed a green and rapid one-step competitive enzyme immunoassay. • The sensitivity of one-step CLIA was 9-folds higher than two-step ELISA.

  20. Validation of an enzyme immunoassay for detection and semiquantification of cannabinoids in oral fluid.

    Science.gov (United States)

    Schwope, David M; Milman, Garry; Huestis, Marilyn A

    2010-06-01

    Oral fluid (OF) is gaining prominence as an alternative matrix for monitoring drugs of abuse in the workplace, criminal justice, and driving under the influence of drugs programs. It is important to characterize assay performance and limitations of screening techniques for Delta(9)-tetrahydrocannabinol (THC) in OF. We collected OF specimens by use of the Quantisal OF collection device from 13 daily cannabis users after controlled oral cannabinoid administration. All specimens were tested with the Immunalysis Sweat/OF THC Direct ELISA and confirmed by 2-dimensional GC-MS. The limit of detection was THC equivalent, and the assay demonstrated linearity from 1 to 50 microg/L, with semiquantification to 200 microg/L. Intraplate imprecision (n = 7) ranged from 2.9% to 7.7% CV, and interplate imprecision (n = 20) was 3.0%-9.1%. Cross-reactivities at 4 microg/L were as follows: 11-hydroxy-THC, 198%; Delta(8)-tetrahydrocannabinol (Delta(8)-THC), 128%; 11-nor-9-carboxy-THC (THCCOOH), 121%; THC (target), 98%; cannabinol, 87%; THCCOOH-glucuronide, 11%; THC-glucuronide, 10%; and cannabidiol, 2.4%. Of 499 tested OF specimens, 52 confirmed positive (THC 2.0-290 microg/L), with 100% diagnostic sensitivity at the proposed Substance Abuse and Mental Health Services Administration screening cutoff of 4 microg/L cannabinoids and GC-MS cutoff of 2 microg/L THC. Forty-seven specimens screened positive but were not confirmed by 2D-GC-MS, yielding 89.5% diagnostic specificity and 90.6% diagnostic efficiency. Thirty-one of 47 unconfirmed immunoassay positive specimens were from 1 individual and contained >400 ng/L THCCOOH, potentially contributing to cross-reactivity. The Immunalysis Sweat/OF THC Direct ELISA is an effective screening procedure for detecting cannabinoids in OF.

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

    NARCIS (Netherlands)

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

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

  2. Competitive chemiluminescent enzyme immunoassay for vitamin B12 analysis in human milk

    Science.gov (United States)

    BACKGROUND Few accurate data exist on the concentration of vitamin B12 in human milk. Binding of the vitamin to haptocorrin (HC) can interfere with the assay if not removed by pretreatment, and very low values can occur in women with poor B12 status. This study evaluated two competitive enzyme bind...

  3. Comparison of two enzyme immunoassays for the detection of Haemonchus contortus infections in sheep

    NARCIS (Netherlands)

    Schallig, H. D.; Hornok, S.; Cornelissen, J. B.

    1995-01-01

    Two enzyme-linked immunosorbent assays (ELISA) using either excretory/secretory (ES) products or crude somatic antigens (CSA) of adult Haemonchus contortus were compared for their ability to detect antibodies against H. contortus in sheep. Serum samples obtained from a group of 32 H. contortus

  4. Non-radiometric immunoassays fluoroimmunoassay (FIA) and fluorometric enzyme immunoassay (FEIA) and radioimmunoassay (RIA) for evaluation of thyroid function in normal and hypothyroid dogs

    Energy Technology Data Exchange (ETDEWEB)

    Jerico, M.M.; Larsson, C.E. [Sao Paulo Univ., SP (Brazil). Faculdade de Medicina Veterinaria e Zootecnia. Dept. de Clinica Medica]. E-mail: marciajerico@hotmail.com; Mendonca, B.B. [Sao Paulo Univ., SP (Brazil). Faculdade de Medicina . Lab. de Hormonios e Genetica Molecular; Otsuka, M. [Sao Paulo Univ., SP (Brazil). Faculdade de Medicina Veterinaria e Zootecnia. Hospital Veterinario; Maganin Junior, A. [Canil da Policia Militar do Estado de Sao Paulo, SP (Brazil)

    2001-07-01

    We proposed the comparison of thyroxine (T4) and free thyroxine (FT4) measurements by fluoroimmunoassay (FIA) and fluorometric enzyme immunoassay (FEIA) with radioimmunoassay (RIA) in thyroid function evaluation of normal (n=50) and hypothyroid dogs (n=9). T4 and FT4 serum concentrations were measured in basal conditions and 6 h after TRH stimulation (200 mug/IV). All our reference values are based on the 5th and 95th percentile. The reference values for basal T4 in healthy dogs were 0.50 to 2.35 mug/dL (FIA), 0.50 to 2.51 mug/dL (FEIA) and 0.35 to 0.74 mug/dL (RIA). After TRH, the values were >= 1.37 mug/dL (FIA), >= 0,26 mug/dL (FEIA) and >= 0.40 mug/dL (RIA). Basal FT4 values in healthy dogs were 0.65 to 2.20 ng/dL (FIA), 0.38 to 1.43 ng/dL (FEIA) and 0.10 to 1.24 ng/dL (RIA). After TRH, the values were >= 1.30 ng/dL (FIA), >= 0.77 ng/dL (FEIA) and >=0.50 ng/dL (RIA). In hypothyroid dogs, the mean +- SD for T4 in basal conditions and after TRH were 0.24 +- 0.20 mug/dL and 0.26 +- 0.20 mug/dL (FIA), 0.27 +- 0.12 mug/dL and 0.32 +- 0.51 mug/dL (FEIA) and 0.19 +- 0.30 mug/dL and 0,24 +- 0.09 mug/dL (RIA), respectively. In the same group the mean +- SD basal FT4 values and after TRH were 0.28 +- 0.33 ng/dL and 0.28 +- 0.39 ng/dL (FIA), 0,12 +- 0.26 ng/dL and 0.23 +- 0.56 ng/dL (FEIA) and 0,15 +- 0,15 ng/dL and 0,17 +- 0,28 ng/dL (RIA), respectively. Significant differences (p<0.05) between the normal and hypothyroid groups (Kruskall-Wallis test) were observed in the three methods and between basal and stimulated values in normal dogs (Wilcoxon test), by the three methods. The best sensitivity for diagnosing hypothyroidism was obtained through T4 values (100%), and the best specificity through FT4 values (100%), both determined by FIA after TRH stimulation. We conclude that T4 and FT4 measured by fluoroimmunoassay after TRH stimulation can be an excellent alternative. (author)

  5. Non-radiometric immunoassays (fluoroimmunoassay (FIA) and fluorometric enzyme immunoassay (FEIA) and radioimmunoassay (RIA) for evaluation of thyroid function in normal and hypothyroid dogs

    International Nuclear Information System (INIS)

    Jerico, M.M.; Larsson, C.E.

    2001-01-01

    We proposed the comparison of thyroxine (T4) and free thyroxine (FT4) measurements by fluoroimmunoassay (FIA) and fluorometric enzyme immunoassay (FEIA) with radioimmunoassay (RIA) in thyroid function evaluation of normal (n=50) and hypothyroid dogs (n=9). T4 and FT4 serum concentrations were measured in basal conditions and 6 h after TRH stimulation (200 mug/IV). All our reference values are based on the 5th and 95th percentile. The reference values for basal T4 in healthy dogs were 0.50 to 2.35 mug/dL (FIA), 0.50 to 2.51 mug/dL (FEIA) and 0.35 to 0.74 mug/dL (RIA). After TRH, the values were >= 1.37 mug/dL (FIA), >= 0,26 mug/dL (FEIA) and >= 0.40 mug/dL (RIA). Basal FT4 values in healthy dogs were 0.65 to 2.20 ng/dL (FIA), 0.38 to 1.43 ng/dL (FEIA) and 0.10 to 1.24 ng/dL (RIA). After TRH, the values were >= 1.30 ng/dL (FIA), >= 0.77 ng/dL (FEIA) and >=0.50 ng/dL (RIA). In hypothyroid dogs, the mean +- SD for T4 in basal conditions and after TRH were 0.24 +- 0.20 mug/dL and 0.26 +- 0.20 mug/dL (FIA), 0.27 +- 0.12 mug/dL and 0.32 +- 0.51 mug/dL (FEIA) and 0.19 +- 0.30 mug/dL and 0,24 +- 0.09 mug/dL (RIA), respectively. In the same group the mean +- SD basal FT4 values and after TRH were 0.28 +- 0.33 ng/dL and 0.28 +- 0.39 ng/dL (FIA), 0,12 +- 0.26 ng/dL and 0.23 +- 0.56 ng/dL (FEIA) and 0,15 +- 0,15 ng/dL and 0,17 +- 0,28 ng/dL (RIA), respectively. Significant differences (p<0.05) between the normal and hypothyroid groups (Kruskall-Wallis test) were observed in the three methods and between basal and stimulated values in normal dogs (Wilcoxon test), by the three methods. The best sensitivity for diagnosing hypothyroidism was obtained through T4 values (100%), and the best specificity through FT4 values (100%), both determined by FIA after TRH stimulation. We conclude that T4 and FT4 measured by fluoroimmunoassay after TRH stimulation can be an excellent alternative. (author)

  6. Evaluation of nine immunoassay kits (enzyme immunoassay and direct fluorescence) for detection of Giardia lamblia and Cryptosporidium parvum in human fecal specimens.

    OpenAIRE

    Garcia, L S; Shimizu, R Y

    1997-01-01

    It is well known that Giardia lamblia and Cryptosporidium parvum can cause severe symptoms in humans, particularly those who are immunologically compromised. Immunoassay procedures offer both increased sensitivity and specificity compared to conventional staining methods. These reagents are also helpful when screening large numbers of patients, particularly in an outbreak situation or when screening patients with minimal symptoms. The data obtained by using 9 diagnostic kits were compared: di...

  7. Automated thermometric enzyme immunoassay of human proinsulin produced by Escherichia coli

    International Nuclear Information System (INIS)

    Birnbaum, S.; Buelow, L.; Hardy, K.; Danielsson, B.; Mosbach, K.

    1986-01-01

    The authors have determined and monitored the production and release of human proinsulin by genetically engineered Escherichia coli cells. Several M9 media samples were analyzed sequentially after centrifugation with the aid of a rapid automated flow-through thermometric enzyme-linked immunosorbent assay (TELISA) system. The response time was 7 min after after sample injection and a single assay was complete after 13 min. Insulin concentrations in the range of 0.1-50 μg/ml could be determined. The TELISA method correlated well with conventional radioimmunoassay determinations. Standard curves were reproducible over a period of several days even when the immobilized antibody column was stored at 25 0 C in the enzyme thermistor unit. Thus, immediate assay start up was possible

  8. Ultrasensitive electrochemical immunoassay of staphylococcal enterotoxin B in food using enzyme-nanosilica-doped carbon nanotubes for signal amplification.

    Science.gov (United States)

    Tang, Dianping; Tang, Juan; Su, Biling; Chen, Guonan

    2010-10-27

    A new sandwich-type electrochemical immunoassay for ultrasensitive detection of staphylococcal enterotoxin B (SEB) in food was developed using horseradish peroxidase-nanosilica-doped multiwalled carbon nanotubes (HRPSiCNTs) for signal amplification. Rabbit polyclonal anti-SEB antibodies immobilized on the screen-printed carbon electrode (SPCE) and covalently bound to the HRPSiCNTs were used as capture antibodies and detection antibodies, respectively. In the presence of SEB analyte, the sandwich-type immunocomplex could be formed between the immobilized anti-SEB on the SPCE and anti-SEB-labeled HRPSiCNTs, and the carried HRP could catalyze the electrochemical reduction of H2O2 with the help of thionine. The high content of HRP in the HRPSiCNTs could greatly amplify the electrochemical signal. Under optimal conditions, the reduction current increased with the increase of SEB in the sample, and exhibited a dynamic range of 0.05-15 ng/mL with a low detection limit (LOD) of 10 pg/mL SEB (at 3σ). Intra- and interassay coefficients of variation were below 10%. In addition, the assay was evaluated with SEB spiked samples including watermelon juice, soymilk, apple juice, and pork food, receiving excellent correlation with results from commercially available enzyme-linked immunosorbent assay (ELISA).

  9. [Fundamental evaluation of apolipoprotein B-48 by chemiluminescence enzyme immunoassay--identification of apolipoprotein B-48 with immunoblotting].

    Science.gov (United States)

    Sato, Itsuko; Fujioka, Yoshio; Hayashi, Fujio; Mukai, Masahiko; Kawano, Seiji; Ishikawa, Yuichi; Yamashita, Shizuya; Kumagai, Shunichi

    2007-06-01

    Apolipoprotein B-48 (apo B-48) is a constituent of chylomicrons and chylomicron remnants, and its fasting concentration has been reported to be a marker of postprandial hyperlipidemia, which is thought to be a risk factor of atherosclerosis. We evaluated the serum apo B-48 concentrations by chemiluminescence enzyme immunoassay (CLEIA), which was recently introduced as Lumipulse f fully automated immunosaasy analyzer by Fujirebio Inc (Tokyo, Japan), and performed immunoblotting on agarose gel electrophoresis with anti-apo B-48 antibody. Apo B-48 assay was intra-assay reproducible (CVs: 1.9-3.1%) and inter-assay reproducible (CVs: 2.2-4.4%). The assay range for apo B-48 was from 0.2 to 40.0 microg/ml. The effects of interfering substances such as free/conjugated birirubin, hemoglobin, Intrafat, ascorbic acid and rheumatoid factor were negligible. For storage, it was preferable to freeze, and to avoid frozen-thaw process as much as possible. Anti-apo B-48 antibody was reactive over a wide range from origin to the position of very-low-density lipoproteins in immunoblotting after agarose gel electrophoresis. Apo B-48 measurement by CLEIA was feasible to clinical use for the assessment of lipoprotein metabolism.

  10. Development and validation of an indirect enzyme immunoassay for detection of antibody to Brucella abortus in milk.

    Science.gov (United States)

    Nielsen, K; Smith, P; Gall, D; Perez, B; Cosma, C; Mueller, P; Trottier, J; Cote, G; Boag, L; Bosse, J

    1996-09-01

    An indirect enzyme immunoassay for detection of antibody to Brucella abortus in bovine milk was developed and validated using 6238 milk samples from Canadian herds (brucellosis free) and 202 samples from herds infected with B. abortus (from Argentina and Chile). The assay utilized lipopolysaccharide as the antigen, immobilized on the polystyrene matrix, whole milk to test and a mouse monoclonal antibody, specific for an epitope of bovine IgG1, conjugated with horseradish peroxidase. The sensitivity of the assay was 95.2% +/- 3.7% at a confidence limit of 95% for samples from B. abortus infected herds obtained from chile and 98.7% +/- 0.3% at a confidence limit of 95% for samples from similar herds in Argentina. Of the negative milk samples tested, 77 gave a result above the threshold value of 0.200 optical density units. When the 77 false positive samples were retested using 7.5 mM (final concentration) of EDTA and ethyleneglycol-bis-aminoether-N,N,N', N'-tetraacetic acid (EGTA), the number of false positive reactions was reduced to 3, giving a diagnostic specificity of 99.95%. The divalent cation chelating agents did not affect positive reactions and the sensitivity remained the same. Based on control samples included with each assay, the performance of the assay was consistent.

  11. Summary of field trials using the direct and competitive enzyme immunoassays for detection of antibody to brucella abortus

    International Nuclear Information System (INIS)

    Nielsen, K.; Gall, D.

    1998-01-01

    Two indirect and two competitive enzyme immunoassays for detection of antibody to Brucella abortus, validated elsewhere, were field tested in five different Latin American laboratories. Testing was performed according to standardised protocols using sera obtained in each area. Sera from B. abortus infected herds, from vaccinated (but serologically negative in a screening test) and non-vaccinated cattle were tested in each assay and compared to the results obtained with conventional diagnostic tests used for diagnosis of brucellosis in each country. Relative sensitivity and specificity values were calculated for each country as well as a weighted summary combining the data from all the participating laboratories. The result demonstrate that all ELISAs performed as well as, or better than, the conventional aerological tests. Given the inherent errors in the use of the latter in the diagnosis of brucellosis, it is recommended that the ELISAs described here be considered as replacements for the conventional tests. The CELISA using the lipopolysaccharide antigen with the competing monoclonal antibody M84, should be considered as the most useful because of cross-species and vaccination considerations. (author)

  12. Development of enzyme immunoassays (ELISA and Western blot) for the serological diagnosis of dermatophytosis in symptomatic and asymptomatic cats.

    Science.gov (United States)

    Santana, Aline Elisa; Taborda, Carlos Pelleschi; Severo, Julia So; Rittner, Glauce Mary Gomes; Muñoz, Julian Esteban; Larsson, Carlos Eduardo; Larsson, Carlos Eduardo

    2018-01-01

    Dermatophytosis is the most common fungal infection in cats worldwide and plays an important role in both animal and human health due to their high zoonotic potential. Effective screening is a strong preventive measure and the fungal culture is quite useful but requires full laboratorial experience and it takes a long time to obtain the result. A rapid and accurate screening test for dermatophytosis in cats is crucial for the effective control of disease outbreaks. The aim of this study was to develop and evaluate the diagnostic efficacy of enzyme immunoassays (ELISA and Western blot [WB]) for the rapid and precise diagnosis of dermatophytosis in cats. Seventy cats of various ages were divided into three groups: S (symptomatic, n = 20), AS (asymptomatic, n = 30), and N (negative, n = 20). All animals were submitted to fungal culture and blood samples for carrying out the serological tests. A significant difference (P technique detected 13 bands, and the 50 kDa protein was considered the most immunogenic protein, observing reactivity in 83.3% in the symptomatic group and 66.6% in the asymptomatic group. The study concluded that ELISA and WB were useful tools to reliably detect cats that have been exposed to M. canis. © The Author 2017. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  13. Validation of enzyme immunoassay applied to the serological diagnosis of hog brucellosis

    International Nuclear Information System (INIS)

    Ramondino, Romina; Marticorena, Damian

    1997-01-01

    The application of an enzyme immunization test in the serological diagnostic of a disease demands its validation as a previous stage. A test validation requires to know: cut off, diagnostic and relative specificity and sensitivity and harmonization grade between the new test and the classical test in use. The purpose of this work is the following: a) To determine the cut off and to calculate the specificity of two immunization tests, indirect (I-ELISA) and competitive (C-ELISA) by the analysis of serum coming from Canada (country free of this disease); b) To calculate the values of relative sensitivity of the tests with serum coming from infected populations and diagnostic sensitivity with samples of serum from hogs with positive isolation to Brucella suis

  14. Rapid and specific enzyme immunoassay on hydrophobic grid membrane filter for detection and enumeration of thermophilic Campylobacter spp. from milk and chicken rinses.

    Science.gov (United States)

    Wang, H; Boyle, E; Farber, J

    2000-04-01

    Six commercially available anti-Campylobacter antibodies were examined for their applicability in an enzyme immunoassay on hydrophobic grid membrane filters, both for the detection and enumeration of Campylobacter spp. When a panel of nine Campylobacter (seven Campylobacter jejuni and two Campylobacter coli) and eight non-Campylobacter strains were used in a dot-blot format enzyme immunoassay to test the specificity of these antibodies, only one polyclonal antibody (Biodesign) detected all Campylobacter strains. Escherichia coli O157:H7 produced weak nonspecific signals due to endogenous peroxidase activity. The specificity of this Biodesign antibody was further tested against 30 more Campylobacter strains and more than 600 non-Campylobacter strains on hydrophobic grid membrane filters grown on modified Campylobacter agar with charcoal and deoxycholate, a Campylobacter selective medium. All the Campylobacter strains were detected, whereas only two (Acinetobacter calcoaceticus, Salmonella Minnesota) of the approximately 130 non-Campylobacter strains, which grew on modified Campylobacter agar with charcoal and deoxycholate, gave false-positive signals. This simple, rapid, and specific enzyme immunoassay also detected Campylobacter spp. from inoculated milk and chicken rinses and naturally contaminated chicken rinses.

  15. EIA Publications Directory 1993

    Energy Technology Data Exchange (ETDEWEB)

    1994-07-18

    This directory contains abstracts and ordering information for EIA publications released in the above time period. The abstracts are arranged by broad subject category such as coal, petroleum, natural gas, and electric power. A comprehensive subject index, a title index, and a report number index are included. Each entry gives the title, report number, publication frequency, date, number of pages, and ordering information.

  16. EIA publications directory, 1992

    International Nuclear Information System (INIS)

    1993-01-01

    This directory contains abstracts and ordering information for EIA publications. The abstracts are arranged by broad subject category such as coal, petroleum, natural gas, and electric power. A comprehensive subject index, a title index, and a report number index are included. Each entry gives the title, report number, publication frequency, date, number of pages, and ordering information. Publication began with the 1979 edition

  17. EIA Publications Directory 1993

    International Nuclear Information System (INIS)

    1994-01-01

    This directory contains abstracts and ordering information for EIA publications released in the above time period. The abstracts are arranged by broad subject category such as coal, petroleum, natural gas, and electric power. A comprehensive subject index, a title index, and a report number index are included. Each entry gives the title, report number, publication frequency, date, number of pages, and ordering information

  18. Development and application of an immunoaffinity column enzyme immunoassay for mycotoxin zearalenone in complicated samples.

    Directory of Open Access Journals (Sweden)

    Xiaoqian Tang

    Full Text Available The zearalenone (ZEA monoclonal antibody (mAb 2D3, one of the highest sensitivity antibodies, was developed. Based on this mAb, it was established of an immunoaffinity column (IAC coupled with an indirect competitive enzyme-linked immunosorbent assay (icELISA. After optimization, the icELISA allowed an IC50 against ZEA of 0.02 µg L(-1. The mAb 2D3 exhibited a high recognition of ZEA (100% and β-zearalenol (β-ZOL, 88.2%. Its cross-reactivity with α-zearalenol (α-ZOL and β-zearalanol (β-ZAL were found to be 4.4% and 4.6%, respectively. The IAC-icELISA method was employed to analyze ZEA contamination in food samples, compared with high-performance liquid chromatography (HPLC. The spiked assay for ZEA demonstrated the considerable recoveries for IAC-icELISA (83-93% and HPLC (94-108% methods. Results showed that the mAb 2D3 and IAC-icELISA method posed potential applications in sensitively determination of ZEA in maize.

  19. Development and application of an immunoaffinity column enzyme immunoassay for mycotoxin zearalenone in complicated samples.

    Science.gov (United States)

    Tang, Xiaoqian; Li, Xin; Li, Peiwu; Zhang, Qi; Li, Ran; Zhang, Wen; Ding, Xiaoxia; Lei, Jiawen; Zhang, Zhaowei

    2014-01-01

    The zearalenone (ZEA) monoclonal antibody (mAb) 2D3, one of the highest sensitivity antibodies, was developed. Based on this mAb, it was established of an immunoaffinity column (IAC) coupled with an indirect competitive enzyme-linked immunosorbent assay (icELISA). After optimization, the icELISA allowed an IC50 against ZEA of 0.02 µg L(-1). The mAb 2D3 exhibited a high recognition of ZEA (100%) and β-zearalenol (β-ZOL, 88.2%). Its cross-reactivity with α-zearalenol (α-ZOL) and β-zearalanol (β-ZAL) were found to be 4.4% and 4.6%, respectively. The IAC-icELISA method was employed to analyze ZEA contamination in food samples, compared with high-performance liquid chromatography (HPLC). The spiked assay for ZEA demonstrated the considerable recoveries for IAC-icELISA (83-93%) and HPLC (94-108%) methods. Results showed that the mAb 2D3 and IAC-icELISA method posed potential applications in sensitively determination of ZEA in maize.

  20. Determination of residual ractopamine concentrations by enzyme immunoassay in treated pig's tissues on days after withdrawal.

    Science.gov (United States)

    Pleadin, Jelka; Perši, Nina; Vulić, Ana; Milić, Dinka; Vahčić, Nada

    2012-03-01

    The objective of this study was to measure residual ractopamine concentrations in tissues of pigs as experimental animals after treatment with dietary ractopamine for 28 consecutive days. Ractopamine was administered orally to the experimental group (n=9) in a dose of 0.1mg/kg body mass per day, whereas control animals (n=3) were left untreated. Treated pigs (60kg) were sacrificed on days 1, 3 and 8 of treatment discontinuation and residues were determined in kidney, liver, muscle, brain and heart tissues using previously validated enzyme-linked immunosorbent assay (ELISA) as a quantitative screening method. Validation showed good mean recoveries (approx. 70-90%) with acceptable inter- and intra-day relative standard deviations (RSDractopamine tissue concentrations. The highest ractopamine concentration on day 1 (24h) after the last exposure was recorded in the kidney (12.49±7.96ng/g), followed by the liver (7.21±2.73ng/g), heart (1.26±0.12ng/g) and brain (0.63±0.05ng/g); at this time of withdrawal, residues were not detected in the muscle. Ractopamine was depleted rapidly from all study tissues, with mostly no detectable residues on day 8 of withdrawal. Copyright © 2011 Elsevier Ltd. All rights reserved.

  1. Development of a sensitive enzyme immunoassay (ELISA for specific identification of Lachesis acrochorda venom

    Directory of Open Access Journals (Sweden)

    V Núñez Rangel

    2012-01-01

    Full Text Available The snake genus Lachesis provokes 2 to 3% of snakebites in Colombia every year. Two Lachesis species, L. acrochorda and L. muta, share habitats with snakes from another genus, namely Bothrops asper and B. atrox. Lachesis venom causes systemic and local effects such as swelling, hemorrhaging, myonecrosis, hemostatic disorders and nephrotoxic symptoms similar to those induced by Bothrops, Portidium and Bothriechis bites. Bothrops antivenoms neutralize a variety of Lachesis venom toxins. However, these products are unable to avoid coagulation problems provoked by Lachesis snakebites. Thus, it is important to ascertain whether the envenomation was caused by a Bothrops or Lachesis snake. The present study found enzyme linked immunosorbent assay (ELISA efficient for detecting Lachesis acrochorda venom in a concentration range of 3.9 to 1000 ng/mL, which did not show a cross-reaction with Bothrops, Portidium, Botriechis and Crotalus venoms. Furthermore, one fraction of L. acrochorda venom that did not show crossreactivity with B. asper venom was isolated using the same ELISA antibodies; some of its proteins were identified including one Gal-specific lectin and one metalloproteinase. This test may be useful to physicians, since it could be applicable for tracking the kinetic distribution of antigens in patients or experimentally envenomed animals.

  2. Enzyme Immunoassays for Serological Diagnosis of Bovine Brucellosis: A Trial in Latin America

    Science.gov (United States)

    Gall, D.; Colling, A.; Marino, O.; Moreno, E.; Nielsen, K.; Perez, B.; Samartino, L.

    1998-01-01

    The results of a field trial conducted in Latin America with two indirect enzyme-linked immunosorbent assays (ELISAs) and two competitive ELISAs (CELISAs) for the detection of bovine antibody to Brucella abortus are reported. One of the CELISA formats performed most accurately. The percentage of positive reactions in the CELISA relative to the selected positive rose bengal agglutination test (RBT) and complement fixation test (CFT) results was 97.47%, the percentage of negatives relative to the selected negative RBT and CFT results for unexposed cattle was 98.32%, and the percentage of negatives in cattle vaccinated with B. abortus 19 was 96.51%. The same assay format under Canadian conditions had an actual sensitivity of 100%, a specificity of 99.90% in nonvaccinates, and a specificity of 97.7% in a strain 19-vaccinated population. Overall, the CELISA performed as expected and the results were not dissimilar from the results obtained in the Canadian study. This provided further evidence that this CELISA can in many instances differentiate infected cattle from those that are vaccinated or infected with a cross-reacting organism while still giving very few false-positive or false-negative results. PMID:9729532

  3. Evaluation of a malaria antibody enzyme immunoassay for use in blood screening

    Directory of Open Access Journals (Sweden)

    Jun Seo Oh

    2008-02-01

    Full Text Available Transfusion-transmitted malaria is rare, but it may produce severe problem in the safety of blood transfusion due to the lack of reliable procedure to evaluate donors potentially exposed to malaria. Here, we evaluated a new enzyme-linked immunosorbent assay malaria antibody test (ELISA malaria antibody test, DiaMed, Switzerland to detect antibodies to Plasmodium vivax (the indigenous malaria in the blood samples in the Republic of Korea (ROK. Blood samples of four groups were obtained and analyzed; 100 samples from P.vivax infected patients, 35 from recovery patients, 366 from normal healthy individuals, and 325 from domestic travelers of non-endemic areas residents to risky areas of ROK. P.vivax antibody levels by ELISA were then compared to the results from microscopic examination and polymerase chain reaction (PCR test. As a result, the ELISA malaria antibody test had a clinical sensitivity of 53.0% and a clinical specificity of 94.0% for P.vivax. Twenty out of 325 domestic travelers (6.2% were reactive and 28 cases (8.6% were doubtful. Of the reactive and doubtful cases, only two were confirmed as acute malaria by both microscopy and PCR test. Thus we found that the ELISA malaria antibody test was insufficiently sensitive for blood screening of P.vivax in ROK.

  4. Chronic Chagas Disease Diagnosis: A Comparative Performance of Commercial Enzyme Immunoassay Tests

    Science.gov (United States)

    Santos, Fred Luciano Neves; de Souza, Wayner Vieira; da Silva Barros, Michelle; Nakazawa, Mineo; Krieger, Marco Aurélio; de Miranda Gomes, Yara

    2016-01-01

    There is a significant heterogeneity in reported performance of serological assays for Chagas disease diagnosis. The conventional serology testing in laboratory diagnosis and in blood banks is unsatisfactory because of a high number of inconclusive and misclassified results. We aimed to assess the quality of four commercially available enzyme-linked immunosorbent assay tests for their ability to detect Trypanosoma cruzi antibodies in 685 sera samples. Cross-reactivity was assessed by using 748 sera from patients with unrelated diseases. Initially, we found that the reactivity index against T. cruzi antigen was statistically higher in sera from Chagas disease patients compared with those from non-chagasic patients, supporting the notion that all evaluated tests have a good discriminatory ability toward the diagnosis of T. cruzi infection in patients in the chronic phase of the disease. Although all tests were similarly sensitive for diagnosing T. cruzi infection, there were significant variations in terms of specificity and cross-reactivity among them. Indeed, we obtained divergent results when testing sera from patient with unrelated diseases, particularly leishmaniasis, with the levels of cross-reactivity being higher in tests using whole T. cruzi extracts compared with those using recombinant proteins. Our data suggest that all four tests may be used for the laboratory diagnosis and routine blood screening diagnose for Chagas disease. We also emphasize that, despite their general good performance, caution is needed when analyzing the results when these tests are performed in areas where other diseases, particularly leishmaniasis, are endemic. PMID:26976886

  5. Evaluation of the highly sensitive chemiluminescent enzyme immunoassay "Lumipulse HBsAg-HQ" for hepatitis B virus screening.

    Science.gov (United States)

    Deguchi, Matsuo; Kagita, Masanori; Yoshioka, Nori; Tsukamoto, Hiroko; Takao, Miyuki; Tahara, Kazuko; Maeda, Ikuhiro; Hidaka, Yoh; Yamauchi, Satoshi; Kaneko, Atsushi; Miyakoshi, Hideo; Isomura, Mitsuo

    2017-10-06

    Ongoing efforts in the development of HBsAg detection kits are focused on improving sensitivity and specificity. The purpose of this study was to evaluate an improved, highly sensitive quantitative assay, "Lumipulse HBsAg-HQ", a chemiluminescent enzyme immunoassay designed for a fully automated instrument, the "Lumipulse G1200". Serum samples for reproducibility, dilution, correlation, sensitivity, and specificity studies were obtained from patients at the Osaka University Hospital. Seroconversion and sensitivity panels were purchased from a commercial vender. Subtype, sensitivity panels, and HBsAg recombinant proteins with one or two amino acid substitutions were prepared in-house. The coefficients of variation for the low, medium, and high concentration samples ranged from 1.93 to 2.55%. The HBsAg-HQ reagent for dilution testing showed good linearity in the 0.005-150 HBsAg IU/mL range and no prozone phenomenon. All 102 HBV carrier samples were positive by HBsAg-HQ, while other commercial reagents showed one or more to be negative. In the seroconversion panel, the 14-day blood sample was positive. The sensitivity against HBsAg-HQ "ad" and "ay" subtypes was 0.025 ng/mL. Comparisons among the HBsAg-HQ, HISCL, and Architect HBsAg reagents were performed using the Bland-Altman plot. Specificity for 1000 seronegative individuals was 99.7%. HBsAg-HQ detected 29 positive serum among 12 231 routinely obtained serum samples, which showed concentrations of 0.005-0.05 HBsAg IU/mL. According to these results, the Lumipulse HBsAg-HQ assay, with a highly sensitive limit of detection of 0.005 IU/mL, may facilitate the development of a better management strategy for a considerable proportion of infected patients. © 2017 Wiley Periodicals, Inc.

  6. Anti-cyclic citrullinated peptide autoantibodies measured by an automated enzyme immunoassay: analytical performance and clinical correlations.

    Science.gov (United States)

    Tampoia, Marilina; Brescia, Vincenzo; Fontana, Antonietta; Maggiolini, Piera; Lapadula, Giovanni; Pansini, Nicola

    2005-05-01

    Autoantibodies against cyclic citrullinated peptide (anti-CCP) are considered to be a sensitive and specific marker for rheumatoid arthritis (RA). This study evaluated the analytical performance and clinical correlation of an automated enzyme immunoassay (DSX, DINEX Technologies), for the detection of anti-CCP autoantibodies (DIASTAT anti-CCP, Axis-Shield, DUNDEE UK). Commercial controls and serum pools were used to determine its precision, analytical sensitivity, functional sensitivity and linearity. Sera from 83 patients with established RA and from 140 controls, including patients with various autoimmune diseases, viral infections and cancer, as well as sex- and age-matched healthy subjects, were studied. The rheumatoid factor (RF) was also assayed in each sample, and the results were compared to the anti-CCP findings. The total imprecision (CV%) was 4.7-7.2% for concentrations ranging between 1.98 and 71.81 U/mL. The lower detection limit was 0.038 U/mL. At a cut-off of 5 U/mL, the sensitivity and specificity for RA were 67.5% and 99.3%, respectively. The RF had a sensitivity of 66.3% and a lower specificity 82.1% than anti-CCP. When the two antibodies were used together, the specificity was 99.1%. The anti-CCP assay we examined on a fully automated system showed a good analytical performance (analytical and functional sensitivity, linearity) and good clinical correlation. We conclude that this system can provide rapid, useful data.

  7. Development of improved enzyme-based and lateral flow immunoassays for rapid and accurate serodiagnosis of canine brucellosis.

    Science.gov (United States)

    Cortina, María E; Novak, Analía; Melli, Luciano J; Elena, Sebastián; Corbera, Natalia; Romero, Juan E; Nicola, Ana M; Ugalde, Juan E; Comerci, Diego J; Ciocchini, Andrés E

    2017-09-01

    Brucellosis is a widespread zoonotic disease caused by Brucella spp. Brucella canis is the etiological agent of canine brucellosis, a disease that can lead to sterility in bitches and dogs causing important economic losses in breeding kennels. Early and accurate diagnosis of canine brucellosis is central to control the disease and lower the risk of transmission to humans. Here, we develop and validate enzyme and lateral flow immunoassays for improved serodiagnosis of canine brucellosis using as antigen the B. canis rough lipopolysaccharide (rLPS). The method used to obtain the rLPS allowed us to produce more homogeneous batches of the antigen that facilitated the standardization of the assays. To validate the assays, 284 serum samples obtained from naturally infected dogs and healthy animals were analyzed. For the B. canis-iELISA and B. canis-LFIA the diagnostic sensitivity was of 98.6%, and the specificity 99.5% and 100%, respectively. We propose the implementation of the B. canis-LFIA as a screening test in combination with the highly accurate laboratory g-iELISA. The B. canis-LFIA is a rapid, accurate and easy to use test, characteristics that make it ideal for the serological surveillance of canine brucellosis in the field or veterinary laboratories. Finally, a blind study including 1040 serum samples obtained from urban dogs showed a prevalence higher than 5% highlighting the need of new diagnostic tools for a more effective control of the disease in dogs and therefore to reduce the risk of transmission of this zoonotic pathogen to humans. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Radiolabelling for immunoassay

    International Nuclear Information System (INIS)

    Chapman, R.S.

    1998-01-01

    Since the early 1960s labelled compounds employed in immunoassay techniques, both radioimmunoassay and immunoradiometric assay, have involved radioisotopes typically 3 H (tritium) and 125 Iodine. With the advent of increasingly stringent governmental regulations regarding usage and disposal of radioisotopes and the impetus of research towards improved immunoassay sensitivity following the discovery of monoclonal antibodies and their application to excess reagent immunometric assay methodology, radioisotopic labels are gradually being replaced by non-isotopic labels: enzyme, fluorescence and chemiluminescence

  9. Application of a Newly Developed High-Sensitivity HBsAg Chemiluminescent Enzyme Immunoassay for Hepatitis B Patients with HBsAg Seroclearance

    OpenAIRE

    Shinkai, Noboru; Matsuura, Kentaro; Sugauchi, Fuminaka; Watanabe, Tsunamasa; Murakami, Shuko; Iio, Etsuko; Ogawa, Shintaro; Nojiri, Shunsuke; Joh, Takashi; Tanaka, Yasuhito

    2013-01-01

    We modified and automated a highly sensitive chemiluminescent enzyme immunoassay (CLEIA) for surface antigen (HBsAg) detection using a combination of monoclonal antibodies, each for a specific epitope of HBsAg, and by improving an earlier conjugation technique. Of 471 hepatitis B virus (HBV) carriers seen in our hospital between 2009 and 2012, 26 were HBsAg seronegative as determined by the Abbott Architect assay. The Lumipulse HBsAg-HQ assay was used to recheck those 26 patients who demonstr...

  10. [DIAGNOSTIC VALUE OF COMBINED USE OF COMBINED METHOD OF ENZYME IMMUNOASSAY AND POLYMERASE CHAIN REACTION TO DETECT OF INTRAUTERINE FETAL INFECTION BY PARVOVIRUS B19].

    Science.gov (United States)

    Bondarenko, N P; Lakatosh, V P; Lakatosh, P V; Malanchuk, O B; Poladich, I V

    2015-01-01

    The combined method of diagnosis parvovirus infection during pregnancy by maternal serum enzyme immunoassay and deoxyribonucleic acid isolation parvovirus B19 polymerase chain reaction in amnniotic fluid and fetal cord blood newborns, can diagnose vertical transmission and anticipate a negative effect on the fetus parvovirus. Lack of maternal IgM antibodies in serum due to parvovirus seroconversion during pregnancy does not exclude the persistence of the virus in the fetus. To analyze the diagnostic value of the method for determining the LHP parvovirus B19 DNA in the amniotic fluid, umbilical cord blood of newborns to determine vertical transmission of parvovirus infection when infected mothers B19 during pregnancy.

  11. Comparison among the BED capture enzyme immunoassay test and AxSYM avidity index assay for determining recent HIV infection and incidence in two Voluntary Counselling and Testing Centres in Northeast Brazil

    Directory of Open Access Journals (Sweden)

    Daniela Medeiros Salustiano

    2014-07-01

    Full Text Available The aims of this study were to compare the automated AxSYM avidity assay index with the BED capture enzyme immunoassay test and to calculate the HIV-1 incidence using the BED capture enzyme immunoassay and AxSYM avidity assay index algorithms within a population seeking the Voluntary Counselling and Testing Centres in two municipalities in the Metropolitan Region of Recife, Northeast of Brazil. An analysis was conducted in 365 samples that tested positive for HIV infection from frozen serum collected during the period 2006–2009. There was a similar proportion of males and females; most patients were heterosexual (86% with a median age of 29 years. Of the 365 samples, 102 (28% and 66 (18.1% were identified as recent infections by BED capture enzyme immunoassay and AxSYM avidity assay index, respectively. The HIV-1 total incidence in the BED capture enzyme immunoassay and AxSYM avidity assay index algorithms were: 0.79 (95% CI: 0.60–0.98 and 0.34 (95% CI: −0.04 to 0.72, respectively. Incidence was higher among men. There was good agreement between the tests, with a kappa of 0.654 and a specificity of 95.8%. AxSYM avidity assay index may be helpful in improving the quality of the estimates of recent HIV infection and incidence, particularly when used in a combined algorithm with BED capture enzyme immunoassay.

  12. Fast and sensitive detection of enteropathogenic Yersinia by immunoassays.

    Science.gov (United States)

    Laporte, Jérôme; Savin, Cyril; Lamourette, Patricia; Devilliers, Karine; Volland, Hervé; Carniel, Elisabeth; Créminon, Christophe; Simon, Stéphanie

    2015-01-01

    Yersinia enterocolitica and Yersinia pseudotuberculosis, the two Yersinia species that are enteropathogenic for humans, are distributed worldwide and frequently cause diarrhea in inhabitants of temperate and cold countries. Y. enterocolitica is a major cause of foodborne disease resulting from consumption of contaminated pork meat and is further associated with substantial economic cost. However, investigation of enteropathogenic Yersinia species is infrequently performed routinely in clinical laboratories because of their specific growth characteristics, which make difficult their isolation from stool samples. Moreover, current isolation procedures are time-consuming and expensive, thus leading to underestimates of the incidence of enteric yersiniosis, inappropriate prescriptions of antibiotic treatments, and unnecessary appendectomies. The main objective of the study was to develop fast, sensitive, specific, and easy-to-use immunoassays, useful for both human and veterinary diagnosis. Monoclonal antibodies (MAbs) directed against Y. enterocolitica bioserotypes 2/O:9 and 4/O:3 and Y. pseudotuberculosis serotypes I and III were produced. Pairs of MAbs were selected by testing their specificity and affinity for enteropathogenic Yersinia and other commonly found enterobacteria. Pairs of MAbs were selected to develop highly sensitive enzyme immunoassays (EIAs) and lateral flow immunoassays (LFIs or dipsticks) convenient for the purpose of rapid diagnosis. The limit of detection of the EIAs ranged from 3.2 × 10(3) CFU/ml to 8.8 × 10(4) CFU/ml for pathogenic serotypes I and III of Y. pseudotuberculosis and pathogenic bioserotypes 2/O:9 and 4/O:3 of Y. enterocolitica and for the LFIs ranged from 10(5) CFU/ml to 10(6) CFU/ml. A similar limit of detection was observed for artificially contaminated human feces. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  13. Comparison among performances of a ligase chain reaction-based assay and two enzyme immunoassays in detecting Chlamydia trachomatis in urine specimens from men with nongonococcal urethritis.

    Science.gov (United States)

    Deguchi, T; Yasuda, M; Uno, M; Tada, K; Iwata, H; Komeda, H; Maeda, S; Latila, V; Saito, I; Kawada, Y

    1996-01-01

    We evaluated the performances of a ligase chain reaction (LCR)-based assay and two enzyme immunoassays (Chlamydiazyme and IDEIA) in the detection of Chlamydia trachomatis in urine specimens. We compared the results of testing urine specimens by these assays with those of urethral swab culture by examining samples from 131 men with nongonococcal urethritis. Discrepant results were analyzed by testing urethral swab specimens for C. trachomatis by a PCR-based assay. After the resolution of discrepant results, the sensitivity of urethral swab culture was 85.3%, whereas those of the LCR assay, Chlamydiazyme, and IDEIA with urine specimens were 94.1, 82.4, and 94.1%, respectively. The LCR assay and IDEIA were more sensitive than was urethral swab culture. In addition, the LCR assay, with a sensitivity equal to that of IDEIA, was more specific. Overall, the LCR assay proved to be superior to the enzyme immunoassays in detecting C. trachomatis in urine specimens. Testing urine specimens by LCR assay should be a helpful alternative method for diagnosing C. trachomatis urethral infection in men with nongonococcal urethritis. PMID:8784574

  14. Rapid detection and counting of viable beer-spoilage lactic acid bacteria using a monoclonal chemiluminescence enzyme immunoassay and a CCD camera.

    Science.gov (United States)

    March, Carmen; Manclús, Juan J; Abad, Antonio; Navarro, Alfonso; Montoya, Angel

    2005-08-01

    A chemiluminescence enzyme immunoassay carried out with a monoclonal antibody (MAb) and a charge-coupled device (CCD) camera was developed for rapid enumeration of viable beer-spoilage lactic acid bacteria. LA-4 MAb, which recognizes a broad spectrum of lactic acid bacteria isolated from several breweries across Spain, was produced and characterized. Test samples were filtered through polycarbonate membranes, and the membranes with retained bacteria were incubated at 31 degrees C for 2 days. They were then subjected to a two-step chemiluminescence enzyme immunoassay with MAb LA-4, and light-emitting points were detected and counted with a CCD camera. Eighteen out of 19 beer-spoilage lactic acid bacteria analysed produced luminous spots that could be enumerated. Results provided by the immunochemiluminescence assay correlated very well with those obtained by visual plate counting within a range of 3-100 CFU/100 ml. Correlation coefficients were 0.994 for four strains in sterile saline solution and 0.984 for 14 strains in artificially contaminated beer. The excellent agreement suggests that luminous spots detected within 2 days of culture are produced only by viable cells.

  15. Critical factors for EIA implementation

    DEFF Research Database (Denmark)

    Zhang, Jasmine; Kørnøv, Lone; Christensen, Per

    2013-01-01

    After decades of development, the gap between expectations of Environment Impact Assessments (EIA) and their practical performance remains significant. Research has been done to identify the critical factors for an effective implementation of EIA. However, this research, to a large extent, has...... not been cumulated and analysed comprehensively according to the stages of the EIA process. This paper contributes to the critical review of the literature on EIA implementation and effectiveness by cumulating mainly empirical findings in an implementation theoretical perspective. It focuses on the links...... between different critical factors and how they relate to different stages in the EIA and thus influence the decision making process. After reviewing 33 refereed journal articles published between 1999 and 2011, we identified 203 notions of critical factors. Of these, 102 related to different stages...

  16. Measurements in international units of antibody to hepatitis B surface antigen(anti-HBs) after immunization with a yeast-derived, subtype adr hepatitis B vaccine are considerably different between chemiluminescent immunoassay (CLIA) and chemiluminescent enzyme immunoassay (CLEIA).

    Science.gov (United States)

    Ogata, Norio

    2006-04-01

    The worldwide consensus of the minimum protective anti-HBs level against HBV infection is 10 mIU/mL on assays standardized by the World Health Organization (WHO) reference preparations. To investigate whether this value could be applied to recipients of yeast-derived recombinant HB vaccine containing the major surface protein of subtype adr (Bimmugen, Astellas Pharmaceutical, Tokyo), we compared anti-HBs measurements between chemiluminescent immunoassay (CLIA) (Architect Ausab, Abbott Japan, Tokyo) and chemiluminescent enzyme immunoassay (CLEIA) (Lumipulse Forte, Fujirebio, Tokyo) in given serum samples obtained from the vaccinees. The vaccine and the two assay methods are currently in a wide use in Japan. The study included 300 medical students who completed a standard vaccination course (0, 1 and 6 months). Serum samples obtained 1 month or 13 months after completing the vaccination were simultaneously tested for anti-HBs by CLIA and CLEIA. In 147 samples with quantifiable values on both CLIA and CLEIA (10 to 1000 mIU/mL) the geometric mean titer on CLEIA (225.0 mIU/mL) was significantly higher than that on CLIA (94.5 mIU/mL) (p < 0.0001). Of 26 subjects with CLIA measurements below 10 mIU/mL, 15 samples (57.7%) showed CLEIA measurements more than 10 mIU/mL. Thus, in the subtype adr-vaccinees CLEIA demonstrated considerably high serum anti-HBs measurements compared to CLIA and discordance in determining critical anti-HBs level of 10 mIU/mL was observed in more than half the samples. This suggests that the minimum HBV-protective anti HBs titer of 10 mIU/mL is difficult to be introduced to Japan where subtype adr-HB vaccines or -HBV infection are prevalent, unless characteristics of assay methods are carefully evaluated.

  17. Specificity of immunoassays. Pt. 2

    International Nuclear Information System (INIS)

    Pratt, J.J.; Woldring, M.G.; Boonman, R.; Kittikool, J.

    1979-01-01

    Practical aspects of the measurement of the specificity of immunoassay are reviewed. Antibody heterogeneity in an antiserum makes a pragmatic rather than a theoretical approach necessary. A new method for the measurement of immunoassay specificity is described. This method is based on the errors caused by the cross-reacting antigens and is directly relevant to the validity of results obtained by immunoassay methods. The effect of selectively blocking the least specific antibodies in antisera raised against steroid haptens is tested. The practical consequences of these considerations are tested using steroid radioimmunoassay and enzyme-immunoassay. (orig.) [de

  18. A New Enzyme Immunoassay for the Quantitative Determination of Classical Autotaxins (ATXα, ATXβ, and ATXγ and Novel Autotaxins (ATXδ and ATXε.

    Directory of Open Access Journals (Sweden)

    Yasunori Tokuhara

    Full Text Available Autotaxin (ATX is a secreted enzyme that converts lysophosphatidylcholine to lysophosphatidic acid, a potent bioactive lipid mediator, through its lysophospholipase D activity. Although five alternative splicing isoforms of ATX have been identified as ATXα, ATXβ, ATXγ, ATXδ, and ATXε and the expression patterns of each isoform differ among several tissues, the clinical significance of each isoform remains to be elucidated.Anti-ATXβ and anti-ATXδ monoclonal antibodies were produced by immunization with recombinant human ATXβ and ATXδ expressed using a baculovirus system, respectively. We then developed enzyme immunoassays to measure the serum concentrations of "classical ATX" (ATXα, ATXβ, and ATXγ and "novel ATX" (ATXδ and ATXε antigens and evaluated the usefulness of these assays using human serum samples.The with-run and between-run precision, interference, detection limit, and linearity studies for the present assay were well validated. In healthy subjects, the serum concentrations of classical ATX and novel ATX were significantly (P < 0.01 higher in women than in men, while the ratios of classical ATX or novel ATX to total ATX were not different between women and men. The concentrations of both classical ATX and novel ATX in normal pregnant subjects and patients with chronic liver diseases or follicular lymphoma were significantly higher than those in healthy subjects, while the ratio of both ATX isoforms to total ATX did not vary among these groups.We have developed a new enzyme immunoassay to determine the concentrations of classical ATX and novel ATX in human serum. These assays may be helpful for elucidating the distinct functional roles of each ATX isoform, which are largely unknown at present.

  19. A New Enzyme Immunoassay for the Quantitative Determination of Classical Autotaxins (ATXα, ATXβ, and ATXγ) and Novel Autotaxins (ATXδ and ATXε)

    Science.gov (United States)

    Tokuhara, Yasunori; Kurano, Makoto; Shimamoto, Satoshi; Igarashi, Koji; Nojiri, Takahiro; Kobayashi, Tamaki; Masuda, Akiko; Ikeda, Hitoshi; Nagamatsu, Takeshi; Fujii, Tomoyuki; Aoki, Junken; Yatomi, Yutaka

    2015-01-01

    Background Autotaxin (ATX) is a secreted enzyme that converts lysophosphatidylcholine to lysophosphatidic acid, a potent bioactive lipid mediator, through its lysophospholipase D activity. Although five alternative splicing isoforms of ATX have been identified as ATXα, ATXβ, ATXγ, ATXδ, and ATXε and the expression patterns of each isoform differ among several tissues, the clinical significance of each isoform remains to be elucidated. Methods Anti-ATXβ and anti-ATXδ monoclonal antibodies were produced by immunization with recombinant human ATXβ and ATXδ expressed using a baculovirus system, respectively. We then developed enzyme immunoassays to measure the serum concentrations of “classical ATX” (ATXα, ATXβ, and ATXγ) and “novel ATX” (ATXδ and ATXε) antigens and evaluated the usefulness of these assays using human serum samples. Results The with-run and between-run precision, interference, detection limit, and linearity studies for the present assay were well validated. In healthy subjects, the serum concentrations of classical ATX and novel ATX were significantly (P ATX or novel ATX to total ATX were not different between women and men. The concentrations of both classical ATX and novel ATX in normal pregnant subjects and patients with chronic liver diseases or follicular lymphoma were significantly higher than those in healthy subjects, while the ratio of both ATX isoforms to total ATX did not vary among these groups. Conclusions We have developed a new enzyme immunoassay to determine the concentrations of classical ATX and novel ATX in human serum. These assays may be helpful for elucidating the distinct functional roles of each ATX isoform, which are largely unknown at present. PMID:26083365

  20. Rapid detection of fungal alpha-amylase in the work environment with a lateral flow immunoassay.

    Science.gov (United States)

    Bogdanovic, Jelena; Koets, Marjo; Sander, Ingrid; Wouters, Inge; Meijster, Tim; Heederik, Dick; van Amerongen, Aart; Doekes, Gert

    2006-11-01

    Occupational allergen exposure assessment usually requires airborne dust sampling at the worksite followed by dust extraction and enzyme immunoassay (EIA) analysis at the laboratory. Use of semiquantitative lateral flow immunoassays (LFIAs) may allow a more rapid detection procedure with direct on-site demonstration of a bioallergen exposure hazard. In a field study, we evaluated a recently developed LFIA for fungal alpha-amylase, an important bakery allergen. Airborne and surface dust (wipe) samples and samples from flours and baking additives used at the workplace were collected in 5 industrial bakeries and tested in the LFIA for fungal amylase. For comparison, amylase was measured in sample eluates with the reference EIA method. Sensitivity of the LFIA was 1 to 10 ng/mL, and of EIA, approximately 25 pg/mL. In LFIA, most flour samples, 84% of wipe samples, 26% of personal airborne dust, and none of the 26 ambient air dust samples produced a visible reaction. Wipe samples from dough-making areas and flour samples gave the strongest reactions. All extracts with >5 ng allergen per milliliter showed a positive LFIA reaction. The LFIA for fungal amylase is an easy and rapid method to demonstrate the allergen directly at the worksite in less than 10 to 20 minutes. Similar LFIA methods may be used for other occupational allergens in other work environments. Lateral flow immunoassays for occupational allergens may be of great value in occupational hygiene surveys to demonstrate directly to workers and supervisors the hazards of work-related bioallergen exposure.

  1. Central nervous system blastomycosis diagnosed using the MVista® Blastomyces quantitative antigen enzyme immunoassay test on cerebrospinal fluid: A case report and review of the literature.

    Science.gov (United States)

    Walkty, Andrew; Keynan, Yoav; Karlowsky, James; Dhaliwal, Perry; Embil, John

    2018-02-01

    Blastomyces dermatitidis is a thermally dimorphic fungus that is capable of causing pulmonary and extra-pulmonary disease, including infections of the central nervous system (CNS). Diagnosis of CNS blastomycosis with non-invasive testing can be difficult, and a surgical biopsy may ultimately be required for microbiological and/or histopathological confirmation. A case of B. dermatitidis meningitis is presented where the diagnosis was made by testing cerebrospinal fluid (CSF) using the MVista® Blastomyces Quantitative Antigen Enzyme Immunoassay test. The utility of performing this test on CSF for diagnosis of CNS mass lesions/abscesses caused by B. dermatitidis in the absence of associated meningitis remains unclear. Cross reaction of the Blastomyces antigen test with other dimorphic fungi is a concern, necessitating that positive test results are interpreted in the context of the patient's exposure and travel history. Copyright © 2017 Elsevier Inc. All rights reserved.

  2. Serological discrimination by indirect enzyme immunoassay between the antibody response to Brucella sp and Yersinia enterocolitica O : 9 in cattle and pigs

    DEFF Research Database (Denmark)

    Nielsen, K.; Smith, P.; Yu, W.

    2006-01-01

    A rapid, inexpensive and rugged serological test that distinguishes cattle and swine infected with Brucella sp. or Yersinia enterocolitica O:9 is described. The test protocol, which is an indirect enzyme immunoassay uses a high concentration of divalent cation chelating agents to minimize binding...... of Y. enterocolitica O:9 antibody to rough lipopolysaccharide antigen derived from B. abortus RB51. No false positive reactions were observed when testing 100 Canadian cattle and swine without any evidence of brucellosis. The assay detected 91.6% of cattle (n = 155) and 93.5% (n = 31) of swine infected...... with Brucella sp. Sera from 58 cattle and 38 swine exposed to Y. enterocolitica O:9 were negative while only 20 sera from 121 'false positive' reactors of unspecified origin gave low level positive reactions, eliminating 84% of the false positive reactions. Crown...

  3. Development of a flow-through enzyme immunoassay and application in screening green coffee samples for ochratoxin A with confirmation by high-performance liquid chromatography.

    Science.gov (United States)

    Sibanda, L; De Saeger, S; Bauters, T G; Nelis, H J; Van Peteghem, C

    2001-10-01

    A flow-through enzyme immunoassay has been developed for the screening of green coffee bean samples for ochratoxin A (OA) and was later used in a survey on OA in green coffee from different countries. The test has a sensitivity of 8 ng/g, and calculated recoveries ranged from 70 to 89% and from 86 to 95% for spiked and naturally contaminated samples, respectively. There were no significant differences in within-day and between-day assay performance (P > 0.05). Green coffee samples (15 Arabica and 7 Robusta) received from an international coffee trader were analyzed for intrinsic fungal contamination, screened for OA, and subsequently confirmed by high-performance liquid chromatography (HPLC). All 22 samples were contaminated by fungal species of the genus Aspergillus, while Penicillium species were isolated from a mere 13.6% of the total number of samples. Isolates were tested for their ability to produce OA, and only 3.9% were positive. There was no correlation between occurrence of OA-producing isolates and levels of OA in contaminated samples. Results of the screening procedure showed that 4 of the 22 samples were contaminated with 8 ng/g or higher. The HPLC method confirmed that the OA levels ranged from 27 to 168 ng/g. A fifth sample, which was shown to be negative during screening, had an OA concentration of 4 ng/g. There were no false negatives or positives recorded, and the flow-through enzyme immunoassay results correlated with those obtained by HPLC.

  4. IgG western blot for confirmatory diagnosis of equivocal cases of toxoplasmosis by EIA-IgG and fluorescent antibody test.

    Science.gov (United States)

    Khammari, Imen; Saghrouni, Fatma; Yaacoub, Alia; Gaied Meksi, Sondoss; Ach, Hinda; Garma, Lamia; Fathallah, Akila; Ben Saïd, Moncef

    2013-08-01

    The performance values of available techniques used in serodiagnosis of toxoplasmosis are satisfactory but they raise problems of equivocal and discordant results for very low IgG titers. Recently marketed, LDBio-Toxo II IgG Western blot (IB) showed an excellent correlation with the dye test. We estimated the proportion of equivocal and discordant results between the enzyme immunoassay Platelia Toxo IgG (EIA-IgG) and fluorescent antibody test (FAT) and assessed the usefulness of the IB as a confirmatory test. Out of 2,136 sera collected from pregnant women, 1,644 (77.0%) tested unequivocally positive and 407 (19.0%) were negative in both EIA-IgG and FAT. The remaining 85 (4%) sera showed equivocal or discordant results. Among them, 73 (85.9%) were positive and 12 (14.1%) were negative in IB. Forty-one (89.1%) equivocal sera in EIA-IgG and 46 (86.8%) equivocal sera in FAT were positive in IB. Reducing the cut-off values of both screening techniques improved significantly their sensitivity in detecting very low IgG titers at the expense of their specificity. In conclusion, equivocal results in routine-used techniques and their discordance in determination of the immune status in pregnancy women were not uncommon. IB test appeard to be highly useful in these situations as a confirmatory technique.

  5. Pharmacokinetics and pharmacodynamics profiles of enalapril maleate in healthy volunteers following determination of enalapril and enalaprilat by two specific enzyme immunoassays.

    Science.gov (United States)

    Arafat, T; Awad, R; Hamad, M; Azzam, R; Al-Nasan, A; Jehanli, A; Matalka, K

    2005-08-01

    Most of the pharmacokinetic (PK) parameters for enalapril and enalaprilat were established following determination of the drug and its metabolite, using angiotensin converting enzyme (ACE) inhibition assays. In these methods, enalapril has to be hydrolysed to enalaprilat first and then assayed. The purpose of this study was to re-estimate the PK parameters of enalapril and enalaprilat in healthy volunteers using two specific enzyme immunoassays for enalapril and enalaprilat. The rate and extent of absorption of enalapril and enalaprilat from a 10-mg dose of two enalapril maleate commercial brands (Renetic and Enalapril) were estimated using a two-way-cross over design with 1-week washout period. Blood pressure was also measured at specified time intervals and correlated to enalaprilat plasma concentrations. For enalapril, the AUC(o-->infinity) values (Mean+/-SD) were 450.0+/-199.5 and 479.6+/-215.6 ng h/mL, Cmax values were 313.5+/-139.6 and 310.1+/-186.6 ng/mL, Tmax values were 1.06+/-0.30 h and 1.13+/-0.22 h, and t1/2 ranged between 0.3 to 6.1 h (1.6+/-1.5) and 0.40 to 5.05 h (1.3+/-1.0), for the two brands. For enalaprilat, the AUC(o-->infinity) values were 266.9+/-122.7 and 255.9+/-121.8 ng h/ml, Cmax values were 54.8+/-29.5 and 57.2+/-29.0 ng/mL, Tmax values were 4.6+/-1.6 h and 4.3+/-1.45 h, and t1/2 ranged between 1.1 to 10.5 h (4.5+/-2.9) and 0.6 to 9.4 h (3.5+/-2.5) for the two brands. Cmax values for enalapril are about 10 times those published in the literature and the rate and extent of absorption of the two brands of enalapril and their deesterification to enalaprilat following the administration of either brand were bioequivalent. Secondly, enalaprilat concentrations at 12-24 h following a single oral dose of enalapril in healthy volunteers were lower than those reported in the literature. The values reported here correlated with the return of blood pressure to predose level. Thirdly, enzyme immunoassays for enalapril and enalaprilat are better than

  6. An enzyme immunoassay for determining epidermal growth factor (EGF) in human serum samples using an ultramicroanalytical system.

    Science.gov (United States)

    Castells Martínez, Elisa María; Del Valle, Ruben; González, Ernesto Carlos; Melchor, Antonio; Pérez, Pedro Lucio; González, Idania; Carr, Adriana; León, Kalet

    2017-01-01

    Human epidermal growth factor is a small peptide consisting of 53 amino acid residues, which stimulates cell proliferation and is associated with several human carcinomas. A simple sandwich-type ultramicroELISA assay (UMELISA), based on the advantages of high affinity reaction between streptavidin and biotin has been developed for the measurement of EGF in human serum samples. Strips coated with a high affinity monoclonal antibody directed against EGF are used as solid phase, to ensure the specificity of the assay. The EGF assay was completed in 18 hr, with a measuring range of 39-2500 pg/mL. The intra- and inter-assay coefficients of variation were 4.4-7.3% and 0-5.1%, respectively, depending on the EGF concentrations evaluated. Percentage recovery ranged from 96-104%. Regression analysis showed a good correlation with the commercially available Human EGF Immunoassay Quantikine ® ELISA kit (n = 130, r = 0.92, P characteristics of our UMELISA EGF endorse its use for the quantification of EGF in human serum samples.

  7. A comparison of antibody capture radio- and enzyme immunoassays with immunofluorescence for detecting IgM antibody in infants with congenital rubella

    International Nuclear Information System (INIS)

    Chantler, S.; Evans, C.J.; Mortimer, P.P.; Cradock-Watson, J.E.; Ridehalgh, M.K.S.

    1982-01-01

    IgM antibody capture radioimmunoassay (MACRIA) and enzyme immunoassay (MACEIA) were compared with immunofluorescence (IF) for detecting specific IgM antibody in 99 sera from 76 infants with confirmed congenital rubella, and 61 sera from a comparative group of 59 infants who had miscellaneous abnormalities but in whom congenital rubella was not confirmed. All of 35 specimens collected from confirmed cases within 12 weeks of birth were positive by all three methods and all but one of 17 specimens collected after the age of 18 months were uniformly negative. At intermediate ages discrepancies occurred in 18 specimens, of which eight were positive and 10 negative by IF. Three of these 18 specimens were negative by both antibody capture procedures but showed weak fluorescence; the other 15 were negative by MACEIA, but positive by MACRIA which appears to be the more sensitive of the antibody capture methods. Sera from five infants in the comparative group were clearly positive by all three methods. These five infants were probably congenitally infected with rubella. Sera from the other 54 infants were negative, except for one that gave a weakly positive result by MACRIA alone. Antibody capture procedures offer several advantages over previous methods for detecting IgM antibody. Although MACRIA was found to be slightly more sensitive than MACEIA, the greater stability of the enzyme label, together with the possibility of both visual and quantitative assessment, could make MACEIA the method of choice for detecting rubella-specific IgM. (Auth.)

  8. Comparison of antibody capture radio- and enzyme immunoassays with immunofluorescence for detecting IgM antibody in infants with congenital rubella

    Energy Technology Data Exchange (ETDEWEB)

    Chantler, S.; Evans, C.J. (Wellcome Research Lab., Beckenham (UK)); Mortimer, P.P. (Virus Reference Laboratory, Central Public Health Laboratory, Colindale Avenue, London, (UK)); Cradock-Watson, J.E.; Ridehalgh, M.K.S. (Withington Hospital, Manchester (UK))

    1982-08-01

    IgM antibody capture radioimmunoassay (MACRIA) and enzyme immunoassay (MACEIA) were compared with immunofluorescence (IF) for detecting specific IgM antibody in 99 sera from 76 infants with confirmed congenital rubella, and 61 sera from a comparative group of 59 infants who had miscellaneous abnormalities but in whom congenital rubella was not confirmed. All of 35 specimens collected from confirmed cases within 12 weeks of birth were positive by all three methods and all but one of 17 specimens collected after the age of 18 months were uniformly negative. At intermediate ages discrepancies occurred in 18 specimens, of which eight were positive and 10 negative by IF. Three of these 18 specimens were negative by both antibody capture procedures but showed weak fluorescence; the other 15 were negative by MACEIA, but positive by MACRIA which appears to be the more sensitive of the antibody capture methods. Sera from five infants in the comparative group were clearly positive by all three methods. These five infants were probably congenitally infected with rubella. Sera from the other 54 infants were negative, except for one that gave a weakly positive result by MACRIA alone. Antibody capture procedures offer several advantages over previous methods for detecting IgM antibody. Although MACRIA was found to be slightly more sensitive than MACEIA, the greater stability of the enzyme label, together with the possibility of both visual and quantitative assessment, could make MACEIA the method of choice for detecting rubella-specific IgM.

  9. EQCM Immunoassay for Phosphorylated Acetylcholinesterase as a Biomarker for Organophosphate Exposures Based on Selective Zirconia Adsorption and Enzyme-Catalytic Precipitation

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Hua; Wang, Jun; Choi, Daiwon; Tang, Zhiwen; Wu, Hong; Lin, Yuehe

    2009-03-01

    A zirconia (ZrO2) adsorption-based immunoassay by electrochemical quartz crystal microbalance (EQCM) has been initially developed, aiming at the detection of phosphorylated acetylcholinesterase (AChE) as a potential biomarker for bio-monitoring exposures to organophosphate (OP) pesticides and chemical warfare agents. Hydroxyl-derivatized monolayer was preferably chosen to modify the crystal serving as the template for directing the electro-deposition of ZrO2 film with uniform nanostructures. The resulting ZrO2 film was utilized to selectively capture phosphorylated AChE from the sample media. Horseradish peroxidase (HRP)-labeled anti-AChE antibodies were further employed to recognize the captured phosphorylated protein. Enzyme-catalytic oxidation of the benzidine substrate resulted in the accumulation of insoluble product on the functionalized crystal. Ultrasensitive EQCM quantification by mass-amplified frequency responses as well as rapid qualification by visual color changes of product could be thus achieved. Moreover, 4-chloro-1-naphthol (CN) was comparably studied as an ideal chromogenic substrate for the enzyme-catalytic precipitation. Experimental results show that the developed EQCM technique can allow for the detection of phosphorylated AChE in human plasma. Such an EQCM immunosensing format opens a new door towards the development of simple, sensitive, and field-applicable biosensor for biologically monitoring low-level OP exposures.

  10. EIA and EINP. Evaluation study

    International Nuclear Information System (INIS)

    Lande, R.W.I. van der; De Vries, E.F.

    2001-01-01

    The evaluation study on the title subjects concerns two subsidy tools in the Netherlands: the Energy Investment Rebate (EIA, abbreviated in Dutch) and the Subsidy for Energy in the non-profit sector and other special sectors (EINP, abbreviated in Dutch). The central question in the evaluation was to what extent did the EIA and EINP contribute to the original policy targets and at what costs. The evaluation has been carried out by means of a desk study, interviews, and an analysis of bottlenecks and possible solutions. [nl

  11. Comparison of double-antibody radioimmunoassay with Farr-technique radioimmunoassay and double-antibody enzyme immunoassay for α-fetoprotein

    International Nuclear Information System (INIS)

    Kirkpatrick, A.; Wepsic, H.T.; Nakamura, R.M.

    1977-01-01

    We describe double-antibody procedures for determining α-fetoprotein in human serum. An equilibrium procedure can be done in 24 h with a sensitivity of at least 4 μg/liter and coefficient of variation of 5.5%. There are no interferences from normal human sera or sera with certain commonly seen chemical abnormalities. We also describe and discuss sequential procedures that range in sensitivity from 250 ng to 1 μg/liter and require 24 to 48 h incubation. The precise (mid-range) portion of the dose/response curve for sequential procedures can be shifted to higher or lower values by an adjustment of the time of preliminary incubation of antibody with unlabeled antigen. With a 37 0 C incubation, a sequential procedure can be completed in 7 h. Sensitivity is 1 μg/liter, and coefficient of variation 8.0%. The relative merits of the above assay procedures are discussed. The double-antibody radioimmunoassay is twice as sensitive as the Farr procedure and it is free of the large and variable nonspecific precipitation that accompanies the precipitation of bound antigen with sodium sulfate solution. Double-antibody radioimmunoassay is superior to enzyme immunoassay in both sensitivity and precision

  12. Development of a nanobody-alkaline phosphatase fusion protein and its application in a highly sensitive direct competitive fluorescence enzyme immunoassay for detection of ochratoxin A in cereal.

    Science.gov (United States)

    Liu, Xing; Xu, Yang; Wan, De-bin; Xiong, Yong-hua; He, Zhen-yun; Wang, Xian-xian; Gee, Shirley J; Ryu, Dojin; Hammock, Bruce D

    2015-01-20

    A rapid and sensitive direct competitive fluorescence enzyme immunoassay (dc-FEIA) for ochratoxin A (OTA) based on a nanobody (Nb)-alkaline phosphatase (AP) fusion protein was developed. The VHH (variable domain of heavy chain antibody) gene of Nb28 was subcloned into the expression vector pecan45 containing the AP double-mutant gene. The Nb28-AP construct was transformed into Escherichia coli BL21(DE3)plysS, and soluble expression in bacteria was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot. Both the Nb properties and AP enzymatic activity were validated by colorimetric and fluorometric analysis. The 50% inhibitory concentration and the detection limit of the dc-FEIA were 0.13 and 0.04 ng/mL, respectively, with a linear range of 0.06-0.43 ng/mL. This assay was compared with LC-MS/MS, and the results indicated the reliability of Nb-AP fusion protein-based dc-FEIA for monitoring OTA contamination in cereal.

  13. Bisphenol A determination in baby bottles by chemiluminescence enzyme-linked immunosorbent assay, lateral flow immunoassay and liquid chromatography tandem mass spectrometry.

    Science.gov (United States)

    Maiolini, Elisabetta; Ferri, Elida; Pitasi, Agata Laura; Montoya, Angel; Di Giovanni, Manuela; Errani, Ermanno; Girotti, Stefano

    2014-01-07

    Two immunoassays, a Lateral Flow ImmunoAssay (LFIA) based on colloidal gold nanoparticle labels and an indirect competitive chemiluminescence enzyme-linked immunosorbent assay (CL-ELISA), were developed and a high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was optimized to assess the possible release of bisphenol A (BPA, 4,4'-isopropylidenediphenol) from different plastic baby bottles treated with simulating solutions. Coating conjugate concentration, anti-BPA antibody dilution, incubation time of the primary and secondary antibodies, and tolerance to different organic solvents were optimized to obtain the best performance of the ELISA with chemiluminescent end-point detection. The influence of different buffers on LFIA performance was also evaluated. Both methods showed good repeatability (mean CV value around 13%) and sensitivity. Reproducibility tests for CL-ELISA gave a mean CV value of about 25%. The IC50 and Limit of Detection (LOD) values of CL-ELISA were 0.2 and 0.02 ng mL(-1), respectively. The LOD of LFIA was 0.1 μg mL(-1). A LC-MS/MS method was also optimized. The separation was performed in a C18 column with a triple-quadrupole mass spectrometer with electrospray ionisation interface. The method showed a good linearity in the range 2 to 500 ng mL(-1), with a regression coefficient of 0.998. In the simulating solutions the detection and quantification limits, calculated by the signal to noise level of 3 (S/N = 3), were 5.8 ng mL(-1) and 17.4 ng mL(-1), respectively. This limit of quantification was about 3 and 35 times lower than the permitted limits set by the official method CEN/TS 13130-13 (0.05 μg mL(-1)) and by the Directive 2004/19/EC (0.6 μg mL(-1)), respectively. The methods were applied to determine BPA release from baby bottles, performing repeated procedures according to EU and national regulations. The results demonstrated that no BPA migration from the tested plastic materials occurred with only one

  14. Use of a monoclonal antibody in an enzyme immunoassay for the detection of Entamoeba histolytica in fecal specimens.

    Science.gov (United States)

    Ungar, B L; Yolken, R H; Quinn, T C

    1985-05-01

    An indirect enzyme-linked immunosorbent assay (ELISA) was developed for the detection of Entamoeba histolytica in human feces, using both a monoclonal antibody and rabbit antisera. It detected from less than 1 to 57 trophozoites of 6 E. histolytica strains. Stool specimens were positive by ELISA in 18 of 22 (82%) patients with E. histolytica and in 3 of 186 (2%) of patients without demonstrable E. histolytica in their stools. The latter included one from a child living near an asymptomatic cyst carrier and another from a traveler with giardiasis who had recently taken antibiotics. One hundred eight of 183 microscopy-and ELISA-negative specimens contained other parasites including Giardia (49 specimens), Endolimax nana (24), Entamoeba coli (21), Iodamoeba butschlii (2), and Entamoeba hartmanni (1). This ELISA for E. histolytica is a simple, sensitive and specific diagnostic tool.

  15. Danish experiences on EIA of livestock projects

    DEFF Research Database (Denmark)

    Christensen, Per

    2006-01-01

    , especially when it comes to livestock projects. In an evaluation of the Danish EIA experience, we have looked more closely at how the EIA instruments function regarding livestock projects. This article addresses both the EIA process as well as the EIA screening. It is demonstrated that the EIA screening...... sustainable development. Although the provisions laid down are the same the praxis related to the field has developed at a considerable speed. In order to understand this development we have closely examined how the decisions made by the Nature Protection Board of Appeal (NPBA) have been changed and conclude...

  16. Performance of the fourth-generation Bio-Rad GS HIV Combo Ag/Ab enzyme immunoassay for diagnosis of HIV infection in Southern Africa.

    Science.gov (United States)

    Piwowar-Manning, Estelle; Fogel, Jessica M; Richardson, Paul; Wolf, Shauna; Clarke, William; Marzinke, Mark A; Fiamma, Agnès; Donnell, Deborah; Kulich, Michal; Mbwambo, Jessie K K; Richter, Linda; Gray, Glenda; Sweat, Michael; Coates, Thomas J; Eshleman, Susan H

    2015-01-01

    Fourth-generation HIV assays detect both antigen and antibody, facilitating detection of acute/early HIV infection. The Bio-Rad GS HIV Combo Ag/Ab assay (Bio-Rad Combo) is an enzyme immunoassay that simultaneously detects HIV p24 antigen and antibodies to HIV-1 and HIV-2 in serum or plasma. To evaluate the performance of the Bio-Rad Combo assay for detection of HIV infection in adults from Southern Africa. Samples were obtained from adults in Soweto and Vulindlela, South Africa and Dar es Salaam, Tanzania (300 HIV-positive samples; 300 HIV-negative samples; 12 samples from individuals previously classified as having acute/early HIV infection). The samples were tested with the Bio-Rad Combo assay. Additional testing was performed to characterize the 12 acute/early samples. All 300 HIV-positive samples were reactive using the Bio-Rad Combo assay; false positive test results were obtained for 10 (3.3%) of the HIV-negative samples (sensitivity: 100%, 95% confidence interval [CI]: 98.8-100%); specificity: 96.7%, 95% CI: 94.0-98.4%). The assay detected 10 of the 12 infections classified as acute/early. The two infections that were not detected had viral loadsHIV-negative samples were pre-screened using a different fourth-generation test. The assay also had high sensitivity for detection of acute/early infection. False-negative test results may be obtained in individuals who are virally suppressed. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Evaluation of serum galactomannan enzyme immunoassay at two different cut-offs for the diagnosis of invasive aspergillosis in patients with febrile neutropenia

    Directory of Open Access Journals (Sweden)

    Ritin Mohindra

    2017-01-01

    Full Text Available Background: Invasive aspergillosis (IA is an increasingly common and fatal opportunistic fungal infection in patients with haematological diseases. Early diagnosis is difficult as mycological culture techniques have low sensitivity and the radiological tools have low specificity. Galactomannan enzyme immunoassay (GEI detects galactomannan in the human serum with a reported sensitivity and specificity between 30% and 100%. Aims: The aim of this study was to analyse the role of GEI in diagnosis of IA in patients with febrile neutropenia and to evaluate the role of GEI in the diagnosis of IA as per the revised (2008 European Organization for Research and Treatment of Cancer–Mycoses Study Group (EORTC–MSG criteria at two different optical density (OD cut-offs of 0.5 and 1.0. Setting: This prospective study was conducted in Safdarjung Hospital, New Delhi, India. Methods: GEI testing was performed in adult patients of febrile neutropenia with evidence of IA. Results at two different OD indices (ODIs of 0.5 and 1.0 were analysed. The evaluation of the diagnostic parameter, that is, GEI was measured in terms of sensitivity, specificity and positive and negative predictive value and was validated with the revised (2008 EORTC–MSG diagnostic criteria of IA. Results: One hundred and eleven patients had evidence of IA, of which 79 patients were GEI positive when cut-off ODI was 0.5, whereas with cut-off ODI 1.0, 55 patients were GEI positive. Conclusion: ODI of 1.0 should be considered as positive while in patients with OD between 0.5 and 1.0, repeat sampling from the patient is recommended.

  18. Lateral flow immunoassay and enzyme linked immunosorbent assay as effective immunomethods for the detection of synthetic cannabinoid JWH-200 based on the newly synthesized hapten.

    Science.gov (United States)

    Fojtíková, Lucie; Šuláková, Anna; Blažková, Martina; Holubová, Barbora; Kuchař, Martin; Mikšátková, Petra; Lapčík, Oldřich; Fukal, Ladislav

    2018-01-01

    In recent years, the use of synthetic cannabinoids (SCs) as drugs of abuse has greatly increased. SCs are associated with a risk of severe poisoning or even death. Therefore, more rapid, cost effective and reliable methods are needed, especially for the screening of drivers after traffic accidents and for detailed toxicological analysis in forensic laboratories. In this study, we developed a lateral flow immunoassay (LFIA) and an enzyme linked immunosorbent assay (ELISA) for the detection of JWH-200 in oral fluids. For this purpose a new hapten was prepared using a ten-step synthetic route. The developed immuno methods are based on antibodies obtained from rabbit immunized with synthesized hapten conjugated to carrier protein. The proposed methods are highly sensitive (LOD LFIA  = 0.08 ± 0.04 ng mL -1 ; LOD ELISA  = 0.04 ± 0.02 ng mL -1 ). They were applied to the quantification of JHW-200 in spiked oral fluids. The recoveries ranged from 82 to 134% for both methods. The results correlated excellently with results obtained using UHPLC-MS/MS (R 2 LFIA  = 0.99; R 2 ELISA  = 0.99). Our developed methods could be an important tool for analyses of JWH-200 in human oral fluids. The one-step LFIA is particularly suitable for roadside and on-site monitoring due to the rapid qualitative results it delivers, while the ELISA is especially useful for laboratory quantitative analyses of positive samples captured by LFIA.

  19. Bulk immunoassays for analysis of extracellular vesicles

    NARCIS (Netherlands)

    Coumans, Frank A. W.; Gool, Elmar L.; Nieuwland, Rienk

    2017-01-01

    There is increasing clinical interest in extracellular vesicles (EV) for diagnostic and treatment purposes. This review provides an overview of bulk immunoassays to analyse EV. Western blot and enzyme-linked immunosorbent assay are still the two predominant bulk immunoassays. Recently, new assays

  20. False-positive ethyl glucuronide immunoassay screening caused by a propyl alcohol-based hand sanitizer.

    Science.gov (United States)

    Arndt, Torsten; Grüner, Joachim; Schröfel, Stefanie; Stemmerich, Karsten

    2012-11-30

    Urine ethyl glucuronide (EtG) is considered as a specific marker of recent ethanol consumption. We describe false-positive DRI(®) EIA EtG enzyme immunoassay results caused by propyl glucuronides in urine after using a propanol-based hand sanitizer. EtG screening was done with the DRI(®) EIA EtG assay (Microgenics), using a cut-off of 0.5 mg/L as recommended by the manufacturer and of 0.1 mg/L as demanded by the German Regulations for Reissuing Drivers Licenses. Confirmatory EtG analysis was done with the ClinMass(®) EtG LC-MS/MS testkit (Recipe), extended by the mass transitions 235.1→75.1, 235.1→85.1, and 235.1→113.1 for the detection of the 1- and 2-propyl glucuronides. Self-experiments were done by staff members of our lab (n=7), using 3 mL Sterillium(®) Classic Pure (30 g/100 g 1-propanol and 45 g/100 g 2-propanol) for hand sanitation every quarter of an hour for 8 h according to DIN EN 1500:2011-05 with and without an exhauster and by passive inhalation of the sanitizer vapor. Spot urine samples were taken immediately before and up to 24 h after the first sanitizer use. False-positive immunoassay results of up to 4 mg/L or 2.3 mg/g creatinine were obtained after normal use of the sanitizer and also after passive inhalation of the sanitizer vapor (up to 0.89 mg/L or 0.61 mg/g). Immunoassay results were positive even after 4-fold use of the sanitizer (up to 0.14 mg/L or 0.38 mg/g) and up to 6 h after the last sanitizer contact (maximum 0.63 mg/L and 0.33 mg/g for sanitizer users and 0.25 mg/g after passive inhalation). Spiking of EtG-free urine with 1-propyl glucuronide (Athena Environmental Sciences) between 0.05 and 10 mg/L clearly demonstrated a cross reaction of the immunoassay of approx. 10% as compared to EtG. LC-MS/MS of urines with a positive immunoassay EtG result did not show EtG signals, but distinct signals of 1-propyl glucuronide (n-propyl glucuronide) and 2-propyl glucuronide (iso-propyl glucuronide). An exhauster effectively prevented

  1. Development of a single-chain variable fragment-alkaline phosphatase fusion protein and a sensitive direct competitive chemiluminescent enzyme immunoassay for detection of ractopamine in pork

    Energy Technology Data Exchange (ETDEWEB)

    Dong Jiexian; Li Zhenfeng; Lei Hongtao; Sun Yuanming [Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642 (China); Ducancel, Frederic [CEA, iBiTec-S, Service de Pharmacologie et d' Immnoanalyse (SPI), CEA Saclay, F-91191 Gif sur Yvette (France); Xu Zhenlin [Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642 (China); Boulain, Jean-Claude [CEA, iBiTec-S, Service de Pharmacologie et d' Immnoanalyse (SPI), CEA Saclay, F-91191 Gif sur Yvette (France); Yang Jinyi; Shen Yudong [Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642 (China); Wang Hong, E-mail: gzwhongd@63.com [Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642 (China)

    2012-07-29

    Graphical abstract: Detection model of dc-CLEIA based on anti-RAC scFv-AP fusion protein. Highlights: Black-Right-Pointing-Pointer The scFv-AP fusion protein against ractopamine (RAC) was produced. Black-Right-Pointing-Pointer A dc-CLEIA for RAC was developed based on the purified scFv-AP fusion protein. Black-Right-Pointing-Pointer The sensitivity of dc-CLEIA was 10 times as sensitive as dc-ELISA for RAC. Black-Right-Pointing-Pointer Recovery tests from pork samples were studied. Black-Right-Pointing-Pointer Good accuracy was obtained. - Abstract: A rapid, sensitive chemiluminescent enzyme immunoassay (CLEIA) for ractopamine (RAC) based on a single-chain variable fragment (scFv)-alkaline phosphatase (AP) fusion protein was developed. The scFv gene was prepared by cloning the heavy- and light-chain variable region genes (V{sub H} and V{sub L}) from hybridoma cell line AC2, which secretes antibodies against RAC, and assembling V{sub H} and V{sub L} genes with a linker by means of splicing overlap extension polymerase chain reaction. The resulting scFv gene was inserted into the expression vector pLIP6/GN containing AP to produce the fusion protein in Escherichia coli strain BL21. The purified scFv-AP fusion protein was used to develop a direct competitive CLEIA (dcCLEIA) protocol for detection of RAC. The average concentration required for 50% inhibition of binding and the limit of detection of the assay were 0.25 {+-} 0.03 and 0.02 {+-} 0.004 ng mL{sup -1}, respectively, and the linear response range extended from 0.05 to 1.45 ng mL{sup -1}. The assay was 10 times as sensitive as the corresponding enzyme-linked immunosorbent assay based on the same fusion protein. Cross-reactivity studies showed that the fusion protein did not cross react with RAC analogs. DcCLEIA was used to analyze RAC spiked pork samples, and the validation was confirmed by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS). The results showed a good correlation between

  2. Paper-based enzyme-free immunoassay for rapid detection and subtyping of influenza A H1N1 and H3N2 viruses.

    Science.gov (United States)

    Lei, Kin Fong; Huang, Chia-Hao; Kuo, Rei-Lin; Chang, Cheng-Kai; Chen, Kuan-Fu; Tsao, Kuo-Chien; Tsang, Ngan-Ming

    2015-07-09

    Development of rapid screening in the ambulatory environment is the most pressing needs for the control of spread of infectious disease. Despite there are many methods to detect the immunoassay results, quantitative measurement in rapid disease screening is still a great challenge for point-of-care applications. In this work, based on the internal structural protein, i.e., nucleoprotein (NP), and outer surface glycoproteins, i.e., H1 and H3, of the influenza viruses, specific and sensitive immunoassay on paper-based platform was evaluated and confirmed. Detection and subtyping of influenza A H1N1 and H3N2 viruses found in people were demonstrated by colorimetric paper-based sandwich immunoassay. Concentration-dependent response to influenza viruses was shown and the detection limits could achieve 2.7×10(3) pfu/assay for H1 detection and 2.7×10(4) pfu/assay for H3 detection, which are within the clinical relevant level. Moreover, detection of influenza virus from infected cell lysate and clinical samples was demonstrated to further confirm the reliability of the paper-based immunoassay. The use of paper for the development of diagnostic devices has the advantages of lightweight, ease-of-use, and low cost and paper-based immunoassay is appropriate to apply for rapid screening in point-of-care applications. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. Variability in Telavancin Cross-Reactivity among Vancomycin Immunoassays

    OpenAIRE

    McConeghy, Kevin W.; Liao, Siyun; Clark, Douglas; Worboys, Philip; Barriere, Steven L.; Rodvold, Keith A.

    2014-01-01

    Telavancin is a semisynthetic lipoglycopeptide with a dual mechanism of action against Gram-positive pathogens. Two brief reports have suggested potential cross-reactivity of telavancin with the vancomycin particle-enhanced turbidometric immunoassay (PETIA). The purpose of this study was to evaluate several commercially available vancomycin immunoassays (fluorescence polarization [FPIA], enzyme-multiplied immunoassays [EMIT], PETIA, and chemiluminescent immunoassay [CMIA]) for cross-reactivit...

  4. Survey of immunoassay techniques for biological analysis

    International Nuclear Information System (INIS)

    Burtis, C.A.

    1986-10-01

    Immunoassay is a very specific, sensitive, and widely applicable analytical technique. Recent advances in genetic engineering have led to the development of monoclonal antibodies which further improves the specificity of immunoassays. Originally, radioisotopes were used to label the antigens and antibodies used in immunoassays. However, in the last decade, numerous types of immunoassays have been developed which utilize enzymes and fluorescent dyes as labels. Given the technical, safety, health, and disposal problems associated with using radioisotopes, immunoassays that utilize the enzyme and fluorescent labels are rapidly replacing those using radioisotope labels. These newer techniques are as sensitive, are easily automated, have stable reagents, and do not have a disposal problem. 6 refs., 1 fig., 2 tabs

  5. Dutch Energy Investment Allowance (EIA). Energy List for 2013; Energie-investeringsaftrek (EIA). Energielijst 2013

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2013-01-15

    The Energy Investment Allowance (EIA) is a tax system by means of which the Dutch government supports companies with investments in energy-saving equipment and renewable energy. This brochure explains the assets eligible for EIA and how the scheme works [Dutch] De Energie-investeringsaftrek (EIA) is een fiscale regeling waarmee de overheid ondersteuning biedt voor bedrijven bij investeringen in energiebesparende bedrijfsmiddelen en duurzame energie. In deze brochure wordt uitgelegd welke bedrijfsmiddelen in aanmerking komen voor EIA en hoe de regeling werkt.

  6. Biotin IgM Antibodies in Human Blood: A Previously Unknown Factor Eliciting False Results in Biotinylation-Based Immunoassays

    Science.gov (United States)

    Chen, Tingting; Hedman, Lea; Mattila, Petri S.; Jartti, Laura; Jartti, Tuomas; Ruuskanen, Olli; Söderlund-Venermo, Maria; Hedman, Klaus

    2012-01-01

    Biotin is an essential vitamin that binds streptavidin or avidin with high affinity and specificity. As biotin is a small molecule that can be linked to proteins without affecting their biological activity, biotinylation is applied widely in biochemical assays. In our laboratory, IgM enzyme immuno assays (EIAs) of µ-capture format have been set up against many viruses, using as antigen biotinylated virus like particles (VLPs) detected by horseradish peroxidase-conjugated streptavidin. We recently encountered one serum sample reacting with the biotinylated VLP but not with the unbiotinylated one, suggesting in human sera the occurrence of biotin-reactive antibodies. In the present study, we search the general population (612 serum samples from adults and 678 from children) for IgM antibodies reactive with biotin and develop an indirect EIA for quantification of their levels and assessment of their seroprevalence. These IgM antibodies were present in 3% adults regardless of age, but were rarely found in children. The adverse effects of the biotin IgM on biotinylation-based immunoassays were assessed, including four inhouse and one commercial virus IgM EIAs, showing that biotin IgM do cause false positivities. The biotin can not bind IgM and streptavidin or avidin simultaneously, suggesting that these biotin-interactive compounds compete for the common binding site. In competitive inhibition assays, the affinities of biotin IgM antibodies ranged from 2.1×10−3 to 1.7×10−4 mol/L. This is the first report on biotin antibodies found in humans, providing new information on biotinylation-based immunoassays as well as new insights into the biomedical effects of vitamins. PMID:22879954

  7. Application of a newly developed high-sensitivity HBsAg chemiluminescent enzyme immunoassay for hepatitis B patients with HBsAg seroclearance.

    Science.gov (United States)

    Shinkai, Noboru; Matsuura, Kentaro; Sugauchi, Fuminaka; Watanabe, Tsunamasa; Murakami, Shuko; Iio, Etsuko; Ogawa, Shintaro; Nojiri, Shunsuke; Joh, Takashi; Tanaka, Yasuhito

    2013-11-01

    We modified and automated a highly sensitive chemiluminescent enzyme immunoassay (CLEIA) for surface antigen (HBsAg) detection using a combination of monoclonal antibodies, each for a specific epitope of HBsAg, and by improving an earlier conjugation technique. Of 471 hepatitis B virus (HBV) carriers seen in our hospital between 2009 and 2012, 26 were HBsAg seronegative as determined by the Abbott Architect assay. The Lumipulse HBsAg-HQ assay was used to recheck those 26 patients who demonstrated seroclearance by the Abbott Architect assay. The performance of the Lumipulse HBsAg-HQ assay was compared with that of a quantitative HBsAg detection system (Abbott Architect) and the Roche Cobas TaqMan HBV DNA assay (CTM) (lower limit of detection, 2.1 log copies/ml) using blood serum samples from patients who were determined to be HBsAg seronegative by the Abbott Architect assay. Ten patients had spontaneous HBsAg loss. Of 8 patients treated with nucleotide analogues (NAs), two were HBsAg seronegative after stopping lamivudine therapy and 6 were HBsAg seronegative during entecavir therapy. Eight acute hepatitis B (AH) patients became HBsAg seronegative. Of the 26 patients, 16 were HBsAg positive by the Lumipulse HBsAg-HQ assay but negative by the Abbott Architect assay. The differences between the two assays in terms of detectable HBsAg persisted over the long term in the spontaneous loss group (median, 10 months), the NA-treated group (2.5 months), and the AH group (0.5 months). In 9 patients, the Lumipulse HBsAg-HQ assay detected HBsAg when HBV DNA was negative by the CTM assay. HBsAg was also detected by the Lumipulse HBsAg-HQ assay in 4 patients with an anti-HBs concentration of >10 mIU/ml, 3 of whom had no HBsAg escape mutations. The automatic, highly sensitive HBsAg CLEIA Lumipulse HBsAg-HQ is a convenient and precise assay for HBV monitoring.

  8. EIA modelling for CZM applications. Part 1

    Digital Repository Service at National Institute of Oceanography (India)

    Vethamony, P.; Babu, M.T.

    stream_size 12 stream_content_type text/plain stream_name Summer_Sch_EIA_Manage_Coast_Zone_2001_84.pdf.txt stream_source_info Summer_Sch_EIA_Manage_Coast_Zone_2001_84.pdf.txt Content-Encoding ISO-8859-1 Content-Type text...

  9. EIA of anthropogenic activities on marine macrobenthos

    Digital Repository Service at National Institute of Oceanography (India)

    Govindan, K.

    stream_size 12 stream_content_type text/plain stream_name Summer_Sch_EIA_Mgmt_Coastal_Zone_Trg_Manual_158.pdf.txt stream_source_info Summer_Sch_EIA_Mgmt_Coastal_Zone_Trg_Manual_158.pdf.txt Content-Encoding ISO-8859-1 Content...

  10. Danish experiences on EIA of livestock projects

    International Nuclear Information System (INIS)

    Christensen, Per

    2006-01-01

    Since its introduction into Danish planning in 1989, Environmental Impact Assessment (EIA) has been widely discussed. At the centre of the debate has been the question of whether EIA actually offered anything new and there has been a great deal of scepticism about the efficacy of the instrument, especially when it comes to livestock projects. In an evaluation of the Danish EIA experience, we have looked more closely at how the EIA instruments function regarding livestock projects. This article addresses both the EIA process as well as the EIA screening. It is demonstrated that the EIA screening in its own right is a kind of regulatory instrument. Examining the assessments made during screening more closely, we conclude that there is still some way to go in order to make the assessment broader and more holistic in accordance with the ambitions set out in the EIA directive to contribute to a more sustainable development. Although the provisions laid down are the same the praxis related to the field has developed at a considerable speed. In order to understand this development we have closely examined how the decisions made by the Nature Protection Board of Appeal (NPBA) have been changed and conclude that these changes definitely address some of the shortcomings found in the evaluation

  11. EIA data index: an abstract journal

    International Nuclear Information System (INIS)

    1980-12-01

    The individual tables, graphs, and formatted data presented in the statistical publications of the Energy Information Administration (EIA) are abstracted and indexed. Included are a complete subject index and a report number listing for all EIA publications as well as complete ordering information for these publications. The abstracts of the tables and graphs are arranged by broad subject categories (e.g., coal, petroleum, natural gas, energy analysis and modeling) with further division occurring by subcategories (e.g., reserves, drilling and production, processing). Included here are those publications and their statistical contents which were released by the EIA from its formation in October 1977 through approximately the first half of 1980. Updates will be on a semiannual basis. The EIA Data Index is a companion volume to the EIA Publications Directory: A User's Guide (DOE/EIA-0149), which provides abstracts and indexes to all EIA publications at the document level. Both of these publications are generated from the Federal Energy Data Index (FEDEX) data base which has been developed by the EIA in cooperation with the Technical Information Center of the US Department of Energy

  12. A review of EIAs on trade policy in China: Exploring the way for economic policy EIAs

    International Nuclear Information System (INIS)

    Mao, Xianqiang; Song, Peng; Kørnøv, Lone; Corsetti, Gabriel

    2015-01-01

    During the discussion on the “Environmental Protection Law Amendment (draft)” in 2011, it was decided to drop the proposed clauses related to environmental impact assessments (EIAs) on policy, which means that there remained no provisions for policy EIAs, and China's strategic environmental assessment system stayed limited to the planning level. However, considering that economic policy making is causing significant direct and indirect environmental problems and that almost every aspect of governmental policy has an economic aspect, EIAs on economic policies are of the utmost urgency. The purpose of this study is to review the EIA work that has been carried out on trade policy in China through four case studies, and illustrate how trade policy EIAs can be helpful in achieving better environmental outcomes in the area of trade. Through the trade policy EIA case studies we try to argue for the feasibility of conducting EIAs on economic policies in China. We also discuss the implications of the case studies from the point of view of how to proceed with EIAs on economic policy and how to promote their practice. - Highlights: • SEA system is incomplete and stays limited to the plan EIA level in China. • EIA on economic policy is of utmost importance for all the developing countries. • Four case studies of trade policy EIA in China are reviewed for policy implications. • Departmental competition for political power impedes economic policy EIAs in China. • Legislative regulation on policy EIA is the first thing needed to overcome barrier

  13. Enzyme

    Science.gov (United States)

    Enzymes are complex proteins that cause a specific chemical change in all parts of the body. For ... use them. Blood clotting is another example of enzymes at work. Enzymes are needed for all body ...

  14. EIA projections of coal supply and demand

    International Nuclear Information System (INIS)

    Klein, D.E.

    1989-01-01

    Contents of this report include: EIA projections of coal supply and demand which covers forecasted coal supply and transportation, forecasted coal demand by consuming sector, and forecasted coal demand by the electric utility sector; and policy discussion

  15. EIA completes corrections to drilling estimates series

    International Nuclear Information System (INIS)

    Trapmann, W.; Shambaugh, P.

    1998-01-01

    The Energy Information Administration (EIA) has published monthly and annual estimates of US oil and gas drilling activity since 1978. These data are key information for many industry analysts, serving as a leading indicator of trends in the industry and a barometer of general industry status. They are assessed directly for trends, as well as in combination with other measures to assess the productivity and profitability of upstream industry operations. They are major reference points for federal and state policymakers. EIA does not itself collect drilling activity data. Instead, it relies on an external source for data on oil, bas, and dry well completions. These data are provided to EIA monthly on an as reported basis. During a recent effort to enhance EIA's well completion data system, the detection of unusual patterns in the well completion data as received led to an expanded examination of these data. Substantial discrepancies between the data as received by EIA and correct record counts since 1987 were identified. For total wells by year, the errors ranged up to more than 2,300 wells, 11% of the 1995 total, and the impact of these errors extended backward in time to at least the early 1980s. When the magnitude and extent of the as reported well completion data problem were confirmed, EIA suspended its publication and distribution of updated drilling data. EIA staff proceeded to acquire replacement files with the as reported records and then revise the statistical portion of its drilling data system to reflect the new information. The replacement files unfortunately also included erroneous data based on the improper allocation of wells between exploration and development. EIA has now resolved the two data problems and generated revised time series estimates for well completions and footage drilled. The paper describes the problems in the data, differences between the series, and maintaining future data quality

  16. A conventional chemical reaction for use in an unconventional assay: A colorimetric immunoassay for aflatoxin B1 by using enzyme-responsive just-in-time generation of a MnO2 based nanocatalyst.

    Science.gov (United States)

    Lai, Wenqiang; Zeng, Qiao; Tang, Juan; Zhang, Maosheng; Tang, Dianping

    2018-01-10

    The authors describe a colorimetric immunoassay for the model nalyte aflatoxin B 1 (AFB 1 ). It is based on the just-in-time generation of an MnO 2 nanocatalyst. Unlike previously developed immunoassay, the chromogenic reaction relies on the just-in-time formation of an oxidase mimic without the aid of the substrate. Potassium permanganate (KMnO 4 ) is converted into manganese dioxide (MnO 2 ) which acts as an oxidase mimic that catalyzes the oxidation 3,3',5,5'-tetramethylbenzidine (TMB) by oxygen to give a blue colored product. In the presence of ascorbic acid (AA), KMnO 4 is reduced to Mn(II) ions. This results in a decrease in the amount of MnO 2 nanocatalyst. Hence, the oxidation of TMB does not take place. By adding ascorbate oxidase, AA is converted into dehydroascorbic acid which cannot reduce KMnO 4 . Based on these observations, a colorimetric competitive enzyme immunoassay was developed where ascorbate oxidase and gold nanoparticle-labeled antibody against AFB 1 and magnetic beads carrying bovine serum albumin conjugated to AFB 1 are used for the determination of AFB 1 . In presence of AFB 1 , it will compete with the BSA-conjugated AFB 1 (on the magnetic beads) for the labeled antibody against AFB 1 on the gold nanoparticles. This makes the amount of ascorbate oxidase/anti-AFB 1 antibody-labeled gold nanoparticles, which conjugated on magnetic beads, reduce, and resulted in an increase of ascorbic acid. Under optimal conditions, the absorbance (measured at 652 nm) decreases with increasing AFB 1 concentrations in the range from 0.1 to 100 ng mL -1 , with a 0.1 ng mL -1 detection limit (at the 3S blank level). The accuracy of the assay was validated by analyzing spiked peanut samples. The results matched well with those obtained with a commercial ELISA kit. Conceivably, the method is not limited to aflatoxins but has a wide scope in that it may be applied to many other analytes for which respective antibodies are available. Graphical abstract

  17. A sensitive progesterone enzyme immunoassay for cow, goat and llama plasma using a monoclonal antibody and Danazol (17-α-2,4-pregnadien-20-yno (2,3-D) isoxazol-17-ol) as a displacing agent

    International Nuclear Information System (INIS)

    Aba, M.A.; Carlsson, M.A.; Karlsson, A.; Forsberg, M.

    2001-01-01

    A sensitive progesterone enzyme immunoassay was developed for cow, goat and llama plasma using a monoclonal antibody and Danazol (17-α-2,4-pregnadien-20-yno (2,3-d) isoxazol-17-ol) as a displacing agent. The microtitration plates were first coated with progesterone 3 (o-carboxy-methyl) oxine: BSA conjugate. The immune reaction was performed by incubating overnight a mixture of 50 μL of plasma and 100 μL of first antibody. After washing, 100 μL of the second antibody (horse radish peroxidase conjugated anti-mouse IgG) were added. The plates were incubated for 1 hour and washed. Immediately the substrate solution was added and finally the reaction stopped and optical density measured. This assay allows accurate determination of progesterone in plasma from several species with good specificity, precision and accuracy, and is suitable for the rapid assessment of luteal function and reproductive status in both clinical and research situations. (author)

  18. Ecological Input Assessment and EIA: A Study On EIA Report For Quarry Projects

    Directory of Open Access Journals (Sweden)

    Rahimah Wahid

    2016-12-01

    Full Text Available The Environmental Impact Assessment (EIA was introduced as mandatory in Malaysia since 1988 as a measurement tool to achieve sustainable development. This study attempts to assess the EIA reports for quarrying activities which have been submitted to the Department of Environment. There are 19 scheduled activities requiring an EIA prior to project implementation. As the ecological input is an important part of the EIA report, these studies have tried to analyze the ecological quality of input in four EIA reports prepared for quarrying activities in Peninsular Malaysia. The results show that all aspects of the report comply with the review and sampling methods although there are deficiencies which may be remedied. Four EIA reports show weaknesses in the acquisition of the latest available data. For the preparation of the existing environment sections, the consultants do not feel inclined to make sampling on rock habitat, aquatic habitats and flora and fauna. The insulation measures suggested are not only exhaustive but also ineffective. The residual impact on the ecology is briefly mentioned in all the reports. Generally these EIA reports are found to have many shortcomings in the quality of ecological input and can be improved. Thus, a suggestion with a comprehensive approach and the cooperation of all parties is needed to sustain and complement the EIA.

  19. Climate change in EIA - Inspiration from practice

    DEFF Research Database (Denmark)

    Larsen, Sanne Vammen

    2013-01-01

    Climate change integration has been a topic of much interest in the field of impact assessment for a period, and thus far quite some emphasis has been put on discussions of purpose, relevance and overall approaches in both Environmental Impact Assessment of projects (EIA) and Strategic...... Environmental Assessments of plans and programmes (SEA). However, EIAs and SEAs are already being made, which integrate climate change, and for some aspects this practice has evolved over a long period. This paper seeks to explore this practice and find inspiration from the work with climate change already...

  20. Padronização de um teste imunoenzimático para detecção de Salmonella em alimentos Standardization of an enzyme immunoassay for detection of Salmonella in foods

    Directory of Open Access Journals (Sweden)

    Regina Baptista dos Reis

    2002-08-01

    Full Text Available A metodologia convencional utilizada para detecção de Salmonella em alimentos é trabalhosa, apresenta custo elevado e os resultados definitivos somente estão disponíveis após 96 horas. Vários métodos rápidos têm sido propostos, sendo os testes imunoenzimáticos os mais empregados. Este estudo relata o desenvolvimento de um teste imunoenzimático para detecção de Salmonella em alimentos, empregando-se um anti-soro policlonal monovalente contendo aglutininas f,g,s, não absorvido, e um anti-soro polivalente absorvido contendo as aglutininas e,h; 1,6; i; 1,2; f,g,s e m,t. A eficiência foi comparada com a da metodologia de cultivo tradicional. O teste imunoenzimático foi empregado para a detecção de Salmonella em amostras de alimentos infantis experimentalmente inoculadas com este patógeno e com outras enterobactérias, em diferentes proporções. O teste imunoenzimático revelou-se significativamente mais sensível que o método de cultivo. Esse mesmo teste, utilizando-se o anti-soro f,g,s não absorvido com antígenos heterólogos revelou concordância de 89,6% com o método de cultivo e sensibilidade de 100,0%. Por outro lado, empregando-se o anti-soro polivalente absorvido, a concordância com o método de cultivo foi de 81,3% embora a sensibilidade tenha se mantido no mesmo nível. O desempenho do teste imunoenzimático empregando-se um desses dois anti-soros indica um grande potencial de aplicação como método de triagem na detecção de Salmonella em alimentos.The conventional method for detection of Salmonella in foods is cumbersome, it is not cost-effective and results are available only after 96h. Many alternative rapid methods have been already proposed and enzyme immunoassays are the most common. This study reports the standardization of a new enzyme immunoassay for detection of Salmonella in foods, based on a policlonal non-absorbed antiserum containing f,g,s aglutinins and a pool of policlonal absorbed antisera

  1. Comparison of Surface Plasmon Resonance, Resonant Waveguide Grating Biosensing and Enzyme Linked Immunosorbent Assay (ELISA in the Evaluation of a Dengue Virus Immunoassay

    Directory of Open Access Journals (Sweden)

    Joe Buechler

    2013-07-01

    Full Text Available Two label-free biosensor platforms, Resonance Waveguide Grating (RWG and Surface Plasmon Resonance (SPR, were used to rank a large panel of anti-dengue virus NS1 antibodies. Dengue non-structural 1 (NS1 protein is an established serological marker for the early detection of dengue infection. A variety of commercial dengue NS1 antigen capture immunoassays are available in both ELISA and lateral flow format. However, there is a significant scope to improve both the sensitivity and the specificity of those tests. The interactions of antibody (Ab-antigen (Ag were profiled, with weak interactions (KD = 1–0.1 μM able to be detected under static equilibrium conditions by RWG, but not observed to under more rigorous flow conditions using SPR. There were significant differences in the absolute affinities determined by the two technologies, and there was a poor correlation between antibodies best ranked by RWG and the lower limit of detection (LLOD found by ELISA. Hence, whilst high-throughput RWG can be useful as preliminary screening for higher affinity antibodies, care should be exercised in the assignation of quantitative values for affinity between different assay formats.

  2. EIA for a waste incinerator in Denmark

    DEFF Research Database (Denmark)

    Larsen, Sanne Vammen

    2017-01-01

    A planned new waste incinerator will be located in an area which is at risk of flooding – a risk that will increase under climate change. During public hear- ings as part of the project’s EIA, inclusion of climate risks was requested. This led to mitigation measures which will decrease the risk...

  3. Recently released EIA report presents international forecasting data

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1995-05-01

    This report presents information from the Energy Information Administration (EIA). Articles are included on international energy forecasting data, data on the use of home appliances, gasoline prices, household energy use, and EIA information products and dissemination avenues.

  4. Is EIA part of the wind power planning problem?

    International Nuclear Information System (INIS)

    Smart, Duncan Ewan; Stojanovic, Timothy A.; Warren, Charles R.

    2014-01-01

    This research evaluates the importance and effectiveness of Environmental Impact Assessment (EIA) within wind farm planning debates, drawing on insights from case studies in Scotland. Despite general public support for renewable energy on the grounds that it is needed to tackle climate change and implement sustainable development, many proposed wind farms encounter significant resistance. The importance of planning issues and (EIA) processes has arguably been overlooked within recent wind farm social acceptability discourse. Through semi-structured interviews with key stakeholders and textual analysis of EIA documents, the characteristics of EIA are assessed in terms of its perceived purpose and performance. The data show that whilst respondents perceive EIA to be important, they express concerns about bias and about the inability of EIA to address climate change and wind farm decommissioning issues adequately. Furthermore, the research identifies key issues which impede the effectiveness of EIA, and reveals differences between theoretical and practical framings of EIA. The paper questions the assumption that EIA is a universally applicable tool, and argues that its effectiveness should be analysed in the context of specific development sectors. The article concludes by reviewing whether the recently amended EIA Directive (2014/52/EU) could resolve identified problems within national EIA practice. - Highlights: • Evaluation of EIA for onshore wind farm planning in Scotland. • EIA is important for multiple aspects of onshore wind farm planning. • Multiple substantive deficiencies of relevance to wind farm planning exist in EIA. • Further research into EIA effectiveness for specific development types is required. • Directive 2014/52/EU may improve EIA effectiveness within wind farm planning

  5. Dutch Energy Investment Allowance (EIA). Annual report 2012; Energie-Investeringsaftrek (EIA). Jaarverslag 2012

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2013-07-15

    By means of the Energy Investment Allowance (EIA) the Ministry of Economic Affairs supports investments of businesses, industrial associations and other parties in accelerating innovative, energy saving sustainable initiatives and technologies. In this report the results for 2012 are presented [Dutch] Met de EIA ondersteunt het Ministerie van Economische Zaken bedrijven bij het investeren in innovatieve, energiebesparende en duurzame technieken. In dit verslag over 2012 worden resultaten weergegeven.

  6. A New Enzyme Immunoassay for the Quantitative Determination of Classical Autotaxins (ATX?, ATX?, and ATX?) and Novel Autotaxins (ATX? and ATX?)

    OpenAIRE

    Tokuhara, Yasunori; Kurano, Makoto; Shimamoto, Satoshi; Igarashi, Koji; Nojiri, Takahiro; Kobayashi, Tamaki; Masuda, Akiko; Ikeda, Hitoshi; Nagamatsu, Takeshi; Fujii, Tomoyuki; Aoki, Junken; Yatomi, Yutaka

    2015-01-01

    Background Autotaxin (ATX) is a secreted enzyme that converts lysophosphatidylcholine to lysophosphatidic acid, a potent bioactive lipid mediator, through its lysophospholipase D activity. Although five alternative splicing isoforms of ATX have been identified as ATX?, ATX?, ATX?, ATX?, and ATX? and the expression patterns of each isoform differ among several tissues, the clinical significance of each isoform remains to be elucidated. Methods Anti-ATX? and anti-ATX? monoclonal antibodies were...

  7. Immunoassays in Biotechnology

    Science.gov (United States)

    Immunoassays have broad applications for a wide variety of important biological compounds and environmental contaminants. Immunoassays can detect the presence of an antigen in the human body, a pollutant in the environment, or a critical antibody in a patient’s serum to develop a...

  8. An Assessment of Environmental Impacts Assessment (EIA in Malaysia

    Directory of Open Access Journals (Sweden)

    Abdul Hamid Masdiah

    2017-01-01

    Full Text Available The effectiveness of Environmental Impact Assessment (EIA in evaluating the planning project is a debatable issue among academics and practitioners, since EIA has been claimed to be unable to eliminate the environmental issues. Focusing only on technical improvements is not sufficient for rectifying the problems of EIA; the process of EIA should be clearly identified instead to maximise the effective use of EIA. It is important to note that the effective use of EIA, particularly on process-related issues could significantly minimise bad environmental effects. In summary, this study aims to explore and identify the effectiveness of EIA in the planning process and barriers to evaluate the environmental performance in Malaysia. The findings of this study could be a baseline for organisation to minimize emission, avoid the risk of prosecution and fines arising from potential environment breaches and cost reduction within the organisation.

  9. Discrimination of methicillin-resistant Staphylococcus aureus from methicillin-susceptible Staphylococcus aureus or coagulase-negative staphylococci by detection of penicillin-binding protein 2 and penicillin-binding protein 2' using a bioluminescent enzyme immunoassay.

    Science.gov (United States)

    Shiga, Kazuki; Gomi, Keiko; Nishimura, Motoi; Watanabe, Masaharu; Nomura, Fumio; Kajiyama, Naoki

    2013-02-28

    For the discrimination of methicillin-resistant S. aureus (MRSA) from methicillin-susceptible S. aureus (MSSA) or coagulase-negative staphylococci (CNS), we developed a bioluminescent enzyme immunoassay (BLEIA) for detecting penicillin-binding protein 2 (PBP2) and penicillin-binding protein 2' (PBP2') using biotinylated firefly luciferase. The BLEIA was able to detect recombinant PBP2 at 50 pg/ml and recombinant PBP2' at 500 pg/ml. PBP2 and PBP2' present in the membranes of S. aureus were extracted by acid and detergent treatment. The method was able to detect PBP2 or PBP2' extracted from 10(6) colony forming units of S. aureus because of efficient extraction and the high sensitivity of luciferase. In a study of clinical isolates previously characterized as either MRSA or MSSA by antibiotic susceptibility testing, all 34 specimens identified as MRSA were both PBP2 and PBP2' positive. The 34 MSSA specimens were PBP2 positive and PBP2' negative. Moreover, the BLEIA could detect PBP2' extracted from four species of methicillin-resistant CNS, but not PBP2 extracted from four species of methicillin-resistant and methicillin-susceptible CNS. This result suggested that PBP2 could be a unique marker for discrimination of S. aureus from CNS. A BLEIA that is able to detect PBP2 and PBP2' may be useful in clinical diagnostics. Copyright © 2012 Elsevier B.V. All rights reserved.

  10. Inclusion bodies of recombinant Epstein-Barr virus capsid antigen p18 as potential immobilized antigens in enzyme immunoassays for detection of nasopharyngeal carcinoma.

    Science.gov (United States)

    Lim, Chun Shen; Goh, Siang Ling; Kariapper, Leena; Krishnan, Gopala; Lim, Yat-Yuen; Ng, Ching Ching

    2015-08-25

    Development of indirect enzyme-linked immunosorbent assays (ELISAs) often utilizes synthetic peptides or recombinant proteins from Escherichia coli as immobilized antigens. Because inclusion bodies (IBs) formed during recombinant protein expression in E. coli are commonly thought as misfolded aggregates, only refolded proteins from IBs are used to develop new or in-house diagnostic assays. However, the promising utilities of IBs as nanomaterials and immobilized enzymes as shown in recent studies have led us to explore the potential use of IBs of recombinant Epstein-Barr virus viral capsid antigen p18 (VCA p18) as immobilized antigens in ELISAs for serologic detection of nasopharyngeal carcinoma (NPC). Thioredoxin fusion VCA p18 (VCA-Trx) and IBs of VCA p18 without fusion tags (VCA-IBs) were purified from E. coli. The diagnostic performances of IgG/VCA-IBs, IgG/VCA-Denat-IBs (using VCA-IBs coated in 8mol/l urea), IgG/VCA-Trx, and IgG/VCA-Peptide assays were compared by screening 100 NPC case-control pairs. The IgG/VCA-Denat-IBs assay showed the best area under the receiver operating characteristic curve (AUC: 0.802; p<0.05), while the AUCs for the IgG/VCA-IBs, IgG/VCA-Trx, and IgG/VCA-Peptide assays were comparable (AUC: 0.740, 0.727, and 0.741, respectively). We improved the diagnostic performance of the ELISA significantly using IBs of recombinant VCA p18. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. Bulk immunoassays for analysis of extracellular vesicles.

    Science.gov (United States)

    Coumans, Frank A W; Gool, Elmar L; Nieuwland, Rienk

    2017-05-01

    There is increasing clinical interest in extracellular vesicles (EV) for diagnostic and treatment purposes. This review provides an overview of bulk immunoassays to analyse EV. Western blot and enzyme-linked immunosorbent assay are still the two predominant bulk immunoassays. Recently, new assays have become available that can detect exposure to EV concentrations that are up to 10,000-fold lower. This is advantageous for applications that detect rare EV. Other important parameters are the detectable concentration range, the required sample volume, whether simultaneous presence of different antigens on a single EV can be detected, size selectivity of each assay and practical considerations. In this review, we will explain the working principles of the traditional and novel assays together with their performance parameters. The most sensitive assays are micro-nuclear magnetic resonance, surface plasmon resonance, and time-resolved fluorescent immunoassay.

  12. EIA and green procurement: Opportunities for strengthening their coordination

    International Nuclear Information System (INIS)

    Uttam, Kedar; Faith-Ell, Charlotta; Balfors, Berit

    2012-01-01

    EIA plays an important role in enhancing the environmental performance of the construction sector. In recent years, the construction sector has been developing green procurement practices. Green procurement is a process that involves the incorporation of environmental requirements during the procurement of services and products. However, discussion on green procurement is rarely seen during the EIA phase. This paper addresses possible opportunities for improving the coordination between EIA and green procurement within the construction sector. The linking of EIA and green procurement has been postulated in the paper as an aid to strengthen the coordination between project planning and implementation. The paper is based on a literature review and is an outcome of an on-going research project concerning EIA and green procurement. This study indicated that it would be appropriate to introduce green procurement during the pre-decision phase of an EIA. In the present study, the opportunities for integrating green procurement at the stage of EIA are associated with the integration of project planning and EIA. Future research should investigate the mechanism through which the link can be established. - Highlights: ► This paper identifies opportunities to link EIA and green procurement. ► Pre-decision phase of EIA could be appropriate for planning green procurement. ► Future research should investigate the mechanism for establishing the link.

  13. Enzyme immunoassay and proteomic characterization of troponin I as a marker of mammalian muscle compounds in raw meat and some meat products.

    Science.gov (United States)

    Zvereva, Elena A; Kovalev, Leonid I; Ivanov, Alexei V; Kovaleva, Marina A; Zherdev, Anatoly V; Shishkin, Sergey S; Lisitsyn, Andrey B; Chernukha, Irina M; Dzantiev, Boris B

    2015-07-01

    The skeletal muscle protein troponin I (TnI) has been characterized as a potential thermally stable and species-specific biomarker of mammalian muscle tissues in raw meat and meat products. This study proposed a technique for the quantification of TnI comprising protein extraction and sandwich enzyme-linked immunosorbent assay (ELISA). The technique is characterized by a TnI detection limit of 4.8 ng/ml with quantifiable concentrations ranging from 8.7 to 52 ng/ml. The method was shown to be suitable for detection of TnI in mammalian (beef, pork, lamb, and horse) meat but not in poultry (chicken, turkey, and duck) meat. In particular, the TnI content in beef was 0.40 3 ± 0.058 mg/g of wet tissue. The TnI estimations obtained for the pork and beef samples using ELISA were comparable to the proteomic analysis results. Thus, the quantitative study of TnI can be a convenient way to assess the mammalian muscle tissue content of various meat products. Copyright © 2015. Published by Elsevier Ltd.

  14. Solid-phase enzyme immunoassay or radioimmunoassay for the detection of immune complexes based on their recognition by conglutinin: conglutinin-binding test

    International Nuclear Information System (INIS)

    Casali, P.; Bossus, A.; Carpentier, N.A.; Lambert, P.H.

    1977-01-01

    Bovine conglutinin was used in a solid-phase assay for the detection of immune complexes. In a first step, the tested serum sample was incubated in polypropylene tubes coated with conglutinin to allow C3-coated immune complexes to bind to solid-phase conglutinin. In a second step, the conglutinin-bound complexes were detected using an enzyme-conjugated or radiolabelled anti-immunoglobulin antibody. The conglutinin-binding (KgB) test did not suffer from the interference of DNA, heparin or endotoxins. Its limit of sensitivity for aggregated IgG was 3 μg/ml undiluted human serum. Immune complexes prepared in vitro using tetanus toxoid, or DNA, and corresponding antibodies in human sera could be detected at various antigen/antibody ratios and at antibody concentrations lower than 8 μg/ml. The KgB test allowed for the detection of immune complexes in sera from patients with systemic lupus erythematosus, rheumatoid arthritis, idiopathic vasculitis, leprosy and leukemia. These sera were also tested using the 125 I-labelled Clq-binding activity (BA) test and the KgB test simultaneously, and a significant rank order correlation was observed. In patients with leukemia, a significant correlation was observed using three tests, KgB, 125 I-labelled Clq BA and Raji-cell radioimmunoassay (RIA). Therefore, the KgB test appears as a simple and reproducible method, utilizing a very stable reagent, with a sensitivity and specificity comparable to the other tests studied and allowing for clinical application. (author)

  15. EIA screening and nature protection in Denmark

    DEFF Research Database (Denmark)

    Christensen, Per; Kørnøv, Lone

    2011-01-01

    focus on the protection of groundwater and Natura 2000 sites. In particular, the implementation of Natura 2000 in Danish legislation has increased the demands on many farms. In its rulings on appealed cases, the Nature Protection Board of Appeal has strengthened its demands, and this is mirrored...... in screening practices. In this paper, the demands formulated in the guidelines of local authorities were analysed in order to investigate how the protection of groundwater, coastal waters, lakes and Natura 2000 sites develops through EIA screening. It is concluded that the level of protection has improved...

  16. The EIA mixes a US gas cocktail

    International Nuclear Information System (INIS)

    Gurney, Judith

    1999-01-01

    This article examines the US Energy Information Administration's (EIA) assumption that although gas consumption is rising there will still be adequate supplies to meet the demand in the year 2020. The expected increase in the use of gas for electricity generation, and the meeting of future demand by the expected growth in gas production in the deepwaters off the Gulf of Mexico, imports from Canada, and use of unconventional sources such as coalbed methane and natural gas hydrates are discussed, and methods of producing hydrates are outlined. US natural gas production for 1970-1998, US demand and Canadian supplies (1980-1996), and US natural gas hydrate resources are tabulated. (UK)

  17. Rough lipopolysaccharide of Brucella abortus RB51 as a common antigen for serological detection of B. ovis, B. canis, and B. abortus RB51 exposure using indirect enzyme immunoassay and fluorescence polarization assay.

    Science.gov (United States)

    Nielsen, K; Smith, P; Conde, S; Draghi de Benitez, G; Gall, D; Halbert, G; Kenny, K; Massengill, C; Muenks, Q; Rojas, X; Perez, B; Samartino, L; Silva, P; Tollersrud, T; Jolley, M

    2004-01-01

    Rough lipopolysaccharide (RLPS) antigens were prepared from cultures of Brucella abortus RB51, B. ovis, and B. canis. The preparations were standardized by weight and tested with sera from cattle immunized with B. abortus RB51, sheep infected with B. ovis, and dogs infected with B. canis. Populations of unexposed animals of each species were also tested. The tests used were the indirect enzyme immunoassay (IELISA) using RLPS and the fluorescence polarization assay (FPA) using RLPS core fractions, labeled with fluorescein isothiocyanate. The IELISA using B. abortus RB51 RLPS antigen resulted in sensitivity and specificity values of 94.8% and 97.3%, respectively, when testing bovine sera, 98.5% and 97.8% when testing ovine sera, and 95.8% and 100% when testing dog sera. The IELISA using B. ovis RLPS antigen gave sensitivity and specificity values of 80.5% and 91.7%, respectively with bovine sera, 98.9% and 93.8% with sheep sera, and 70.8% and 79.8% with dog sera. The IELISA using B. canis RLPS antigen resulted in sensitivity and specificity values of 97.0% and 97.4%, respectively, with bovine sera, 96.2% and 96.3% with sheep sera, and 95.8% and 98.8% with dog sera. Labeling RLPS core from B. ovis and B. canis with fluorescein was not successful. B. abortus RB51 core labeled with fluorescein resulted in sensitivity and specificity values of 93.5% and 99.8%, respectively, with bovine sera and 78.1% and 99.0% with sheep sera. It was not possible to test the dog sera in the FPA.

  18. Hydrogel nanoparticle based immunoassay

    Science.gov (United States)

    Liotta, Lance A; Luchini, Alessandra; Petricoin, Emanuel F; Espina, Virginia

    2015-04-21

    An immunoassay device incorporating porous polymeric capture nanoparticles within either the sample collection vessel or pre-impregnated into a porous substratum within fluid flow path of the analytical device is presented. This incorporation of capture particles within the immunoassay device improves sensitivity while removing the requirement for pre-processing of samples prior to loading the immunoassay device. A preferred embodiment is coreshell bait containing capture nanoparticles which perform three functions in one step, in solution: a) molecular size sieving, b) target analyte sequestration and concentration, and c) protection from degradation. The polymeric matrix of the capture particles may be made of co-polymeric materials having a structural monomer and an affinity monomer, the affinity monomer having properties that attract the analyte to the capture particle. This device is useful for point of care diagnostic assays for biomedical applications and as field deployable assays for environmental, pathogen and chemical or biological threat identification.

  19. Environmental Impact Assessment (Eia) On Project Design: An ...

    African Journals Online (AJOL)

    Environmental Impact Assessment (Eia) On Project Design: An Emperical Approach. ... West African Journal of Industrial and Academic Research ... Also, four key effects of EIA exercise on project design were established, such as development of design alternatives, identification of risk(s) in the design, and design recap or ...

  20. EIA modelling for coastal zone management. Part 2

    Digital Repository Service at National Institute of Oceanography (India)

    Babu, M.T.; Vethamony, P.

    stream_size 15 stream_content_type text/plain stream_name Summer_Sch_EIA_Manage_Coast_Zone_2001_95.pdf.txt stream_source_info Summer_Sch_EIA_Manage_Coast_Zone_2001_95.pdf.txt Content-Encoding ISO-8859-1 Content-Type text...

  1. EIA systems in Nigeria: evolution, current practice and shortcomings

    International Nuclear Information System (INIS)

    Ogunba, Olusegun A.

    2004-01-01

    Amidst mounting criticism of Environmental Impact Assessments (EIAs) carried out in Nigeria under the three independent EIA systems--the EIA Decree 86 (1992), the Town and Country Planning Decree 88 (1992) and the Petroleum Act (1969)--the paper traces the evolution of Nigeria's systems and appraises current practice and shortcomings. The path of development of the systems was traced within the framework of Gibson's model of EIA evolution [Impact Assess. Proj. Apprais., 20 (3) 2002, 151-159], while current practice and shortcomings were explored in random interview surveys of consultant firms, approval authorities and the academia. It was seen that Gibson's four-stage model is not exactly representative of the Nigerian situation, and a more appropriate six-stage model was developed. It was also established that the current practices of the three EIA systems were at different stages of evolution: one of the EIA schemes (the Town and Country Planning Decree) has not evolved satisfactorily, while the other two EIA systems have produced intricate legislations and guidelines, but fall short of first-rate practice. The other discovery was that the simultaneous use of three independent systems creates unnecessary duplication of EIA preparation with considerable time and money costs. The paper advises that Nigeria can make substantial progress along the evolutionary path through a correction of observed system shortcomings and a merger of the three systems

  2. EIA-CES Rehabilitation Engineering Seedbed

    Directory of Open Access Journals (Sweden)

    Juliana Velásquez Gómez

    2016-12-01

    Full Text Available La discapacidad es una interacción entre los estados de salud (enfermedades, lesiones, entre otros y los factores contextuales (factores ambientales y personales. En el semillero de Ingeniería de Rehabilitación de la Universidad EIA se evalúan usuarios en situación de discapacidad para realizarles productos de apoyo a la medida. Este semillero es voluntario y cuenta con recursos económicos limitados. Se caracteriza por tratar cada caso individualmente y no generalizar, ya que se tiene claro que cada persona tiene sus particularidades, así tengan el mismo diagnóstico, buscando satisfacer las necesidades que cada paciente tenga. Para la fabricación de los productos de apoyo se desarrolló un protocolo que busca que el producto final cumpla con los criterios de usabilidad (Facilidad de aprendizaje, eficiencia, entre otros.

  3. Age and sex correlation of Chlamydia trachomatis infections evaluated by the culture technique and by an enzyme immunosorbent assay, IDEIA

    DEFF Research Database (Denmark)

    Østergaard, Lars; Lundemose, AG; Birkelund, Svend

    1990-01-01

    tested by the tissue-culture technique, and the results were confirmed by the IDEIA (Boots-Celltech) enzyme-linked immunosorbent assay kit (EIA) for detection of C. trachomatis. The original smear was used for both culture and EIA. The EIA test was evaluated to have a sensitivity of 90...

  4. The right environment for the immunoassay

    International Nuclear Information System (INIS)

    Emon, J.M. Van; Gerlach, C.L.

    1995-01-01

    For the US Environmental Protection Agency (EPA), the first in-house research effort began in 1987, when results of an early immunoassay field study verified the technology's potential for environmental applications. Looking at the fundamental features of immunochemical reactions from the clinical laboratories, analytical chemists realized the potential value of these methods for hazardous waste site characterization and pesticide monitoring. Immunoassays rely on the interaction between an antibody and a target analyte. For environmental purposes, enzyme immunoassays are generally used. After the target analyte binds to the antibody, an enzymatic reaction yields a colorimetric change. This change, read visually or by a spectrophotometer, indicates the concentration of the target analyte. Promising results with assays for compounds (such as paraquat and pentachlorophenol) and compound groups (such as total petroleum hydrocarbons and polychlorinated biphenyls) spurred interest among various entrepreneurs. The first target market for immunoassays was environmental engineers and field crews who needed quick answers on-site to determine the direction of further remediation efforts

  5. Critical factors for EIA implementation: literature review and research options.

    Science.gov (United States)

    Zhang, Jie; Kørnøv, Lone; Christensen, Per

    2013-01-15

    After decades of development, the gap between expectations of Environment Impact Assessments (EIA) and their practical performance remains significant. Research has been done to identify the critical factors for an effective implementation of EIA. However, this research, to a large extent, has not been cumulated and analysed comprehensively according to the stages of the EIA process. This paper contributes to the critical review of the literature on EIA implementation and effectiveness by cumulating mainly empirical findings in an implementation theoretical perspective. It focuses on the links between different critical factors and how they relate to different stages in the EIA and thus influence the decision making process. After reviewing 33 refereed journal articles published between 1999 and 2011, we identified 203 notions of critical factors. Of these, 102 related to different stages defined in our comprehensive EIA implementation model, and 101 were identified as general factors related to the whole EIA system. The number of notions of stage factors and general factors is thus about equal. An overlap between stage factors and general factors was found, which demonstrates that critical factors function differently in different cases. The function of the critical factors is complex and it is difficult to determine contingencies and causations. In the sources we examined, there is evidently an imbalance between in-depth empirical research and general knowledge, and the paper offers some suggestions for future research. Copyright © 2012 Elsevier Ltd. All rights reserved.

  6. Correlations between calcineurin phosphatase inhibition and cyclosporine metabolites concentrations in kidney transplant recipients: implications for immunoassays

    DEFF Research Database (Denmark)

    Karamperis, N; Koefoed-Nielsen, PB; Brahe, P

    2006-01-01

    by the enzyme multiplied immunoassay technique (EMIT) and by the polyclonal fluorescence polarization immunoassay (pFPIA). Calcineurin phosphatase activity was measured by its ability to dephosphorylate a previously phosphorylated 19-amino acid peptide. We found that calcineurin phosphatase inhibition...... by inhibiting the enzyme calcineurin phosphatase. Determination of the enzyme's activity is one of the most promising pharmacodynamic markers. It is unknown how calcineurin phosphatase inhibition correlates with various cyclosporine monitoring assays and what is the potential impact of metabolites...

  7. Towards the development of a radioenzyme-immunoassay

    International Nuclear Information System (INIS)

    Schuurs, A.H.W.M.; Waart, M. v. d.

    1976-01-01

    We have tried to develop a very sensitive enzyme-immunoassay. For this purpose, a very sensitive radiochemical enzyme assay was used. HCG was chosen as test model and AChE as labelling enzyme. The test appeared to be much more sensitive than the normal enzymeimmunoassay. And, in comparison with RIA, it was about as sensitive but less time-consuming, and it makes use, in principle, of stable reagents. (orig./GSE) [de

  8. Method of immunoassay

    International Nuclear Information System (INIS)

    Scarisbrick, J.; Gard, T.; Hall, C.C.

    1983-01-01

    The invention relates to a method of immunoassay for prolactin using monoclonal lgG antibodies. The method preferably comprises the use of two different monoclonal antibodies which bind respectively at different antigenic sites on the prolactin molecule. One antibody is labelled and the other is immobilised on a water-insoluble carrier material, whereby an immunochemical complex comprising labelled antibody, prolactin and immobilised antibody is formed. (author)

  9. Reforming EIA systems: A critical review of proposals in Brazil

    International Nuclear Information System (INIS)

    Fonseca, Alberto; Sánchez, Luis Enrique; Ribeiro, José Claudio Junqueira

    2017-01-01

    Environmental Impact Assessment (EIA) systems are under pressure in many countries, driven by a call for efficiency and streamlining. Such a phenomenon is particularly clear in Brazil, where, in the past few years, a number of influential associations put forward documents proposing significant changes to environmental licensing and impact assessment regulations. So far, there is no publicly available information about any initiative towards scrutinizing those proposals. The objective of this study was to critically review the merits and drawbacks of the changes proposed in those documents. The analysis triangulated content analysis, focus group and online survey data. The focus group included ten seasoned Brazilian EIA specialists; the survey, based on Likert-scale and open-ended questions, resulted in 322 valid responses from EIA professionals. Results show that the proposals generally agree that the current EIA system, while playing a key role in mitigating impacts and enhancing project design, needs many changes. Nonetheless, the proposals neither offered solutions to overcome political, technical and budget barriers, nor established a sense of priority of the most urgent issues. Findings from the focus group and the survey signaled that a number of proposed actions might face public outcry, and that those changes that do not depend on legislative action are more likely to be implementable. Previous studies about EIA reform focused mostly on the context of developed countries after changes had taken place. This study, while addressing the perspective of a large developing country in a “before-reform” stage, shows that capacity-building is a key requirement in EIA reform. - Highlights: • Brazil's EIA system is under strong pressure for change. • Findings corroborate ineffectiveness in current system. • There are tensions as to the best approaches to overcome problems. • Exact effects of proposals are uncertain. • Low institutional capacity can play

  10. Status of immunoassay as an analytical tool in environmental investigations

    International Nuclear Information System (INIS)

    Van Emon, J.M.

    2000-01-01

    Immunoassay methods were initially applied in clinical situations where their sensitivity and selectivity were utilized for diagnostic purposes. In the 1970s, pesticide chemists realized the potential benefits of immunoassay methods for compounds difficult to analyze by gas chromatography. This transition of the technology has extended to the analysis of soil, water, food and other matrices of environmental and human exposure significance particularly for compounds difficult to analyze by chromatographic methods. The utility of radioimmunoassays and enzyme immunoassays for environmental investigations was recognized in the 1980s by the U.S. Environmental Protection Agency (U.S. EPA) with the initiation of an immunoassay development programme. The U.S. Department of Agriculture (USDA) and the U.S. Food and Drug Administration (PDA) have investigated immunoassays for the detection of residues in food both from an inspection and a contamination prevention perspective. Environmental immunoassays are providing rapid screening information as well as quantitative information to fulfill rigorous data quality objectives for monitoring programmes

  11. Interference in immunoassay

    International Nuclear Information System (INIS)

    Chapman, R.S.

    1998-01-01

    Interfering factors are evident in both limited reagent (radioimmunoassay) and excess reagent (immunometric assay) technologies and should be suspected whenever there is a discrepancy between analytical results and clinical findings in the investigation of particular diseases. The overall effect of interference in immunoassay is analytical bias in result, either positive or negative of variable magnitude. The interference maybe caused by a wide spectrum of factors from poor sample collection and handling to physiological factors e.g. lipaemia, heparin treatment, binding protein abnormalities, autoimmunity and drug treatments. The range of interfering factors is extensive and difficult to discuss effectively in a short review

  12. Immunoassays in monitoring biotechnological drugs.

    Science.gov (United States)

    Gygax, D; Botta, L; Ehrat, M; Graf, P; Lefèvre, G; Oroszlan, P; Pfister, C

    1996-08-01

    For the evaluation and interpretation of pharmacokinetic data reliable quantitative determinations are a requirement that can only be met by well-characterized and fully validated analytical methods. To cope with these requirements a method is being established that is based on an integrated and automated fiber-optic biospecific interaction analysis system (FOBIA) for immunoassays. Performance characteristics of this system used in monitoring of recombinant hirudin (CGP 39 393) are presented. Recombinant hirudin is a highly potent and selective inhibitor of human thrombin. Owing to its size and charge, recombinant hirudin is mainly eliminated by glomerular filtration. But only a fraction of the hirudin dose seems to be reabsorbed at the proximal tubule by luminal endocytosis and hydrolyzed by lysosomal enzymes, leaving approximately 50% of the dose to be extracted in the urine. Thus, renal clearance of recombinant hirudin in the absence of renal insufficiency appears to depend primarily on the glomerular filtration rate. During a 3-month i.v. tolerability study in dogs, some of the dogs developed antibodies against recombinant hirudin. The hirudin-antibody complex accumulated in plasma and apparent hirudin plasma concentrations were therefore much higher than expected from single-dose kinetics. Hirudin captured by antibodies showed an extended half-life and the hirudin-antibody complex is still pharmacologically active, as demonstrated by the observed increase in thrombin time. In conclusion, only appropriate analytical methods allow adequate monitoring and pharmacokinetic characterization of biotechnology drugs in biological materials.

  13. Diagnostic effectiveness of immunoassays systems for hepatitis C virus in samples from multi-transfusion patients

    International Nuclear Information System (INIS)

    Rivero Jimenez, Rene A; Merlin Linares, Julio C; Blanco de Armas, Madelin; Navea Leyva, Leonor M

    2009-01-01

    Hepatitis C virus (CHV) blood-transmission is a health problem in Cuba and in the world. Some types of diagnostic immunoassays have been developed for the blood certification and in general have a high diagnostic sensitivity and specificity in healthy donors. However, its behavior in samples from multi-transfusion patients could by less effective. To assess the diagnostic effectiveness of the UMELISA HCV third generation Cuban immunoassay (TecnoSUMA, S.A. La Habana), Cuba) in samples from multi-transfusion patients, in parallel, 335 sera from patients were processed by UBI HCV EIA 4.0 (United Biomedical, EE.UU) and UMELISA HCV third generation, and the samples with incongruous results were verified by PCR COBAS AmpliScreen HCV Test, v2 system (Roche, EE.UU.) Comparing the UMELISA HCV third generation system with the UBI HCV EIA 4.0 it was achieved a Sd of 95,8% CI(95%): 92,5-99,15 and a Ed of 100% CI (95%): 99,7-100, with IY: 0,96 (0,93-0,99) with k: 0,0582 ID (95%): 0,9276-0,9888, p = 0,000. Both immunoassay systems were satisfactory for immunodiagnosis of multi-transfusion patients

  14. Appraising the value of independent EIA follow-up verifiers

    Energy Technology Data Exchange (ETDEWEB)

    Wessels, Jan-Albert, E-mail: janalbert.wessels@nwu.ac.za [School of Geo and Spatial Sciences, Department of Geography and Environmental Management, North-West University, C/O Hoffman and Borcherd Street, Potchefstroom, 2520 (South Africa); Retief, Francois, E-mail: francois.retief@nwu.ac.za [School of Geo and Spatial Sciences, Department of Geography and Environmental Management, North-West University, C/O Hoffman and Borcherd Street, Potchefstroom, 2520 (South Africa); Morrison-Saunders, Angus, E-mail: A.Morrison-Saunders@murdoch.edu.au [School of Geo and Spatial Sciences, Department of Geography and Environmental Management, North-West University, C/O Hoffman and Borcherd Street, Potchefstroom, 2520 (South Africa); Environmental Assessment, School of Environmental Science, Murdoch University, Australia. (Australia)

    2015-01-15

    Independent Environmental Impact Assessment (EIA) follow-up verifiers such as monitoring agencies, checkers, supervisors and control officers are active on various construction sites across the world. There are, however, differing views on the value that these verifiers add and very limited learning in EIA has been drawn from independent verifiers. This paper aims to appraise how and to what extent independent EIA follow-up verifiers add value in major construction projects in the developing country context of South Africa. A framework for appraising the role of independent verifiers was established and four South African case studies were examined through a mixture of site visits, project document analysis, and interviews. Appraisal results were documented in the performance areas of: planning, doing, checking, acting, public participating and integration with other programs. The results indicate that independent verifiers add most value to major construction projects when involved with screening EIA requirements of new projects, allocation of financial and human resources, checking legal compliance, influencing implementation, reporting conformance results, community and stakeholder engagement, integration with self-responsibility programs such as environmental management systems (EMS), and controlling records. It was apparent that verifiers could be more creatively utilized in pre-construction preparation, providing feedback of knowledge into assessment of new projects, giving input to the planning and design phase of projects, and performance evaluation. The study confirms the benefits of proponent and regulator follow-up, specifically in having independent verifiers that disclose information, facilitate discussion among stakeholders, are adaptable and proactive, aid in the integration of EIA with other programs, and instill trust in EIA enforcement by conformance evaluation. Overall, the study provides insight on how to harness the learning opportunities

  15. Appraising the value of independent EIA follow-up verifiers

    International Nuclear Information System (INIS)

    Wessels, Jan-Albert; Retief, Francois; Morrison-Saunders, Angus

    2015-01-01

    Independent Environmental Impact Assessment (EIA) follow-up verifiers such as monitoring agencies, checkers, supervisors and control officers are active on various construction sites across the world. There are, however, differing views on the value that these verifiers add and very limited learning in EIA has been drawn from independent verifiers. This paper aims to appraise how and to what extent independent EIA follow-up verifiers add value in major construction projects in the developing country context of South Africa. A framework for appraising the role of independent verifiers was established and four South African case studies were examined through a mixture of site visits, project document analysis, and interviews. Appraisal results were documented in the performance areas of: planning, doing, checking, acting, public participating and integration with other programs. The results indicate that independent verifiers add most value to major construction projects when involved with screening EIA requirements of new projects, allocation of financial and human resources, checking legal compliance, influencing implementation, reporting conformance results, community and stakeholder engagement, integration with self-responsibility programs such as environmental management systems (EMS), and controlling records. It was apparent that verifiers could be more creatively utilized in pre-construction preparation, providing feedback of knowledge into assessment of new projects, giving input to the planning and design phase of projects, and performance evaluation. The study confirms the benefits of proponent and regulator follow-up, specifically in having independent verifiers that disclose information, facilitate discussion among stakeholders, are adaptable and proactive, aid in the integration of EIA with other programs, and instill trust in EIA enforcement by conformance evaluation. Overall, the study provides insight on how to harness the learning opportunities

  16. Lateral Flow Immunoassay.

    Science.gov (United States)

    Ching, Kathryn H

    2015-01-01

    Lateral flow immunoassays (LFIAs) are a staple in the field of rapid diagnostics. These small handheld devices require no specialized training or equipment to operate, and generate a result within minutes of sample application. They are an ideal format for many types of home test kits, for emergency responders and for food manufacturers and producers looking for a quick evaluation of a given sample. LFIAs rely on high quality monoclonal antibodies that recognize the analyte of interest. As monoclonal antibody technology becomes more accessible to smaller laboratories, there has been increased interest in developing LFIA prototypes for potential commercial manufacture. In this chapter, the basics of designing and building an LFIA prototype are described.

  17. Results of the HPL determination using a new enzymatic immunoassay - comparison with a commercial radioimmunoassay

    International Nuclear Information System (INIS)

    Leis, D.; Schneider, B.; Keller, A.; Braun, S.

    1982-01-01

    An enzymatic immunoassay recently developed by the firm Organon, Oss, Netherlands, to determine HPL in the serum of pregnant women was compared to a widely used radioimmunoassay marketed by the firm Amersham-Buchler, Braunschweig. The values determined showed a good correlation. The accuracy of measurement was within the generally accepted range for immunological assays of this kind. The assay procedures are comparable with regard to the expenditure of time and work. A great advantage of the EIA is due to the absence of radioactive substances, which permits this test to be performed in any laboratory equipped with a centrifuge and photometer. (orig.) [de

  18. A multicenter evaluation of the Biotest legionella urinary antigen EIA

    DEFF Research Database (Denmark)

    Harrison, Timothy; Uldum, Søren; Alexiou-Daniel, Stella

    1998-01-01

    assay and the Biotest EIA. Each examined: a standard panel of 12 coded urine samples (distributed by Biotest); a panel of 10 coded urine samples provided as part of a European external quality assurance (EQA) scheme; urine samples from patients with proven legionnaires' disease (LD); urine samples from...

  19. Expanding Hydro-Geotechnical Considerations in Eia Studies: A ...

    African Journals Online (AJOL)

    MICHAEL

    analysis in order to determine the local groundwater flow direction, and how ... aquifer or not. The aim of this paper, therefore, is to present an integrated study on the hydro-geotechnical aspects, for better understanding of their importance in EIA studies. GEOLOGY OF ... subsidence and deposition arising from a succession.

  20. Environmental Impact Assessment (Eia) On Project Design: An ...

    African Journals Online (AJOL)

    PROF. O. E. OSUAGWU

    2013-09-01

    Sep 1, 2013 ... Environmental Impact Assessment (Eia) On Project Design: An Emperical Approach. Ogbonna Achimba Chibueze. 1,. Nwakanma Ifeanyi Cosmas. 2 * and Njoku. Peter-Paul Chukwuemezie. 3. 1. School of Computing And Engineering Sciences, Babcock University, Nigeria. ( Acogbona06@Yahoo.Com ...

  1. Threatened biodiversity, the nema eia regulations and cultivation of ...

    African Journals Online (AJOL)

    Until such listing, unresolved legal questions that inhibited the effective consideration of biodiversity in agricultural decision-making prior to the promulgation of the NEMA EIA regulations are likely to persist—to the detriment of a globally imperilled biodiversity. This contribution sets out to identify some of the key issues that ...

  2. EIA model documentation: Electricity market module - electricity fuel dispatch

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1997-01-01

    This report documents the National Energy Modeling System Electricity Fuel Dispatch Submodule (EFD), a submodule of the Electricity Market Module (EMM) as it was used for EIA`s Annual Energy Outlook 1997. It replaces previous documentation dated March 1994 and subsequent yearly update revisions. The report catalogues and describes the model assumptions, computational methodology, parameter estimation techniques, model source code, and forecast results generated through the synthesis and scenario development based on these components. This document serves four purposes. First, it is a reference document providing a detailed description of the model for reviewers and potential users of the EFD including energy experts at the Energy Information Administration (EIA), other Federal agencies, state energy agencies, private firms such as utilities and consulting firms, and non-profit groups such as consumer and environmental groups. Second, this report meets the legal requirement of the Energy Information Administration (EIA) to provide adequate documentation in support of its statistical and forecast reports. Third, it facilitates continuity in model development by providing documentation which details model enhancements that were undertaken for AE097 and since the previous documentation. Last, because the major use of the EFD is to develop forecasts, this documentation explains the calculations, major inputs and assumptions which were used to generate the AE097.

  3. A Method for EIA Scoping of Wave Energy Converters

    DEFF Research Database (Denmark)

    Margheritini, Lucia; Hansen, Anne Merrild; Frigaard, Peter

    2012-01-01

    and marine environment, lack of coordination from the competent Authorities regulating devices deployment and conflicts of maritime areas utilization. The EIA within the consents process is central in the realization of full scale devices and often is the meeting point for technology, politics and public...

  4. 31 CFR 26.3 - Availability of Environmental Impact Assessment Summaries (EIA Summaries) and Environmental...

    Science.gov (United States)

    2010-07-01

    ... 31 Money and Finance: Treasury 1 2010-07-01 2010-07-01 false Availability of Environmental Impact Assessment Summaries (EIA Summaries) and Environmental Impact Assessments (EIAs). 26.3 Section 26.3 Money and... DEVELOPMENT BANDS (MDBs) § 26.3 Availability of Environmental Impact Assessment Summaries (EIA Summaries) and...

  5. Immunoassay for thymopoietin

    International Nuclear Information System (INIS)

    Goldstein, G.

    1979-01-01

    The patent describes the development of a radio-immunoassay for thymopoietin in biological samples. The method of raising antibodies to this polypeptide hormone is described. This is achieved by injecting a host animal with an antigen consisting of thymopoietin covalently bonded by glutaraldehyde to a carrier protein such as bovine serum albumin and equine globulin. Different methods of radiolabelling thymopoietin with 125 I for use as the tracer antigen are described. The Bolton-Hunter procedure was preferred to the chloramine-T method since direct iodination of the tyrosyl moieties of thymopoietin resulted in some loss of immunoreactivity. Systems for separating the antigen-antibody complex and unbound antigen are compared. Binding-inhibition curves for unlabelled thymopoietin in the assay employing polyethylene glycol separation showed a sensitivity of 5 ng thymopoietin/ml. However, using the double antibody or dextran coated charcoal separation techniques, the sensitivity of thymopoietin was 0.1 ng/ml. Thus these latter two procedures are thus especially suitable for measuring thymopoietin levels in serum or plasma samples. The assay was shown to be specific for thymopoietin, no significant displacement being produced by control polypeptides. (U.K.)

  6. Cumulative effects in Swedish EIA practice - difficulties and obstacles

    International Nuclear Information System (INIS)

    Waernbaeck, Antoienette; Hilding-Rydevik, Tuija

    2009-01-01

    The importance of considering cumulative effects (CE) in the context of environmental assessment is manifested in the EU regulations. The demands on the contents of Environmental Impact Assessment (EIA) and Strategic Environmental Assessment (SEA) documents explicitly ask for CE to be described. In Swedish environmental assessment documents CE are rarely described or included. The aim of this paper is to look into the reasons behind this fact in the Swedish context. The paper describes and analyse how actors implementing the EIA and SEA legislation in Sweden perceive the current situation in relation to the legislative demands and the inclusion of cumulative effects. Through semi-structured interviews the following questions have been explored: Is the phenomenon of CE discussed and included in the EIA/SEA process? What do the actors include in and what is their knowledge of the term and concept of CE? Which difficulties and obstacles do these actors experience and what possibilities for inclusion of CE do they see in the EIA/SEA process? A large number of obstacles and hindrances emerged from the interviews conducted. It can be concluded from the analysis that the will to act does seem to exist. A lack of knowledge in respect of how to include cumulative effects and a lack of clear regulations concerning how this should be done seem to be perceived as the main obstacles. The knowledge of the term and the phenomenon is furthermore quite narrow and not all encompassing. They experience that there is a lack of procedures in place. They also seem to lack knowledge of methods in relation to how to actually work, in practice, with CE and how to include CE in the EIA/SEA process. It can be stated that the existence of this poor picture in relation to practice concerning CE in the context of impact assessment mirrors the existing and so far rather vague demands in respect of the inclusion and assessment of CE in Swedish EIA and SEA legislation, regulations, guidelines and

  7. Understanding the long-term influence of EIA on organisational learning and transformation

    International Nuclear Information System (INIS)

    Jones, Megan; Morrison-Saunders, Angus

    2017-01-01

    This research is an attempt to verify the notion postulated by Robert Bartlett and Lynton Caldwell that the full benefits of environmental impact assessment (EIA) would take decades to be realized. While EIA is intended to directly influence decision-making regarding new development proposals, the process is also expected to lead to organisational learning and transformation over time. Our aim was to examine the influence of EIA on a single Western Australian proponent with sustained experience in the process to understand how EIA is used within the organisation and to seek evidence of transformation of the organisation's purpose and mission. The research reviewed literature in order to identify key influences of EIA on organisations, along with semi-structured staff interviews and document analysis for the case study organisation. Ascertaining causality that involvement in EIA processes influences or effects organisational learning and transformation is a challenge in the face of other societal events. Document analysis and interviewee data indicates that the action-forcing nature of EIA did influence proponent behavior through the creation of internal processes seeking to ensure robust design of new projects that would satisfy environmental protection expectations, without the need to trigger formal EIA. Evidence of EIA values and thinking were apparent within internal documentation, including the evolving mission statement. Our research indicates that participation in the EIA process can positively influence organisational learning and transformation by guiding internal change for decision-making. - Highlights: • The long-term influence of EIA on a proponent organisation is investigated. • EIA promotes internal organisational learning and transformation. • Analysis of mission statements can indicate the influence of EIA on organisations. • Organisations aligned with EIA values can reduce the need to engage in formal EIA.

  8. EIA practice in India and its evaluation using SWOT analysis

    International Nuclear Information System (INIS)

    Paliwal, Ritu

    2006-01-01

    In India Environmental Impact Assessment (EIA) has been formally introduced in 1994. It relied on the institutional framework that has a strong supporting legislative, administrative and procedural set-up. Both central and state authorities together are sharing the responsibility of its development and management. A Strength, Weakness, Opportunity and Threat (SWOT) analysis taken up in this article has suggested that there are several issues that need to be readdressed. It highlights several constraints, ranging from improper screening and scoping guidelines to ineffective monitoring and post project evaluation. The opportunities are realised as increasing public awareness, initiatives of environmental groups and business community and forward thinking to integrate environmental consideration into plans and policies. Poor governance, rapid economic reforms, and favours to small-scale units are some of the foreseen threats to the system. This article concludes with some suggestions to improve EIA process in India

  9. EIA model documentation: Electricity market module - electricity fuel dispatch

    International Nuclear Information System (INIS)

    1997-01-01

    This report documents the National Energy Modeling System Electricity Fuel Dispatch Submodule (EFD), a submodule of the Electricity Market Module (EMM) as it was used for EIA's Annual Energy Outlook 1997. It replaces previous documentation dated March 1994 and subsequent yearly update revisions. The report catalogues and describes the model assumptions, computational methodology, parameter estimation techniques, model source code, and forecast results generated through the synthesis and scenario development based on these components. This document serves four purposes. First, it is a reference document providing a detailed description of the model for reviewers and potential users of the EFD including energy experts at the Energy Information Administration (EIA), other Federal agencies, state energy agencies, private firms such as utilities and consulting firms, and non-profit groups such as consumer and environmental groups. Second, this report meets the legal requirement of the Energy Information Administration (EIA) to provide adequate documentation in support of its statistical and forecast reports. Third, it facilitates continuity in model development by providing documentation which details model enhancements that were undertaken for AE097 and since the previous documentation. Last, because the major use of the EFD is to develop forecasts, this documentation explains the calculations, major inputs and assumptions which were used to generate the AE097

  10. Development of a monoclonal antibody to urinary degradation products from the C-terminal telopeptide alpha 1 chain of type I collagen. Application in an enzyme Immunoassay and comparison to CrossLaps(TM) ELISA

    DEFF Research Database (Denmark)

    C, Fledelius; I, Kolding; P, Quist

    1997-01-01

    A monoclonal antibody MAbA7 was raised against a synthetic peptide having a sequence (EKAHDGGR) specific for a part of the C-telopeptide alpha 1 chain of type I collagen. MAbA7 was labelled with horseradish peroxide and used in a competitive one-step enzyme-linked immunosorbent assay (ELISA) for ...

  11. Variability in telavancin cross-reactivity among vancomycin immunoassays.

    Science.gov (United States)

    McConeghy, Kevin W; Liao, Siyun; Clark, Douglas; Worboys, Philip; Barriere, Steven L; Rodvold, Keith A

    2014-12-01

    Telavancin is a semisynthetic lipoglycopeptide with a dual mechanism of action against Gram-positive pathogens. Two brief reports have suggested potential cross-reactivity of telavancin with the vancomycin particle-enhanced turbidometric immunoassay (PETIA). The purpose of this study was to evaluate several commercially available vancomycin immunoassays (fluorescence polarization [FPIA], enzyme-multiplied immunoassays [EMIT], PETIA, and chemiluminescent immunoassay [CMIA]) for cross-reactivity with telavancin. Seven sites were selected to analyze serum samples for vancomycin. Each site received a set of samples (n = 18) which combined drug-free serum with telavancin, 7-OH telavancin metabolite, or vancomycin. Immunoassays demonstrating potential cross-reactivity were further evaluated by sending a duplicate sample set to multiple laboratories. Cross-reactivity was defined as the percent theoretical concentration (reported concentration/theoretical concentration × 100). No cross-reactivity was seen with FPIA or EMIT. Within the theoretical concentration range of 5 to 120 μg/ml of telavancin, the Synchron PETIA system reported vancomycin concentrations ranging from 4.7 to 54.2 μg/ml compared to vancomycin concentrations from 1.1 to 5.6 μg/ml for the Vista PETIA system. The Architect CMIA system reported vancomycin concentrations in the range of 0.27 to 0.97 μg/ml, whereas Advia Centaur XP CMIA reported vancomycin concentrations between 1.6 and 31.6 μg/ml. The Architect CMIA immunoassay had the lowest percent cross-reactivity (0.8 to 5.4%), while the Synchron PETIA immunoassay demonstrated the highest percent cross-reactivity (45.2 to 53.8%). Telavancin samples measured by liquid chromatography-mass spectroscopy were within 93.9 to 122% of theoretical concentrations. Vancomycin concentrations were not measured in any 7-OH telavancin-spiked sample. Vancomycin concentrations measured by liquid chromatography-mass spectroscopy were within 57.2 to 113% of

  12. Does enhanced regulation improve EIA report quality? Lessons from South Africa

    International Nuclear Information System (INIS)

    Sandham, L.A.; Heerden, A.J. van; Jones, C.E.; Retief, F.P.; Morrison-Saunders, A.N.

    2013-01-01

    Recently, various EIA systems have been subjected to system review processes with a view to improve performance. Many of these reviews resulted in some form of legislative reform. The South African Environmental Impact Assessment (EIA) regulations were modified in 2006 with the express intent to improve EIA effectiveness. In order to evaluate to what extent the desired outcome was achieved, the quality of EIA reports produced under the 2006 regulations was investigated for comparative analysis with the preceding regime. A sample of EIA reports from the two legislative regimes was reviewed using an adapted version of a well established method known colloquially as the “Lee and Colley” review package. Despite some improvements in certain aspects, overall report quality has decreased slightly from the 1997 EIA regime. It therefore appears that the modifications to the regulations, often heralded as the solution to improvements in performance have not resulted in improved quality of EIA reports. - Highlights: ► EIA regulations in South Africa were revised and became more comprehensive in 2006. ► The report quality of a sample of EIAs was reviewed using the Lee and Colley review package. ► Report quality showed a slight decline from the previous regulatory regime. ► EIA good practice needs flexibility rather than over-detailed regulation.

  13. Does enhanced regulation improve EIA report quality? Lessons from South Africa

    Energy Technology Data Exchange (ETDEWEB)

    Sandham, L.A., E-mail: luke.sandham@nwu.ac.za [Environmental Assessment Research Group, School of Geo and Spatial Sciences, North-West University, Private Bag X6001, Potchefstroom, 2520 (South Africa); Heerden, A.J. van [Environmental Assessment Research Group, School of Geo and Spatial Sciences, North-West University, Private Bag X6001, Potchefstroom, 2520 (South Africa); Jones, C.E. [School of Environment and Development, University of Manchester, Oxford Road, Manchester, M13 9PL (United Kingdom); Retief, F.P.; Morrison-Saunders, A.N. [Environmental Assessment Research Group, School of Geo and Spatial Sciences, North-West University, Private Bag X6001, Potchefstroom, 2520 (South Africa)

    2013-01-15

    Recently, various EIA systems have been subjected to system review processes with a view to improve performance. Many of these reviews resulted in some form of legislative reform. The South African Environmental Impact Assessment (EIA) regulations were modified in 2006 with the express intent to improve EIA effectiveness. In order to evaluate to what extent the desired outcome was achieved, the quality of EIA reports produced under the 2006 regulations was investigated for comparative analysis with the preceding regime. A sample of EIA reports from the two legislative regimes was reviewed using an adapted version of a well established method known colloquially as the 'Lee and Colley' review package. Despite some improvements in certain aspects, overall report quality has decreased slightly from the 1997 EIA regime. It therefore appears that the modifications to the regulations, often heralded as the solution to improvements in performance have not resulted in improved quality of EIA reports. - Highlights: Black-Right-Pointing-Pointer EIA regulations in South Africa were revised and became more comprehensive in 2006. Black-Right-Pointing-Pointer The report quality of a sample of EIAs was reviewed using the Lee and Colley review package. Black-Right-Pointing-Pointer Report quality showed a slight decline from the previous regulatory regime. Black-Right-Pointing-Pointer EIA good practice needs flexibility rather than over-detailed regulation.

  14. Molecularly Imprinted Polymer as an Antibody Substitution in Pseudo-immunoassays for Chemical Contaminants in Food and Environmental Samples.

    Science.gov (United States)

    Chen, Chaochao; Luo, Jiaxun; Li, Chenglong; Ma, Mingfang; Yu, Wenbo; Shen, Jianzhong; Wang, Zhanhui

    2018-03-21

    The chemical contaminants in food and the environment are quite harmful to food safety and human health. Rapid, accurate, and cheap detection can effectively control the potential risks derived from these chemical contaminants. Among all detection methods, the immunoassay based on the specific interaction of antibody-analyte is one of the most widely used techniques in the field. However, biological antibodies employed in the immunoassay usually cannot tolerate extreme conditions, resulting in an unstable state in both physical and chemical profiles. Molecularly imprinted polymers (MIPs) are a class of polymers with specific molecular recognition abilities, which are highly robust, showing excellent operational stability under a wide variety of conditions. Recently, MIPs have been used in biomimetic immunoassays for chemical contaminants as an antibody substitute in food and the environment. Here, we reviewed these applications of MIPs incorporated in different analytical platforms, such as enzyme-linked immunosorbent assay, fluorescent immunoassay, chemiluminescent immunoassay, electrochemical immunoassay, microfluidic paper-based immunoassay, and homogeneous immunoassay, and discussed current challenges and future trends in the use of MIPs in biomimetic immunoassays.

  15. Nanobody medicated immunoassay for ultrasensitive detection of cancer biomarker alpha-fetoprotein.

    Science.gov (United States)

    Chen, Jing; He, Qing-hua; Xu, Yang; Fu, Jin-heng; Li, Yan-ping; Tu, Zhui; Wang, Dan; Shu, Mei; Qiu, Yu-lou; Yang, Hong-wei; Liu, Yuan-yuan

    2016-01-15

    Immunoassay for cancer biomarkers plays an important role in cancer prevention and early diagnosis. To the development of immunoassay, the quality and stability of applied antibody is one of the key points to obtain reliability and high sensitivity for immunoassay. The main purpose of this study was to develop a novel immunoassay for ultrasensitive detection of cancer biomarker alpha-fetoprotein (AFP) based on nanobody against AFP. Two nanobodies which bind to AFP were selected from a phage display nanobody library by biopanning strategy. The prepared nanobodies are clonable, thermally stable and applied in both sandwich enzyme linked immunoassay (ELISA) and immuno-PCR assay for ultrasensitive detection of AFP. The limit detection of sandwich ELISA setup with optimized nanobodies was 0.48ng mL(-1), and the half of saturation concentration (SC50) value was 6.68±0.56ng mL(-1). These nanobodies were also used to develop an immuno-PCR assay for ultrasensitive detection of AFP, its limit detection values was 0.005ng mL(-1), and the linear range was 0.01-10,000ng mL(-1). These established immunoassays based on nanobodies were highly specific to AFP and with negligible cross reactivity with other tested caner biomarkers. Furthermore, this novel concept of nanobodies mediated immunoassay may provide potential applications in a general method for the ultrasensitive detection of various cancer biomarkers. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. EIA's Role in Energy Data Collection, With Some Notes on Water Data

    Science.gov (United States)

    Leckey, T. J.

    2017-12-01

    The U.S. Energy Information Administration (EIA) is the statistical and analytical agency within the U.S. Department of Energy. EIA collects, analyzes, and disseminates independent and impartial energy information to promote sound policymaking, efficient markets, and public understanding of energy and its interaction with the economy and the environment. EIA conducts a comprehensive data collection program that covers the full spectrum of energy sources, end uses, and energy flows. This presentation will describe EIA's authority to collect energy data, report on the range of energy areas currently collected by EIA, discuss some areas where energy information and water issues intersect, and describe the relatively few areas where EIA does collect a small amount of water data. The presentation will conclude with some thoughts about necessary components for effective collection of water data at the federal level.

  17. Uncertainty in Impact Assessment – EIA in Denmark

    DEFF Research Database (Denmark)

    Larsen, Sanne Vammen

    Uncertainty may be viewed as an inescapable part of the exercise of ex ante assessment of impacts of plans, programmes or projects – we do per definition not know the exact impacts before they unfold. Also, there is an increasing focus on integration of impacts such as climate change in impact...... as problematic, as this is important information for decision makers and public actors. Taking point of departure in these issues, this paper seeks to add to the discussions by presenting the results of a study on the handling of uncertainty in Environmental Impact Assessment (EIA) reports in Denmark. The study...

  18. The rationality of EIA forecasts under symmetric and asymmetric loss

    International Nuclear Information System (INIS)

    Auffhammer, Maximilian

    2007-01-01

    The United States Energy Information Administration publishes annual forecasts of nationally aggregated energy consumption, production, prices, intensity and GDP. These government issued forecasts often serve as reference cases in the calibration of simulation and econometric models, which climate and energy policy are based on. This study tests for rationality of published EIA forecasts under symmetric and asymmetric loss. We find strong empirical evidence of asymmetric loss for oil, coal and electricity prices as well as natural gas consumption, electricity sales, GDP and energy intensity. (author)

  19. Fabricating a UV-Vis and Raman Spectroscopy Immunoassay Platform.

    Science.gov (United States)

    Hanson, Cynthia; Israelsen, Nathan D; Sieverts, Michael; Vargis, Elizabeth

    2016-11-10

    Immunoassays are used to detect proteins based on the presence of associated antibodies. Because of their extensive use in research and clinical settings, a large infrastructure of immunoassay instruments and materials can be found. For example, 96- and 384-well polystyrene plates are available commercially and have a standard design to accommodate ultraviolet-visible (UV-Vis) spectroscopy machines from various manufacturers. In addition, a wide variety of immunoglobulins, detection tags, and blocking agents for customized immunoassay designs such as enzyme-linked immunosorbent assays (ELISA) are available. Despite the existing infrastructure, standard ELISA kits do not meet all research needs, requiring individualized immunoassay development, which can be expensive and time-consuming. For example, ELISA kits have low multiplexing (detection of more than one analyte at a time) capabilities as they usually depend on fluorescence or colorimetric methods for detection. Colorimetric and fluorescent-based analyses have limited multiplexing capabilities due to broad spectral peaks. In contrast, Raman spectroscopy-based methods have a much greater capability for multiplexing due to narrow emission peaks. Another advantage of Raman spectroscopy is that Raman reporters experience significantly less photobleaching than fluorescent tags 1 . Despite the advantages that Raman reporters have over fluorescent and colorimetric tags, protocols to fabricate Raman-based immunoassays are limited. The purpose of this paper is to provide a protocol to prepare functionalized probes to use in conjunction with polystyrene plates for direct detection of analytes by UV-Vis analysis and Raman spectroscopy. This protocol will allow researchers to take a do-it-yourself approach for future multi-analyte detection while capitalizing on pre-established infrastructure.

  20. Developing guidelines for economic evaluation of environmental impacts in EIAs. Part I

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2005-07-01

    Is the time right to introduce environmental evaluation into the Environmental Impact Assessment (EIA) system for large construction projects in China? The report analyses gaps to introducing environmental evaluation into EIAs and recommend how to bridge the gaps. The report also provides suggestions to the State Environmental Protection Administration on core elements of a guideline for environmental evaluation to include in the existing EIA guidelines. The report draws on international and Chinese research and best practice and conducts four case studies of environmental evaluation based on EIAs of investment projects e.g. a power plant, a waste water treatment plant, regional waste water irrigation, and a road construction project.

  1. Developing guidelines for economic evaluation of environmental impacts in EIAs. Part I

    International Nuclear Information System (INIS)

    2005-01-01

    Is the time right to introduce environmental evaluation into the Environmental Impact Assessment (EIA) system for large construction projects in China? The report analyses gaps to introducing environmental evaluation into EIAs and recommend how to bridge the gaps. The report also provides suggestions to the State Environmental Protection Administration on core elements of a guideline for environmental evaluation to include in the existing EIA guidelines. The report draws on international and Chinese research and best practice and conducts four case studies of environmental evaluation based on EIAs of investment projects e.g. a power plant, a waste water treatment plant, regional waste water irrigation, and a road construction project

  2. Europium nanoparticle-based high performing immunoassay for the screening of treponemal antibodies.

    Directory of Open Access Journals (Sweden)

    Sheikh M Talha

    Full Text Available Treponema pallidum subspecies pallidum (Tp is the causative agent of syphilis which mainly spreads through sexual contact, blood transfusion and perinatal route. In order to curtail the spread of the infection and to clinically manage the disease, timely, accurate and reliable diagnosis is very important. We have developed an immunoassay for the detection of treponemal antibodies in human serum or plasma samples. In vivo biotinylated and non-biotinylated versions of the recombinant antigen were designed by the fusion of three Tp-specific antigens namely Tp15, Tp17 and Tp47. These fusion antigens were expressed in E. coli and purified using single-step metal affinity chromatography. Biotinylated fusion antigen immobilized on streptavidin coated plate was used to capture the treponemal antibodies and the non-biotinylated antigen coated on europium nanoparticles was used as tracer. Assays with two different incubation times of 10 min and 1 h were developed, and following the incubation the europium fluorescence was measured using time-resolved fluorometry. The developed time-resolved fluorometric (TRF immunoassays were evaluated with in-house and commercial serum/plasma sample panels. For well-established treponemal antibodies positive or negative samples, the sensitivity of TRF immunoassay with 10 min incubation time was 97.4%, and of TRF immunoassay with 1 h incubation time was 98.7%, and the specificities of both the TRF immunoassays were 99.2%. For the samples with discordant results with the reference assays, both the TRF immunoassays showed better specificity than the Enzygnost syphilis enzyme immunoassay as a screening test. The two different incubation times did not have any significant effect on the signal to cutoff (S/Co ratios obtained with the two immunoassays (p=0.06. Our results indicate that the developed immunoassay with a short incubation time of 10 min has the potential to be used in clinical laboratories and in blood

  3. Immunoassay for determination of trilobolide

    Czech Academy of Sciences Publication Activity Database

    Huml, L.; Jurášek, M.; Mikšátková, P.; Zimmermann, T.; Tomanová, P.; Buděšínský, Miloš; Rottnerová, Z.; Šimková, M.; Harmatha, Juraj; Kmoníčková, Eva; Lapčík, O.; Drašar, P. B.

    2017-01-01

    Roč. 117, Jan (2017), s. 105-111 ISSN 0039-128X. [Conference on Isoprenoids /23./. Minsk, 04.09.2016-07.09.2016] Institutional support: RVO:61388963 ; RVO:68378041 Keywords : trilobolide * avidin-biotin * ELISA * Laser trilobum * synthesis * immunoassay Subject RIV: CE - Biochemistry; FR - Pharmacology ; Medidal Chemistry (UEM-P) OBOR OECD: Biochemical research methods; Pharmacology and pharmacy (UEM-P) Impact factor: 2.282, year: 2016

  4. The law concerning the environmental impact assessment. Vol. 1. Collection of regulations with an introduction to EIA law

    International Nuclear Information System (INIS)

    Peters, H.J.

    1995-01-01

    The present book contains all regulations relevant to EIA in compact form: The EU EIA Directive; the Federal Law on the EIA; the Procedural Rules of Atomic Energy Law; the Ninth Ordinance on the Federal Emissions Control Law including the pertinent general administrative regulation; the Federal Mining Law; the Federal Building Law; the Federal Regional Planning Law; and the EIA laws of the Laender such as implementing regulations, the Land EIA Laws, and the Land Planning Laws. There is a basic introduction to EIA law preceding this collection of regulations and laws. (orig./HP) [de

  5. 75 FR 43823 - Safety Zone; He'eia Kea Small Boat Harbor, Kaneohe Bay, Oahu, HI

    Science.gov (United States)

    2010-07-27

    ... DEPARTMENT OF HOMELAND SECURITY Coast Guard 33 CFR Part 165 [Docket No. USCG-2010-0458] RIN 1625-AA00 Safety Zone; He'eia Kea Small Boat Harbor, Kaneohe Bay, Oahu, HI AGENCY: Coast Guard, DHS. ACTION: Temporary Final Rule. SUMMARY: The Coast Guard is establishing a temporary safety zone in He'eia Kea Small...

  6. Newly released EIA-law. A palette of wishes?; Das neue UVP-Recht. Ein Wunschkonzert?

    Energy Technology Data Exchange (ETDEWEB)

    Feldmann, Ulrike

    2015-05-15

    The EU-directive on Environmental Impact Assessment (EIA) for certain public and private projects was once again amended in 2014 after controversial discussions, three previous modifications during 1985 and 2010 and an aggregation of all amendments within the EU-EIA-2011/92 directive. This newly released EU-EIA-directive 2014/52/EU (hereinafter RL 2014/52) is published within the EU-official journal EU L 124 p. 1 from 25.04.2014, came into force on 15. May 2014 and has to be adopted into international law until 16.05.2017. The modifications made are also valid, apart from certain exceptions, for licensing procedures (including decommissioning of nuclear power plants) in the field of nuclear energy, as far as they might have possible, significant environmental effects. The European EIA directive's 30th ''anniversary'' on 27.06.2015 raises the question, which substantial changes will come soon along with the newly released EU-EIA law. All in all it seems like if authorities and industry might get along with the newly released EU-EIA regulations. The responsible Federal Ministry for the Environment is already working on a first preliminary draft. It is under consideration if the EIA-regulation should be submitted additionally, beyond required amendments by EU-law, to a general revision. EIA-law remains exciting.

  7. Evaluation of United States-Licensed Human Immunodeficiency Virus Immunoassays for Detection of Group M Viral Variants

    OpenAIRE

    Koch, Walter H.; Sullivan, Patrick S.; Roberts, Charles; Francis, Kori; Downing, Robert; Mastro, Timothy D.; Nkengasong, John; Hu, Dale; Masciotra, Silvina; Schable, Charles; Lal, Renu B.

    2001-01-01

    Six Food and Drug Administration (FDA)-licensed human immunodeficiency virus type 1 (HIV-1) and HIV-1/2 immunoassays, including five enzyme immunoassays and one rapid test, were challenged with up to 250 serum samples collected from various global sites. The serum samples were from individuals known to be infected with variants of HIV-1 including group M subtypes A, B, B′, C, D, E, F, and G and group O. All immunoassays detected the vast majority of samples tested. Three samples produced low ...

  8. A review of EIA report quality in the North West province of South Africa

    International Nuclear Information System (INIS)

    Sandham, Luke A.; Pretorius, Hester M.

    2008-01-01

    The revised EIA regulations implemented on 3 July 2006 focused attention on the question of EIA effectiveness in South Africa. EIR quality review is one of the quality control functions contributing to EIA effectiveness within any EIA system, therefore the EIR quality review package developed by Lee and Colley was adapted and used to review the quality of a sample of 28 EIRs in the North West province of South Africa. Overall, 86% of the reports achieved satisfactory grades, with the descriptive and presentational elements of the EIRs more satisfactorily addressed, and the analytical components such as impact significance, addressed to a less satisfactory degree. EIR quality appears to be on par with international standards, but there are areas of distinct weakness. Further research is required to optimise quality review, and to reveal whether the new regulations have succeeded in addressing these weaknesses and made positive contributions to EIR quality, as a component of EIA effectiveness in South Africa

  9. EIA models and capacity building in Viet Nam: an analysis of development aid programs

    International Nuclear Information System (INIS)

    Doberstein, Brent

    2004-01-01

    There has been a decided lack of empirical research examining development aid agencies as 'agents of change' in environmental impact assessment (EIA) systems in developing countries, particularly research examining the model of environmental planning practice promoted by aid agencies as part of capacity building. This paper briefly traces a conceptual framework of EIA, then introduces the concept of 'EIA capacity building'. Using Viet Nam as a case study, the paper then outlines the empirical results of the research, focusing on the extent to which aid agency capacity-building programs promoted a Technical vs. Planning Model of EIA and on the coherence of capacity-building efforts across all aid programs. A discussion follows, where research results are interpreted within the Vietnamese context, and implications of research results are identified for three main groups of actors. The paper concludes by calling for development aid agencies to reconceptualise EIA capacity building as an opportunity to transform developing countries' development planning processes

  10. Environmental value assessment in a multidisciplinary EIA setting

    International Nuclear Information System (INIS)

    Erikstad, Lars; Lindblom, Inge; Jerpasen, Gro; Hanssen, Martin A.; Bekkby, Trine; Stabbetorp, Odd; Bakkestuen, Vegar

    2008-01-01

    Value assessment is a central element in an EIA for the understanding of the impacts of specified projects. The value assessment contains subjective elements and this may cause errors and difficulties in numeric value assessment methods. There is a need for transparent common criteria to promote discussion and understanding. A common criteria base already exists, but lack of communication between different management systems and different disciplines, all with different traditions in value assessment, makes the situation complex. In this article we have looked into the basic understanding of value linked to the investigation themes of natural environment, cultural heritage and society. The investigation themes linked to social science is difficult to incorporate into a common system, basically because they have less focus on land use and contain different value types. Much of the relevant literature about value assessment is linked to the assessment of sites of special interest as candidates for legal protection or conservation. In an EIA a much broader range of areas is introduced, including the 'every day landscape' with a lower and more general level of value. Together with a focus on mitigation and adjustments of plans, this results in a need for a more detailed value assessment scale than is normally in use today. We have suggested a new scale to ease communication between different disciplines and management systems. How we understand value is not constant over time, nor is the level of knowledge. This makes it necessary to sustain an ongoing debate on value assessment. The need for a dynamic value assessment system increases with the increasing use of database modelling, digital analysis of map data (GIS) etc. Lack of a ongoing value debate will rapidly lead to misleading and biased results

  11. Development of immunoassays for detecting clothianidin residue in agricultural products.

    Science.gov (United States)

    Li, Ming; Sheng, Enze; Cong, Lujing; Wang, Minghua

    2013-04-17

    Two enzyme-linked immunosorbent assays (ELISAs) based on polyclonal antibodies (PcAbs) for clothianidin are described: colorimetric detection format (ELISA) and pattern of chemiluminescent assay (CLEIA). Clothianidin hapten was synthesized and conjugated to bovine serum albumin (BSA) and ovalbumin (OVA) to produce immunogen and coating antigen. Anticlothianidin PcAbs were obtained from immunized New Zealand white rabbits. Under optimal conditions, the half-maximal inhibition concentration (IC₅₀) and the limit of detection (LOD, IC₂₀) of clothianidin were 0.046 and 0.0028 mg/L for the ELISA and 0.015 and 0.0014 mg/L for the CLEIA, respectively. There were no obvious cross-reactivities of the antibodies with its analogues except for dinotefuran. Recoveries of 76.4-116.4% for the immunoassays were achieved from spiked samples. The results of immunoassays for the spiked and authentic samples were largely consistent with gas chromatography. Therefore, the proposed immunoassays would be convenient and satisfactory analytical methods for the monitoring of clothianidin in agricultural products.

  12. Development of Polyclonal Antibody against Clenbuterol for Immunoassay Application

    Directory of Open Access Journals (Sweden)

    Nurul Ain A. Talib

    2018-03-01

    Full Text Available Development of an immunoassay for clenbuterol (CLB detection required an anti-CLB antibody as an important bioreceptor. In this study, we report our work on production and purification of a rabbit-derived polyclonal anti-CLB antibody. The antibody was then purified by nProtein A Sepharose affinity column and the antibody purity was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE analysis. The activities of purified antibody were evaluated based on high antibody titer determined from enzyme-linked immunosorbent assay (ELISA. The sensitivity and selectivity of this antibody was evaluated and exhibits negligible cross-reactivity to antibiotics other than β-agonist families. Evaluation of the antibody as bioreceptor in immunoassay was performed using direct competitive ELISA and exhibited linear calibration plot (R2 = 0.9484. The antibody was used to detect the content of CLB in spiked milk samples and the recovery of more than 92% indicating significant performance as bioreceptor for the development of a rapid and simple immunoassay.

  13. Competitive chemiluminescent anzyme immunoassay for vitamin B12 analysis in human milk.

    Science.gov (United States)

    Recent discoveries of matrix interferences by haptocorrin (HC) in human milk and serum show that past analyses of vitamin B12 in samples with high HC content might have been inaccurate (Lildballe et al., 2009; Carmel & Agrawal, 2012). We evaluated two competitive enzyme-binding immunoassays for seru...

  14. Good performance of an immunoassay based method for nevirapine measurements in human breast milk

    DEFF Research Database (Denmark)

    Salado-Rasmussen, Kirsten; Theilgaard, Zahra Persson; Chiduo, Mercy

    2011-01-01

    , requires complicated extraction techniques. The ARK method employs an immunoassay technology and requires a small sample volume (40 μL) and no pre-treatment of the samples. Methods: Commercial enzyme and antibody were used and calibration standards and quality controls were prepared from pooled breast milk...

  15. Alkaline phosphatase-fused repebody as a new format of immuno-reagent for an immunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Seo, Hyo-Deok; Lee, Joong-jae [Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon, 305-701 (Korea, Republic of); Kim, Yu Jung [Industrial Biotechnology and Bioenergy Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon (Korea, Republic of); Hantschel, Oliver [School of Life Sciences, École Polytechnique Fédérale de Lausanne (EPFL), Lausanne (Switzerland); Lee, Seung-Goo [Industrial Biotechnology and Bioenergy Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon (Korea, Republic of); Kim, Hak-Sung, E-mail: hskim76@kaist.ac.kr [Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon, 305-701 (Korea, Republic of)

    2017-01-15

    Enzyme-linked immunoassays based on an antibody-antigen interaction are widely used in biological and medical sciences. However, the conjugation of an enzyme to antibodies needs an additional chemical process, usually resulting in randomly cross-linked molecules and a loss of the binding affinity and enzyme activity. Herein, we present the development of an alkaline phosphatase-fused repebody as a new format of immuno-reagent for immunoassays. A repebody specifically binding to human TNF-α (hTNF-α) was selected through a phage display, and its binding affinity was increased up to 49 nM using a modular engineering approach. A monomeric alkaline phosphatase (mAP), which was previously isolated from a metagenome library, was genetically fused to the repebody as a signal generator, and the resulting repebody-mAP fusion protein was used for direct and sandwich immunoassays of hTNF-α. We demonstrate the utility and potential of the repebody-mAP fusion protein as an immuno-reagent by showing the sensitivity of 216 pg mL{sup −1} for hTNF-α in a sandwich immunoassay. Furthermore, this repebody-mAP fusion protein enabled the detection of hTNF-α spiked in a serum-supplemented medium with high accuracy and reproducibility. It is thus expected that a mAP-fused repebody can be broadly used as an immuno-reagent in immunoassays. - Highlights: • A human TNF-α (hTNF-α)-specific repebody was selected using a phage display. • A monomeric alkaline phosphatase (mAP) was genetically fused to the repebody. • mAP-fused repebody enabled detection of hTNF-α with high sensitivity and accuracy. • mAP-fused repebody can be widely used as a new immuno-reagent in immunoassays.

  16. Comparison of an enzyme-immunoassay with a radio-immunoassay ...

    African Journals Online (AJOL)

    1990-07-21

    Jul 21, 1990 ... The anti-HBs test uses a 'sandwich' method in which the patient's serum is added to beads coated with HBsAg and, after a period of incubation, HBsAg conjugated with iodine-. 125 which binds to any anti-HBs on the bead, is added. Department of Virology, University of the Witwatersrand and National ...

  17. Public participation in EIA in Hungary: Analysis through three case studies

    Energy Technology Data Exchange (ETDEWEB)

    Palerm, J.R. [Imperial College of Science, Technology and Medicine, London (United Kingdom)

    1999-03-01

    Public participation and environmental impact assessment (EIA) are recent developments in Hungary; in spite of this considerable advances have been made in their development. Hungarian EIA offers a range of public participation mechanisms depending on the year the permitting process began as well as the sector to which the project corresponds, offering a good range of examples to study and compare. Three case studies have been selected, each making use of different public participation schemes: (1) a hazardous waste incinerator, falling under the 1993 provisional EIA decree; (2) a power plant, falling under the 1993 provisional EIA decree as well as the 1994 Energy Act; and (3) a motorway previous to any EIA legislation but having to meet EBRD`s EIA requirements, the motorways planning process, and the developer`s own initiative for participation. The system`s strengths and weaknesses are identified, as well as lessons drawn from international EIA theory and practice, such as the need for including early public involvement and a formal scoping phase.

  18. Comparative assessment of EIA systems in MENA countries: challenges and prospects

    International Nuclear Information System (INIS)

    El-Fadl, Karma; El-Fadel, Mutasem

    2004-01-01

    Environmental impact assessment (EIA) was devised as a decision tool in response to the grand swell of ecocentric concerns to mediate between the technocentric view of continued development and the ability to create economic growth while overcoming environmental problems. The assimilation of the philosophy and practice of EIA into a broad range of cultures and political systems reflects the desire and need to integrate environmental considerations into the decision-making process. This paper unveils the status of EIA systems in countries of the Middle East and North Africa region through a comparative assessment of existing and planned legislation and procedures

  19. Enzyme immunoassay measurements of the molting hormone in ...

    African Journals Online (AJOL)

    Administrator

    2011-10-31

    Oct 31, 2011 ... C. pipiens and C. longiareolata (Diptera, Culicidae) were obtained from a stock colony and kept as previously described by Rehimi and Soltani (1999). Pyrex storage jars (80 by 100 mm) containing. 150 ml of tap water were maintained at temperature of 25°C and a photoperiod of 14:10 (L:D). Larvae were ...

  20. Oligopeptide-based enzyme immunoassay for ovine lentivirus antibody detection.

    OpenAIRE

    Kwang, J; Torres, J V

    1994-01-01

    Ovine progressive pneumonia virus (OPPV) is a lentivirus which causes a progressive disease in sheep. Immunodominant epitopes have been identified in the envelope gp40 glycoprotein. Synthetic peptides representing these regions are able to detect the presence of OPPV antibodies in 96% of infected sheep.

  1. Environmental impact monitoring in the EIA process of South Australia

    International Nuclear Information System (INIS)

    Ahammed, A.K.M. Rafique; Nixon, Bronte Merrick

    2006-01-01

    This paper presents the results of a detailed study of environmental impact monitoring (EIM) in South Australia by first establishing a context for EIM in the form of a literature review, and by then conducting a detailed content and practice analysis on EIA documents and on actual monitoring in practice. EIM in practice for specific projects was found to not correspond with either EIM proposals in the corresponding environmental impact statement, or EIM recommendations in the government's assessment report. EIM programmes appeared to be carried out at the discretion of the proponent, and no projects were found to have environmental impact monitoring that directly corresponded to either proposals in the EIS or recommendations in the assessment report. Reasons for this lack of correlation have been postulated to be a combination of a serious lack of the following factors: clearly assigned monitoring responsibilities, public accountability, clearly outlined monitoring timeframes, clearly outlined aims and objectives, lack of human resources, and lack of any EIM legislation. It should be noted that the study, upon which this paper is predominantly based, was undertaken in 1998. This study looked at developments that were approved (and subsequently built) under the Planning Act 1982. This Act was later replaced by the South Australian Development Act 1993. Relevant information from this study that has been reproduced in this paper was accurate at that time

  2. Application of EIA/SEA system in land use planning: Experience from Serbia

    Directory of Open Access Journals (Sweden)

    Stojanović Božidar

    2005-01-01

    Full Text Available This paper discusses the experience and current status of EIA/SEA procedures and assessment methodologies in Serbia, aiming to propose strategies that can lead to effective integration of the SEA in spatial planning. Institutional and practical problems with regard to the regulations of EIA/SEA were considered. Experience from the past decade shows that implementation of EIA system in Serbia has not been effective as expected. New legislation on EIA and SEA is harmonized with corresponding EU Directives. First steps in the application of the SEA show that the main issues are screening, scooping and decision making. According to the research results, it is suggested that extra evaluation processes should be incorporated into current assessment procedures to improve their scientific validity and integrity.

  3. The importance of context in delivering effective EIA: Case studies from East Africa

    International Nuclear Information System (INIS)

    Marara, Madeleine; Okello, Nick; Kuhanwa, Zainab; Douven, Wim; Beevers, Lindsay; Leentvaar, Jan

    2011-01-01

    This paper reviews and compares the condition of the environmental impact assessment (EIA) system in three countries in the East Africa region: Kenya, Rwanda and Tanzania. The criteria used for the evaluation and the comparison of each system are based on the elements of the legal, administrative and procedural frameworks, as well as the context in which they operate. These criteria are adapted from the evaluation and quality control criteria derived from a number of literature sources. The study reveals that the EIA systems of Kenya and Tanzania are at a similar stage in their development. The two countries, the first to introduce the EIA concept into their jurisdiction in this part of Africa, therefore have more experience than Rwanda in the practice of environmental impact assessment, where the legislation and process requires more time to mature both from the governmental and societal perspective. The analysis of the administrative and procedural frameworks highlights the weakness in the autonomy of the competent authority, in all three countries. Finally a major finding of this study is that the contextual set up i.e. the socio-economic and political situation plays an important role in the performance of an EIA system. The context in developing countries is very different from developed countries where the EIA concept originates. Interpreting EIA conditions in countries like Kenya, Rwanda and Tanzania requires that the analysis for determining the effectiveness of their systems should be undertaken within a relevant framework, taking into account the specific requirements of those countries.

  4. System-Based Assessments—Improving the Confidence in the EIA Process

    Directory of Open Access Journals (Sweden)

    Jeff Wilson

    2017-12-01

    Full Text Available This viewpoint article examines Environmental Impact Assessment (EIA practices in developed and transitioning nations, identifies weaknesses, and proposes a new quantitative approach. The literature indicates that there exists little to no standardization in EIA practice, transitioning nations rely on weak scientific impact analyses, and the establishment of baseline conditions is generally missing. The more fundamental issue is that the “receptor”-based approach leads to a qualitative and subjective EIA, as it does not adequately integrate the full measure of the complexity of ecosystems, ongoing project risks, and cumulative impacts. We propose the application of a new framework that aims to ensure full life cycle assessment of impacts applicable to any EIA process, within any jurisdictional context. System-Based EIA (SBEIA is based on modeling to predict changes and rests on data analysis with a statistically rigorous approach to assess impacts. This global approach uses technologies and methodologies that are typically applied to characterize ecosystem structure and functioning, including remote sensing, modeling, and in situ monitoring. The aim of this approach is to provide a method that can produce quantifiable reproducible values of impact and risk and move EIA towards its substantive goal of sustainable development. The adoption of this approach would provide a better evaluation of economic costs and benefits for all stakeholders.

  5. Advances in immunoassay of anabolic steroids

    International Nuclear Information System (INIS)

    Hampl, R.; Putz, Z.; Bicikova, M.; Starka, L.

    1985-01-01

    A specific RIA for the main nortestosterone metabolites, norandrosterone and noretiocholanolone and a rapid, automatizable modification of testosterone RIA in urine are described. Additionally, an enzymeimmunoassay (EIA) variant for detection of 17α-methyltestosterone and related steroids is suggested. (Auth.)

  6. Integrating education, training and communication for public involvement in EIA

    International Nuclear Information System (INIS)

    Oprea, Irina; Oprea, Marcel; Guta, Cornelia; Guta, Vasilica

    2003-01-01

    We are going towards a globalized world, this involving the integration of every activity and every person. The public involvement in the development process is evident, taking into account that any objective will affect the people and the negative feedback could influence the result of the investment. Generally the public could be influenced by amplification of negative evaluated consequences, resulting psychosocial effects leading to illness or anxieties. This problem will be resolved by the public access to information provided by experts. A real-time interactive communication system is proposed as an open tool in order to facilitate decision-making by access to rapid and reliable information. The main task of the system is to collect, process, display and exchange the information relative to environmental impact assessment (EIA), to provide assistance, to receive specific opinions, being also proposed for public understanding of the field. The education and training integration will mitigate the barriers, which may inhibit the interaction and communication process. To increase learning will assure specialists-public interaction and a good information flow for knowledge exchange. The paper will outline key approaches in reaching agreement on the people educational process importance. The impact of development will be available to the public revealing the positive consequences, such as increased employment and income. An effective way to avoid negative reactions consists of the extensive consultation to identify the concerns and needs of the public, the access to suggestive and attractive programs for education and training. The system is developed as a modern information module, integrated into complex international management systems. It can be placed everywhere, everybody could access the facilities for education, world experience and training. Providing a real-time response to citizen concerns, the system represents an economic and rapid way to mitigate the

  7. [Fluorescence polarization immunoassay of ractopamine].

    Science.gov (United States)

    Zvereva, E A; Shpakova, N A; Zherdev, A V; Kiu, L; Xu, C; Eremin, S A; Dzantiev, B B

    2016-01-01

    A technique was developed for fluorescence polarization immunoassay (FPIA) of ractopamine, a toxic low molecular weight nonsteroidal growth regulator belonging to the most controlled contaminants of food products of animal origin. The assay is based on the competition between a sample containing ractopamine and ractopamine–fluorophore conjugate for binding to antibodies. The competition is monitored via changes in the degree of fluorescence polarization for plane-polarized excitation light, which differs for the free and antibody-bound forms of the conjugate. The optimal assay conditions were established, ensuring a high accuracy and minimal detection limit. The developed assay demonstrated a detection limit of 1 ng/mL and a range of detectable concentrations of 2.3–50 ng/mL, which met the requirements of sanitary control. The duration of the analysis was 10 min. The possible application of the developed FPIA was demonstrated with testing of turkey meat. The speed and simplicity of the proposed assay define its efficiency as a screening tool for safety of foods.

  8. Variance function estimation for immunoassays

    International Nuclear Information System (INIS)

    Raab, G.M.; Thompson, R.; McKenzie, I.

    1980-01-01

    A computer program is described which implements a recently described, modified likelihood method of determining an appropriate weighting function to use when fitting immunoassay dose-response curves. The relationship between the variance of the response and its mean value is assumed to have an exponential form, and the best fit to this model is determined from the within-set variability of many small sets of repeated measurements. The program estimates the parameter of the exponential function with its estimated standard error, and tests the fit of the experimental data to the proposed model. Output options include a list of the actual and fitted standard deviation of the set of responses, a plot of actual and fitted standard deviation against the mean response, and an ordered list of the 10 sets of data with the largest ratios of actual to fitted standard deviation. The program has been designed for a laboratory user without computing or statistical expertise. The test-of-fit has proved valuable for identifying outlying responses, which may be excluded from further analysis by being set to negative values in the input file. (Auth.)

  9. Multiplex detection of plant pathogens using a microsphere immunoassay technology.

    Directory of Open Access Journals (Sweden)

    Ratthaphol Charlermroj

    Full Text Available Plant pathogens are a serious problem for seed export, plant disease control and plant quarantine. Rapid and accurate screening tests are urgently required to protect and prevent plant diseases spreading worldwide. A novel multiplex detection method was developed based on microsphere immunoassays to simultaneously detect four important plant pathogens: a fruit blotch bacterium Acidovorax avenae subsp. citrulli (Aac, chilli vein-banding mottle virus (CVbMV, potyvirus, watermelon silver mottle virus (WSMoV, tospovirus serogroup IV and melon yellow spot virus (MYSV, tospovirus. An antibody for each plant pathogen was linked on a fluorescence-coded magnetic microsphere set which was used to capture corresponding pathogen. The presence of pathogens was detected by R-phycoerythrin (RPE-labeled antibodies specific to the pathogens. The assay conditions were optimized by identifying appropriate antibody pairs, blocking buffer, concentration of RPE-labeled antibodies and assay time. Once conditions were optimized, the assay was able to detect all four plant pathogens precisely and accurately with substantially higher sensitivity than enzyme-linked immunosorbent assay (ELISA when spiked in buffer and in healthy watermelon leaf extract. The assay time of the microsphere immunoassay (1 hour was much shorter than that of ELISA (4 hours. This system was also shown to be capable of detecting the pathogens in naturally infected plant samples and is a major advancement in plant pathogen detection.

  10. Multiplex detection of plant pathogens using a microsphere immunoassay technology.

    Science.gov (United States)

    Charlermroj, Ratthaphol; Himananto, Orawan; Seepiban, Channarong; Kumpoosiri, Mallika; Warin, Nuchnard; Oplatowska, Michalina; Gajanandana, Oraprapai; Grant, Irene R; Karoonuthaisiri, Nitsara; Elliott, Christopher T

    2013-01-01

    Plant pathogens are a serious problem for seed export, plant disease control and plant quarantine. Rapid and accurate screening tests are urgently required to protect and prevent plant diseases spreading worldwide. A novel multiplex detection method was developed based on microsphere immunoassays to simultaneously detect four important plant pathogens: a fruit blotch bacterium Acidovorax avenae subsp. citrulli (Aac), chilli vein-banding mottle virus (CVbMV, potyvirus), watermelon silver mottle virus (WSMoV, tospovirus serogroup IV) and melon yellow spot virus (MYSV, tospovirus). An antibody for each plant pathogen was linked on a fluorescence-coded magnetic microsphere set which was used to capture corresponding pathogen. The presence of pathogens was detected by R-phycoerythrin (RPE)-labeled antibodies specific to the pathogens. The assay conditions were optimized by identifying appropriate antibody pairs, blocking buffer, concentration of RPE-labeled antibodies and assay time. Once conditions were optimized, the assay was able to detect all four plant pathogens precisely and accurately with substantially higher sensitivity than enzyme-linked immunosorbent assay (ELISA) when spiked in buffer and in healthy watermelon leaf extract. The assay time of the microsphere immunoassay (1 hour) was much shorter than that of ELISA (4 hours). This system was also shown to be capable of detecting the pathogens in naturally infected plant samples and is a major advancement in plant pathogen detection.

  11. Determination of phospholipid transfer proteins in rat tissues by immunoassays

    International Nuclear Information System (INIS)

    Teerlink, T.

    1983-01-01

    Several quantitative immunoassays have been developed for two phospholipid transfer proteins from rat liver, i.e. the phosphatidylcholine transfer protein and the non-specific lipid transfer protein. The development of a double-antibody radioimmunoassay for the phosphatidylcholine transfer protein is described. The transfer protein was labelled with iodine-125 by the mild glucose oxidase-lactoperoxidase method. Although less than one tyrosine residue per molecule of transfer protein was labelled, only 20% of the labelled transfer protein was immunoprecipitable. This value could be increased to 80% by purifying the labelled protein by affinity chromatography on a column of anti-phosphatidylcholine transfer protein-IgG coupled to Sepharose 4B. The radioimmunoassay was used to determine the levels of phosphatidylcholine transfer protein in homogenates and 105 000 xg supernatants from various rat tissues as well as several Morris hepatomas. An enzyme immunoassay for the non-specific lipid transfer protein is also described. The antiserum that was raised especially by the author was cross-reactive with the non-specific lipid transfer protein present in 105 000 xg supernatants from human, mouse and bovine liver. The non-specific lipid transfer protein lost its immunoreactivity upon labelling with iodine-125 using different labelling techniques. Therefore, a regular radioimmunoassay could not be developed. The results of these different assays were compared. (Auth.)

  12. Progressing quality control in environmental impact assessment beyond legislative compliance: An evaluation of the IEMA EIA Quality Mark certification scheme

    International Nuclear Information System (INIS)

    Bond, Alan; Fischer, Thomas B; Fothergill, Josh

    2017-01-01

    The effectiveness of Environmental Impact Assessment (EIA) systems is contingent on a number of control mechanisms: procedural; judicial; evaluative; public and government agency; professional; and development aid agency. If we assume that procedural and judicial controls are guaranteed in developed EIA systems, then progressing effectiveness towards an acceptable level depends on improving the performance of other control mechanisms over time. These other control mechanisms are either absent, or are typically centrally controlled, requiring public finances; this we argue is an unpopular model in times of greater Government austerity. Here we evaluate a market-based mechanism for improving the performance of evaluative and professional control mechanisms, the UK Institute of Environmental Management and Assessments' EIA Quality Mark. We do this by defining dimensions of effectiveness for the purposes of our evaluation, and by identifying international examples of the approaches taken to delivering the other control measures to validate the approach taken in the EIA Quality Mark. We then evaluate the EIA Quality Mark, when used in combination with legal procedures and an active judiciary, against the effectiveness dimensions and use time-series analysis of registrant data to examine its ability to progress practice. We conclude that the EIA Quality Mark has merit as a model for a market-based mechanism, and may prove a more financially palatable approach for delivering effective EIA in mature systems in countries that lack centralised agency oversight. It may, therefore, be of particular interest to some Member States of the European Union for ensuring forthcoming certification requirements stemming from recent amendments to the EIA Directive. - Highlights: • Quality control mechanisms in EIA are identified. • Effectiveness of EIA is conceptualised for evaluation purposes. • The UK IEMA EIA Quality Mark is introduced as a market-based mechanism. • The EIA

  13. Effective use of environmental impact assessments (EIAs) for geothermal development projects

    International Nuclear Information System (INIS)

    Goff, S.J.

    2000-01-01

    Both the developed and developing nations of the world would like to move toward a position of sustainable development while paying attention to the restoration of natural resources, improving the environment, and improving the quality of life. The impacts of geothermal development projects are generally positive. It is important, however, that the environmental issues associated with development be addressed in a systematic fashion. Drafted early in the project planning stage, a well-prepared Environmental Impact Assessment (EIA) can significantly add to the quality of the overall project. An EIA customarily ends with the decision to proceed with the project. The environmental analysis process could be more effective if regular monitoring, detailed in the EIA, continues during project implementation. Geothermal development EIAs should be analytic rather than encyclopedic, emphasizing the impacts most closely associated with energy sector development. Air quality, water resources and quality, geologic factors, and socioeconomic issues will invariably be the most important factors. The purpose of an EIA should not be to generate paperwork, but to enable superb response. The EIA should be intended to help public officials make decisions that are based on an understanding of environmental consequences and take proper actions. The EIA process has been defined in different ways throughout the world. In fact, it appears that no two countries have defined it in exactly the same way. Going hand in hand with the different approaches to the process is the wide variety of formats available. It is recommended that the world geothermal community work towards the adoption of a standard. The Latin American Energy Organization (OLADE) and the Inter-American Development Bank (IDB)(OLADE, 1993) prepared a guide that presents a comprehensive discussion of the environmental impacts and suggested mitigation alternatives associated with geothermal development projects. The OLADE guide

  14. Variability in the detection of macro TSH in different immunoassay systems.

    Science.gov (United States)

    Hattori, Naoki; Ishihara, Takashi; Shimatsu, Akira

    2016-01-01

    Macro TSH is a large molecular-sized TSH that is mostly a complex of TSH and IgG. Patients with macro TSH have elevated serum TSH and normal free thyroxine levels, mimicking subclinical hypothyroidism. The aim of this study was to clarify the degree of cross-reactivity of macro TSH to different commercial immunoassay systems. Screening for macro TSH was done using a polyethylene glycol (PEG) method and confirmed with gel filtration chromatography in serum samples from 1901 patients with subclinical hypothyroidism. Interference due to human anti-mouse antibodies (HAMA) was examined using HAMA blockers. TSH was measured with an enzyme immunoassay for the analysis of macro TSH. Serum TSH values in patients with macro TSH were also determined with the widely used commercial immunoassay platforms Elecsys, Centaur and Architect, and the detectability of macro TSH was compared among them. Gel filtration chromatography was performed with 174 serum samples with PEG-precipitable TSH ratios >75%. Twenty serum samples were found to contain large molecular-sized TSH, five of which were due to interference by HAMA. The prevalence of macro TSH was eventually 0.79% (15/1901). Commercial immunoassay systems variably recognized macro TSH. The Architect TSH immunoassay platform was the least reactive to macro TSH, but still recognized it in 60% of macro TSH-containing serum samples. There were no commercial TSH immunoassay platforms that did not cross-react with macro TSH. Screening for macro TSH should be performed before hormone replacement therapy is initiated for subclinical hypothyroidism. © 2016 European Society of Endocrinology.

  15. Evaluation of the EIA system on the Island of Mauritius and development of an environmental monitoring plan framework

    International Nuclear Information System (INIS)

    Ramjeawon, T.; Beedassy, R.

    2004-01-01

    The Environment Protection Act (EPA) in Mauritius provides for the application of an EIA license in respect of undertakings listed in its first schedule. Following the promulgation of the Act in June 1993, the Department of Environment (DOE) is issuing an average of 125 EIA licenses yearly. In general, the review exercise of an environmental impact assessment (EIA) is terminated once the license has been granted. The aim of this project was to evaluate the EIA system in Mauritius and to identify its weaknesses and strengths. One of the main weaknesses, besides the lack of EIA audits, is the absence of EIA follow-up monitoring. It is necessary to distinguish between monitoring done for regulatory purposes (compliance monitoring) and environmental monitoring related to the EIA. With the growth of the tourism industry on the island, coastal development projects have the potential to cause significant environmental impacts . A sample of EIA reports pertaining to this sector was assessed for its quality and follow-up mechanisms. Proposals for the contents of EIA Prediction Audits, Environmental Monitoring Plans (EMP) and the format for an EMP report are made

  16. A polymer lab-on-a-chip for magnetic immunoassay with on-chip sampling and detection capabilities.

    Science.gov (United States)

    Do, Jaephil; Ahn, Chong H

    2008-04-01

    This paper presents a new polymer lab-on-a-chip for magnetic bead-based immunoassay with fully on-chip sampling and detection capabilities, which provides a smart platform of magnetic immunoassay-based lab-on-a-chip for point-of-care testing (POCT) toward biochemical hazardous agent detection, food inspection or clinical diagnostics. In this new approach, the polymer lab-on-a-chip for magnetic bead-based immunoassay consists of a magnetic bead-based separator, an interdigitated array (IDA) micro electrode, and a microfluidic system, which are fully incorporated into a lab-on-a-chip on cyclic olefin copolymer (COC). Since the polymer lab-on-a-chip was realized using low cost, high throughput polymer microfabrication techniques such as micro injection molding and hot embossing method, a disposable polymer lab-on-a-chip for the magnetic bead-based immunoassay can be successfully realized in a disposable platform. With this newly developed polymer lab-on-a-chip, an enzyme-labelled electrochemical immunoassay (ECIA) was performed using magnetic beads as the mobile solid support, and the final enzyme product produced from the ECIA was measured using chronoamperometry. A sampling and detection of as low as 16.4 ng mL(-1) of mouse IgG has been successfully performed in 35 min for the entire procedure.

  17. Environmental agreements, EIA follow-up and aboriginal participation in environmental management: The Canadian experience

    International Nuclear Information System (INIS)

    O'Faircheallaigh, Ciaran

    2007-01-01

    During the last decade a number of environmental agreements (EAs) have been negotiated in Canada involving industry, government and Aboriginal peoples. This article draws on the Canadian experience to consider the potential of such negotiated agreements to address two issues widely recognised in academic and policy debates on environmental impact assessment (EIA) and environmental management. The first relates to the need to secure indigenous participation in environmental management of major projects that affect indigenous peoples. The second and broader issue involves the necessity for specific initiatives to ensure effective follow-up of EIA. The Canadian experience indicates that negotiated environmental agreements have considerable potential to address both issues. However, if this potential is to be realized, greater effort must be made to develop structures and processes specifically designed to encourage Aboriginal participation; and EAs must themselves provide the financial and other resource required to support EIA follow-up and Aboriginal participation

  18. Development of an ultrasensitive immunoassay for detecting tartrazine.

    Science.gov (United States)

    Li, Zhuokun; Song, Shanshan; Xu, Liguang; Kuang, Hua; Guo, Shidong; Xu, Chuanlai

    2013-06-25

    We have developed an ultrasensitive indirect competitive enzyme-linked immunosorbent assay for the determination of tartrazine. Two carboxylated analogues of tartrazine with different spacer lengths, and one derivative from commercial tartrazine after a little chemical modification, were synthesized as haptens in order to produce antibodies specific to tartrazine. The effect of sulfonic acid groups on the hapten structure of tartrazine was also studied carefully for the first time. A most specific monoclonal antibody against tartrazine was created and exhibited an IC50 value of 0.105 ng/mL and a limit of detection of 0.014 ng/mL, with no cross-reactivity to other structurally-related pigments. The established immunoassay was applied to the determination of tartrazine in fortified samples of orange juice and in real positive samples of carbonated beverages.

  19. Development of an Ultrasensitive Immunoassay for Detecting Tartrazine

    Directory of Open Access Journals (Sweden)

    Chuanlai Xu

    2013-06-01

    Full Text Available We have developed an ultrasensitive indirect competitive enzyme-linked immunosorbent assay for the determination of tartrazine. Two carboxylated analogues of tartrazine with different spacer lengths, and one derivative from commercial tartrazine after a little chemical modification, were synthesized as haptens in order to produce antibodies specific to tartrazine. The effect of sulfonic acid groups on the hapten structure of tartrazine was also studied carefully for the first time. A most specific monoclonal antibody against tartrazine was created and exhibited an IC50 value of 0.105 ng/mL and a limit of detection of 0.014 ng/mL, with no cross-reactivity to other structurally-related pigments. The established immunoassay was applied to the determination of tartrazine in fortified samples of orange juice and in real positive samples of carbonated beverages.

  20. Consideration of climate change impacts and adaptation in EIA practice — Perspectives of actors in Austria and Germany

    International Nuclear Information System (INIS)

    Jiricka, Alexandra; Formayer, Herbert; Schmidt, Anna; Völler, Sonja; Leitner, Markus; Fischer, Thomas B.; Wachter, Thomas F.

    2016-01-01

    Current political discussions and developments indicate the importance and urgency of incorporating climate change considerations into EIA processes. The recent revision of the EU Directive 2014/52/EU on Environmental Impact Assessment (EIA) requires changes in the EIA practice of the EU member states. This paper investigates the extent to which the Environmental Impact Assessment (EIA) can contribute to an early consideration of climate change consequences in planning processes. In particular the roles of different actors in order to incorporate climate change impacts and adaptation into project planning subject to EIA at the appropriate levels are a core topic. Semi-structured expert interviews were carried out with representatives of the main infrastructure companies and institutions responsible in these sectors in Austria, which have to carry out EIA regularly. In a second step expert interviews were conducted with EIA assessors and EIA authorities in Austria and Germany, in order to examine the extent to which climate-based changes are already considered in EIA processes. This paper aims to discuss the different perspectives in the current EIA practice with regard to integrating climate change impacts as well as barriers and solutions identified by the groups of actors involved, namely project developers, environmental competent authorities and consultants (EIA assessors/practitioners). The interviews show that different groups of actors consider the topic to different degrees. Downscaling of climate change scenarios is in this context both, a critical issue with regards to availability of data and costs. Furthermore, assistance for the interpretation of relevant impacts, to be deducted from climate change scenarios, on the specific environmental issues in the area is needed. The main barriers identified by the EIA experts therefore include a lack of data as well as general uncertainty as to how far climate change should be considered in the process without

  1. Consideration of climate change impacts and adaptation in EIA practice — Perspectives of actors in Austria and Germany

    Energy Technology Data Exchange (ETDEWEB)

    Jiricka, Alexandra, E-mail: alexandra.jiricka@boku.ac.at [University of Natural Resources and Applied Life Sciences, Vienna (Austria); Department of Landscape, Spatial and Infrastructure Sciences, Institute for Landscape Development, Recreation and Conservation Planning, Peter-Jordan-Straße 82, 1190 Vienna (Austria); Formayer, Herbert; Schmidt, Anna [University of Natural Resources and Applied Life Sciences, Vienna (Austria); Department of Landscape, Spatial and Infrastructure Sciences, Institute for Landscape Development, Recreation and Conservation Planning, Peter-Jordan-Straße 82, 1190 Vienna (Austria); Völler, Sonja; Leitner, Markus [Environment Agency Austria, Environmental Impact Assessment and Climate Change, Spittelauer Lände 5, 1090 Wien (Austria); Fischer, Thomas B. [Environmental Assessment and Management, School of Environmental Sciences, University of Liverpool, 74 Bedford Street South, Liverpool L69 7ZQ (United Kingdom); Wachter, Thomas F. [Büro für Umweltplanung Dr. Wachter, Wiesnerring 2c, Hamburg (Germany)

    2016-02-15

    Current political discussions and developments indicate the importance and urgency of incorporating climate change considerations into EIA processes. The recent revision of the EU Directive 2014/52/EU on Environmental Impact Assessment (EIA) requires changes in the EIA practice of the EU member states. This paper investigates the extent to which the Environmental Impact Assessment (EIA) can contribute to an early consideration of climate change consequences in planning processes. In particular the roles of different actors in order to incorporate climate change impacts and adaptation into project planning subject to EIA at the appropriate levels are a core topic. Semi-structured expert interviews were carried out with representatives of the main infrastructure companies and institutions responsible in these sectors in Austria, which have to carry out EIA regularly. In a second step expert interviews were conducted with EIA assessors and EIA authorities in Austria and Germany, in order to examine the extent to which climate-based changes are already considered in EIA processes. This paper aims to discuss the different perspectives in the current EIA practice with regard to integrating climate change impacts as well as barriers and solutions identified by the groups of actors involved, namely project developers, environmental competent authorities and consultants (EIA assessors/practitioners). The interviews show that different groups of actors consider the topic to different degrees. Downscaling of climate change scenarios is in this context both, a critical issue with regards to availability of data and costs. Furthermore, assistance for the interpretation of relevant impacts, to be deducted from climate change scenarios, on the specific environmental issues in the area is needed. The main barriers identified by the EIA experts therefore include a lack of data as well as general uncertainty as to how far climate change should be considered in the process without

  2. Simultaneous determination of chloramphenicol, florfenicol and florfenicol amine in ham sausage with a hybrid chemiluminescent immunoassay.

    Science.gov (United States)

    Tao, Xiaoqi; Jiang, Haiyang; Yu, Xuezhi; Zhu, Jinghui; Wang, Xia; Wang, Zhanhui; Niu, Lanlan; Wu, Xiaoping; Shen, Jianzhong

    2013-01-01

    A novel chemiluminescent immunoassay utilising two types of primary antibodies (murine monoclonal antibody and rabbit polyclonal antibody) and two types of horseradish peroxidase-labelled secondary antibodies was established for simultaneously detecting multiple amphenicol residues in ham sausage. After combining the extract procedure of the target amphenicol into one simplified method, this hybrid chemiluminescent immunoassay could screen chloramphenicol (CAP), florfenicol (FF) and its metabolite florfenicol amine (FFA) at the same time by adding the corresponding secondary antibody. Ham sausage samples were analysed by using this hybrid immunoassay, with LODs of CAP being 0.01 μg kg⁻¹, of FF being 2.8 μg kg⁻¹ and of FFA being 3.0 μg kg⁻¹. The applicability of the proposed method has been validated by determining CAP, FF and FFA in ham sausage samples with satisfactory results. Good recoveries and high correlation with traditional enzyme-linked immunosorbent assay and LC-MS/MS results illustrated that the developed hybrid chemiluminescent immunoassay could screen high-throughput ultra-trace amphenicol residues effectively at one time.

  3. Mandated monitoring of post-project impacts in the Czech EIA

    International Nuclear Information System (INIS)

    Branis, Martin; Christopoulos, Stamatios

    2005-01-01

    Altogether, 801 documents of consent (50% of all) issued under the EIA Act No. 244/1992 by the competent authorities in the Czech Republic between 1993 and 2001 were studied. The aim of the analysis was to find and characterize conditions prescribing to the developer to perform ex-ante and ex-post monitoring of potential impacts of projects submitted for approval. It was found that each of the studied documents (standpoints) contained on average three to four conditions prescribing to collect data on various environmental factors during the preparation, implementation and/or operation phase of the development in question. The number of monitoring conditions contained in the standpoints issued by the Ministry of Environment as well as by the District Offices increased during the period studied from about two to five per project indicating a growing interest in and/or need to obtaining such data. Even though there is a good legal background for collecting monitoring data from implementation and operation phase of new developments, the Czech EIA Act (similarly as EIA acts in other countries) does not provide any practical background for this activity. Without relevant institutional, personal and financial support the possibility to impose post-project monitoring to the developer remains rather a challenge to, not advantage of the Czech EIA Act

  4. Screening for EIA in India: enhancing effectiveness through ecological carrying capacity approach.

    Science.gov (United States)

    Rajaram, T; Das, Ashutosh

    2011-01-01

    Developing countries across the world have embraced the policy of high economic growth as a means to reduce poverty. This economic growth largely based on industrial output is fast degrading the ecosystems, jeopardizing their long term sustainability. Environmental Impact Assessment (EIA) has long been recognized as a tool which can help in protecting the ecosystems and aid sustainable development. The Screening guidelines for EIA reflect the level of commitment the nation displays towards tightening its environmental protection system. The paper analyses the screening process for EIA in India and dissects the rationale behind the exclusions and thresholds set in the screening process. The screening process in India is compared with that of the European Union with the aim of understanding the extent of deviations from a screening approach in the context of better economic development. It is found that the Indian system excludes many activities from the purview of screening itself when compared to the EU. The constraints responsible for these exclusions are discussed and the shortcomings of the current command and control system of environmental management in India are also explained. It is suggested that an ecosystem carrying capacity based management system can provide significant inputs to enhance the effectiveness of EIA process from screening to monitoring. Copyright © 2010 Elsevier Ltd. All rights reserved.

  5. An experiential approach to improving the integration of knowledge during EIA in transport planning

    Energy Technology Data Exchange (ETDEWEB)

    Soria-Lara, Julio A., E-mail: julio.soria-lara@ouce.ox.ac.uk [Transport Research Unit, School of Geography and the Environment, University of Oxford, South Parks Road, Oxford OX1 3QY (United Kingdom); Bertolini, Luca, E-mail: l.bertolini@uva.nl [Centre for Urban Studies, University of Amsterdam, Nieuwe Achtergracht 166 1018 WV Amsterdam (Netherlands); Brömmelstroet, Marco te, E-mail: M.C.G.teBrommelstroet@uva.nl [Centre for Urban Studies, University of Amsterdam, Nieuwe Achtergracht 166 1018 WV Amsterdam (Netherlands)

    2016-01-15

    The integration of knowledge from stakeholders and the public at large is seen as one of the biggest process-related barriers during the scoping phase of EIA application in transport planning. While the academic literature offers abundant analyses, discussions and suggestions how to overcome this problem, the proposed solutions are yet to be adequately tested in practice. In order to address this gap, we test the effectiveness of a set of interventions and trigger mechanisms for improving different aspects of knowledge integration. The interventions are tested in an experiential study with two sequential cases, representing “close-to-real-life” conditions, in the context of two cities in Andalusia, Spain. In general terms, the participants perceived that the integration of knowledge improved during the simulation of the EIA scoping phase. Certain shortcomings were also discussed, fundamentally related to how the time spent during the scoping phase was crucial to lead an effective learning process between the involved people. The study concludes with a reflection on the effectiveness of the tested interventions according to similarities and differences obtained from the two experiential case studies, as well as with a discussion of the potential to generate new knowledge through the use of experiential studies in EIA practice. - Highlights: • It tests a set of interventions and mechanisms to improve the integration of knowledge. • The scoping phase of EIA is simulated to assess the effectiveness of interventions. • Two sequential case studies are used.

  6. The fabrication of magnetic particle-based chemiluminescence immunoassay for human epididymis protein-4 detection in ovarian cancer

    Directory of Open Access Journals (Sweden)

    Xiaoling Fu

    2018-03-01

    Full Text Available The magnetic particles have a significant influence on the immunoassay detection and cancer therapy. Herein, the chemiluminescence immunoassay combined with the magnetic particles (MPCLIA was presented for the clinical determination and analysis of human epididymis protein 4 (HE4 in the human serum. Under the optimized experiment conditions, the secure MPCLIA method can detect HE4 in the broader range of 0–1000 pmol/L, with a lower detection limit of 1.35 pmol/L. The satisfactory recovery rate of the method in the serum ranged from 83.62% to 105.10%, which was well within the requirement of clinical analysis. Moreover, the results showed the good correlation with enzyme-linked immunosorbent assay (ELISA, with the correlation coefficient of 0.9589. This proposed method has been successfully applied to the clinical determination of HE4 in the human serum. Keywords: Chemiluminescence immunoassay, Magnetic particles, Human epididymis protein 4

  7. Diagnóstico de leptospirosis: evaluación de un enzimoinmunoensayo en fase sólida en diferentes etapas de la enfermedad Diagnosis of leptospirosis: evaluation of a solid-phase enzyme immunoassay in different stages of the disease

    Directory of Open Access Journals (Sweden)

    Norma B. Vanasco

    2007-06-01

    solid-phase enzyme immunoassay (ELISA for genus-specific immunoglobulin G (IgG determination with leptospirosis and to evaluate the ELISA in different stages of the disease. METHODS: A total of 1 077 serum samples from 812 patients with suspected leptospirosis were analyzed. The samples had come from diagnoses done in the laboratory of the National Institute of Respiratory Diseases (Instituto Nacional de Enfermedades Respiratorias, in the city of Santa Fe, Argentina, between 1999 and 2005. Included in the study were 182 confirmed cases (267 samples, 167 negative cases (293 samples, and 40 probable cases (60 samples (based on case definitions based on the results from the microscopic agglutination test (MAT, leukocyte counts, and neutrophilia values. Each sample was classified, according to the days of the natural history of disease, into one of three stages: first ( 25 days. The antigen used in the ELISA was an extract of a mixture of pyrogenes and tarassovi serovars cultivated in a liquid medium, treated with ultrasound, and immobilized by adsorption on polystyrene plates. As a secondary antibody, a peroxidase-conjugated goat anti-human IgG monoclonal antibody was used. The cutoff value, sensitivity, and specificity of the ELISA were determined using the definitions of confirmed cases and of negatives cases as the standard. In order to determine the optimal cutoff value, the area under the receiver operating characteristic curve was calculated. RESULTS: The sensitivity of the evaluated test was much higher in the second stage (93.2% than in either the first stage (68.1% or the third stage (78.8%. The specificity increased gradually from 96.3% in the first stage to 100% in the third stage. CONCLUSIONS: Our results indicate that this ELISA test can be a very useful complement to the MAT for the diagnosis of leptospirosis in all the stages and, in particular, in order to diagnose acute disease sooner.

  8. Determination of triiodothyronine in serum by enzyme- and radioimmunoassay

    International Nuclear Information System (INIS)

    Oellerich, M.; Haindl, H.; Medizinische Hochschule Hannover

    1981-01-01

    An evaluation of a heterogeneous enzyme immunoassay for determination of triiodothyronine in serum (Enzymun-Test T 3 , Boehringer Mannheim) is presented. The enzyme immunoassay was compared with the laboratory routine radioimmunoassay. The precision of both assays was satisfactory at triiodothyronine concentrations between 1.0 and 8.0 nmol/l (coefficients of variation from day to day 3 from 96-104% and with the radioimmunoassay from 88-111%. A comparison of the results obtained by Enzymun-Test T 3 and the radioimmunoassay in a series of 103 patients showed a good correlation between both methods. L-thyroxine did not cause a relevant cross-reaction in the enzyme immunoassay. About 20 unknown samples can be analyzed in triplicate by Enzymun-Test T 3 within 260 minutes. (orig.) [de

  9. An analysis framework for characterizing and explaining development of EIA legislation in developing countries—Illustrated for Georgia, Ghana and Yemen

    International Nuclear Information System (INIS)

    Kolhoff, Arend J.; Driessen, Peter P.J.; Runhaar, Hens A.C.

    2013-01-01

    Actors in the field of international development co-operation supporting the development of EIA legislation in developing countries often do not achieve the results envisaged. The performance of EIA in these countries often remains weak. One reason, we assume, is that often those actors support the establishment of overly ambitious EIA legislation that cannot achieve its objectives in the light of constraining contexts. To provide more effective support we need to better understand the enabling and constraining contextual factors that influence the development of EIA legislation and to which support actors should align itself. In this article a new analysis framework for classifying, characterizing and explaining the development of EIA legislation is described, measured in terms of ambition levels. Ambitions are defined as intentions the EIA authorities aim to fulfill, expressed in formal EIA legislation. Three country cases, Yemen, Georgia and Ghana are used to illustrate the usefulness of our framework and as a first test to refine the framework. We have formulated the following five hypotheses that complement and refine our analysis framework. One, EIA legislation may develop multilinearly in terms of ambition levels. Two, ambitions in EIA legislation seem to be influenced to a great extent by the power and capacity of, on the one hand, the environmental authorities supporting EIA and, on the other hand, the sector authorities hindering the development of EIA. Three, the political system is the most important context factor influencing the rules of policy-making and the power of the different actors involved. Four, the importance of context factors on the development of ambitions is dependent on the phase of EIA system development. Five, some ambitions seem to be influenced by particular factors; for instance the ambitions for the object of study seem to be influenced by the level of environmental awareness of the sector ministries and parliament. The analysis

  10. Development of national immunoassay reagent programmes

    International Nuclear Information System (INIS)

    Sufi, S.B.; Micallef, J.V.; Ahsan, R.; Goncharov, N.P.

    1992-01-01

    Despite the existence of networks of fully equipped laboratories with well-trained staff, the availability of immunodiagnostic services in developing countries is often limited by the high cost of imported kits. There are a number of ways of tackling this problem, ranging from bulk purchase of kits or reagents to local development and production of assay systems. Argentina/Chile, China, Cuba/Mexico, and Thailand are amongst the countries which have established local immunoassay reagent programmes to manufacture low cost, high quality immunoassay reagents. Kits from these projects are now beginning to become available, and it is hoped that they will promote national diagnostic services and research, as well as stimulating the development of reagent programmes for other analytes. (author). 4 refs, 1 tab

  11. A quantitative method to analyze the quality of EIA information in wind energy development and avian/bat assessments

    International Nuclear Information System (INIS)

    Chang, Tony; Nielsen, Erik; Auberle, William; Solop, Frederic I.

    2013-01-01

    The environmental impact assessment (EIA) has been a tool for decision makers since the enactment of the National Environmental Policy Act (NEPA). Since that time, few analyses have been performed to verify the quality of information and content within EIAs. High quality information within assessments is vital in order for decision makers, stake holders, and the public to understand the potential impact of proposed actions on the ecosystem and wildlife species. Low quality information has been a major cause for litigation and economic loss. Since 1999, wind energy development has seen an exponential growth with unknown levels of impact on wildlife species, in particular bird and bat species. The purpose of this article is to: (1) develop, validate, and apply a quantitative index to review avian/bat assessment quality for wind energy EIAs; and (2) assess the trends and status of avian/bat assessment quality in a sample of wind energy EIAs. This research presents the development and testing of the Avian and Bat Assessment Quality Index (ABAQI), a new approach to quantify information quality of ecological assessments within wind energy development EIAs in relation to avian and bat species based on review areas and factors derived from 23 state wind/wildlife siting guidance documents. The ABAQI was tested through a review of 49 publicly available EIA documents and validated by identifying high variation in avian and bat assessments quality for wind energy developments. Of all the reviewed EIAs, 66% failed to provide high levels of preconstruction avian and bat survey information, compared to recommended factors from state guidelines. This suggests the need for greater consistency from recommended guidelines by state, and mandatory compliance by EIA preparers to avoid possible habitat and species loss, wind energy development shut down, and future lawsuits. - Highlights: ► We developed, validated, and applied a quantitative index to review avian/bat assessment quality

  12. An innovative implementation of LCA within the EIA procedure: Lessons learned from two Wastewater Treatment Plant case studies

    International Nuclear Information System (INIS)

    Larrey-Lassalle, Pyrène; Catel, Laureline; Roux, Philippe; Rosenbaum, Ralph K.; Lopez-Ferber, Miguel; Junqua, Guillaume; Loiseau, Eléonore

    2017-01-01

    Life Cycle Assessment (LCA) has been identified in the literature as a promising tool to increase the performance of environmental assessments at different steps in the Environmental Impact Assessment (EIA) procedure. However, few publications have proposed a methodology for an extensive integration, and none have compared the results with existing EIA conclusions without LCA. This paper proposes a comprehensive operational methodology for implementing an LCA within an EIA. Based on a literature review, we identified four EIA steps that could theoretically benefit from LCA implementation, i.e., (a) the environmental comparison of alternatives, (b) the identification of key impacts, (c) the impact assessment, and (d) the impact of mitigation measures. For each of these steps, an LCA was implemented with specific goal and scope definitions that resulted in a specific set of indicators. This approach has been implemented in two contrasting Wastewater Treatment Plant (WWTP) projects and compared to existing EIA studies. The results showed that the two procedures, i.e., EIAs with or without inputs from LCA, led to different conclusions. The environmental assessments of alternatives and mitigation measures were not carried out in the original studies and showed that other less polluting technologies could have been chosen. Regarding the scoping step, the selected environmental concerns were essentially different. Global impacts such as climate change or natural resource depletion were not taken into account in the original EIA studies. Impacts other than those occurring on the project site (off-site impacts) were not assessed, either. All these impacts can be significant compared to those initially considered. On the other hand, unlike current LCA applications, EIAs usually address natural and technological risks and neighbourhood disturbances such as noises or odours, which are very important for the public acceptability of projects. Regarding the impact assessment

  13. Environmental impact assessment (EIA) in development cooperation. A presentation of development challenges and research issues in developing countries and donor agencies

    Energy Technology Data Exchange (ETDEWEB)

    Kristoffersen, Harald

    1997-12-31

    This document discusses some development challenges and research needs related to environmental impact assessment (EIA) in development cooperation. After a general introduction to basic principles of EIA, the document deals with some general conditions for EIA in developing countries and in donor agencies. Through a presentation of experiences with EIA from selected donor agencies (with emphasis on NORAD) the report ends up with focusing on some research issues that may come up with recommendations for improving EIA practices in developing countries and donor agencies. 37 refs., 6 figs., 3 refs.

  14. A generic approach to integrate biodiversity considerations in screening and scoping for EIA

    International Nuclear Information System (INIS)

    Slootweg, Roel; Kolhoff, Arend

    2003-01-01

    The Convention on Biological Diversity (CBD) requires parties to apply environmental impact assessment (EIA) to projects that potentially negatively impact on biodiversity. As members of the International Association of Impact Assessment, the authors have developed a conceptual framework to integrate biodiversity considerations in EIA. By defining biodiversity in terms of composition, structure, and key processes, and by describing the way in which human activities affect these so-called components of biodiversity, it is possible to assess the potential impacts of human activities on biodiversity. Furthermore, the authors have translated this conceptual framework in generic guidelines for screening and scoping in impact assessment. Countries can use these generic guidelines to further operationalise the framework within the existing national procedures for impact assessment. This paper is fully coherent and partly overlapping with the guidelines recently adopted by the CBD, but differs in the sense that it provides more scientific background and is less policy-oriented

  15. Session 3 report stake holder involvement, particularly in the environmental impact assessment (EIA)

    International Nuclear Information System (INIS)

    Sakuma, H.

    2002-01-01

    Owing much to the seating arrangements that allowed a range of Finnish stakeholders and FSC representatives to share eight small tables, discussions were quite active and hence produced the maximum output, given the time limitations. It should, first of all, be acknowledged that the highly interactive format chosen for the Workshop was a success. Session III focused on the Environmental Impact Assessment (EIA) and the involvement of a variety of stakeholders therein. As introduced by a majority of the plenary speakers prior to the discussion, the EIA formed an integral part of the application submitted by Posiva to the Finnish authorities for a decision in principle according to the Nuclear Energy Act. Reflecting the cross-section of stakeholders at each table, a number of different insights were introduced and discussed. At the end of the discussion, a facilitator for each table summarised the discussion and introduced the summaries in a short presentation. (author)

  16. EIA model documentation: Petroleum Market Model of the National Energy Modeling System

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1994-12-30

    The purpose of this report is to define the objectives of the Petroleum Market Model (PMM), describe its basic approach, and provide detail on how it works. This report is intended as a reference document for model analysts, users, and the public. Documentation of the model is in accordance with EIA`s legal obligation to provide adequate documentation in support of its models (Public Law 94-385, section 57.b.2). The PMM models petroleum refining activities, the marketing of products, the production of natural gas liquids and domestic methanol, projects petroleum provides and sources of supplies for meeting demand. In addition, the PMM estimates domestic refinery capacity expansion and fuel consumption.

  17. Simplified Method for Preliminary EIA of WE Installations based on Newtechnology Classification

    DEFF Research Database (Denmark)

    Margheritini, Lucia

    2010-01-01

    The Environmental Impact Assessment (EIA) is an environmental management instrument implemented worldwide. Full scale WECs are expected to be subjects to EIA. The consents application process can be a very demanding for Wave Energy Converters (WECs) developers. The process is possibly aggravated...... by the lack of common frameworks or coordination between authorities and statutory consultees. Improvement of public administration’s efficiency, including licensing procedures is therefore a key of success for the entire sector. It is believed that part of the challenges to an efficient consents application...... process sit in this huge variety of existing technologies as the environmental impacts of WECs are strongly technology dependent, in addition of being location dependent. The present papers aims at introducing a new technology classification based on the expected environmental impact of the devices...

  18. What would a strategic EIA of the Temelin nuclear power plant probably look like

    International Nuclear Information System (INIS)

    Zeman, J.

    2001-01-01

    The Temelin NPP design was not subjected to Environmental Impact Assessment (EIA) in the early 1980s because Czechoslovak legislation did not require this at that time, although a number of various assessments were carried out later. The article is a hypothetical short EIA of the plant, analyzing the environmental, safety, social, and economic risks. It is concluded that as compared to the construction discontinuation option, completion of the plant and its start-up and operation is a variant which is economically feasible. The social and environmental risks of completion of the plant appear acceptable in comparison with the alternatives, which appear to be economically unacceptable and socially and environmentally rather inconvenient. All alternatives are in contrast with the sustainable development concept. (P.A.)

  19. Social Impact Assessment: The lesser sibling in the South African EIA process?

    International Nuclear Information System (INIS)

    Hildebrandt, L.; Sandham, L.A.

    2014-01-01

    Social Impact Assessment has developed as an integral but neglected component of EIA in South Africa since it became mandatory in 1997, and has therefore been referred to as the “orphan” or “lesser sibling” of EIA, as has SIA in the UK and the US. The aim of this paper is to test this claim by reviewing the quality of a sample of SIA reports, and also to establish whether there has been any improvement in quality following the introduction of revised EIA regulations in 2006. The results confirm that SIA can be called “the lesser sibling” due to the weak grades achieved in the quality review, but also reveal that there has been a slight and consistent improvement in quality, most likely driven by best practice considerations in the absence of prescriptive regulations for SIA. Suggestions and recommendations for addressing observed weakness in SIA performance are advanced. - Highlights: • The quality of a sample of SIA reports was evaluated using a review package. • SIA reports received mostly weak grades. • Limited improvement observed from first to second regulatory regime. • Improvements most likely due to best practice considerations

  20. Public hearings for EIAs in post-communist Bulgaria: do they work?

    International Nuclear Information System (INIS)

    Almer, Heather L.; Koontz, Tomas M.

    2004-01-01

    The participatory practices required as part of the established systems of Environmental Impact Assessment (EIA) practiced in the United States, Canada, Western Europe as well as several international agencies are more recent to the countries of Eastern Europe. The most common official forum for citizen participation in EIAs is the public hearing, which can provide important benefits. However, public hearings have been criticized as exhibiting several problems that preclude meaningful citizen input. This research explores three cases of public hearings held for EIA projects in Bulgaria. It argues that the public participation process and the public hearing in particular share the same problems that have been observed in other countries (including the United States and Canada). At the same time, however, Bulgarian public hearings do provide important indirect benefits that can contribute to the capacity for democratic governance and an active civil society. In the face of substantial economic obstacles and dramatic governmental reforms the country has endured since the end of communism, forums such as the public hearing are important means to foster institutional restructuring of a newly democratized country

  1. An Investigation of the Factors Affecting Performance of Environmental Impact Assessment Practices (EIA in Pakistan

    Directory of Open Access Journals (Sweden)

    Shahbaz Hussain

    2015-01-01

    Full Text Available Environmental Impact Assessment is used to detect changes that a proposed project may have on environment. The intent of present study is to investigate impact of institutional capacity and legal framework on performance of Environmental Impact Assessment practices in Pakistan such as screening, scoping and mitigation, environmental management plan and reporting. Sample of 200 EIA professionals have been selected by using random sampling approach from all provinces of Pakistan. Data has been collected through structured questionnaire and analysed by using AMOS 19 (Analysis of Moment Structures software. Results of path analysis indicated that institutional capacity and legal framework have significant direct impact on performance of all EIA practices in Pakistan. Model fit statistics such as GFI, RMR, NFI, IFI, TLI, RFI, CFI indicate the fitness of research model in this context. It has been suggested that there must be relevant and sufficient human resources that can uplift institutional capacity and legal framework must be well implemented because it will lead to enhanced performance of EIA practices in Pakistan.

  2. Pectic enzymes

    NARCIS (Netherlands)

    Benen, J.A.E.; Voragen, A.G.J.; Visser, J.

    2003-01-01

    The pectic enzymes comprise a diverse group of enzymes. They consist of main-chain depolymerases and esterases active on methyl- and acetylesters of galacturonosyl uronic acid residues. The depolymerizing enzymes comprise hydrolases as wel as lyases

  3. Which amphetamine-type stimulants can be detected by oral fluid immunoassays?

    Science.gov (United States)

    Souza, Daniele Z; Boehl, Paula O; Comiran, Eloisa; Prusch, Débora S; Zancanaro, Ivomar; Fuentefria, Alexandre M; Pechansky, Flavio; Duarte, Paulina C A V; De Boni, Raquel B; Fröehlich, Pedro E; Limberger, Renata P

    2012-02-01

    The use of oral fluid for monitoring drug consumption on roads has many advantages over conventional biological fluids; therefore, several immunoassays have been developed for this purpose. In this work, the ability of 3 commercial immunoassays to detect amphetamine-type stimulants (ATSs) in oral fluid was assessed. In addition, it was reviewed the main controlled ATSs available worldwide, as well as the oral fluid immunological screening tests that have been used for identifying ATSs in drivers. The analytical specificity of amphetamine direct enzyme-linked immunosorbent assay (ELISA), methamphetamine direct ELISA (Immunalysis Corporation), and Oral-View saliva multidrug of abuse test (Alfa Scientific Designs) was evaluated using ATS-spiked oral fluid. Legislation and published articles that report the use of immunological screening tests to detect ATS consumption in conductors were reviewed, including the kit's technical information, project reports, police and drug databases. Even at high concentrations, the tested assays were not able to detect methylphenidate, fenproporex, or diethylpropion, controlled ATSs legally marketed in many countries. This evidences the need to develop new kits that enable one to control the misuse of prescription ATSs on roads through oral fluid immunoassays.

  4. Gliadin Detection in Food by Immunoassay

    Science.gov (United States)

    Grant, Gordon; Sporns, Peter; Hsieh, Y.-H. Peggy

    Immunoassays are very sensitive and efficient tests that are commonly used to identify a specific protein. Examples of applications in the food industry include identification of proteins expressed in genetically modified foods, allergens, or proteins associated with a disease, including celiac disease. This genetic disease is associated with Europeans and affects about one in every 200 people in North America. These individuals react immunologically to wheat proteins, and consequently their own immune systems attack and damage their intestines. This disease can be managed if wheat proteins, specifically "gliadins," are avoided in foods.

  5. Statistical approaches to developing a multiplex immunoassay for determining human exposure to environmental pathogens.

    Science.gov (United States)

    Augustine, Swinburne A J; Simmons, Kaneatra J; Eason, Tarsha N; Griffin, Shannon M; Curioso, Clarissa L; Wymer, Larry J; Shay Fout, G; Grimm, Ann C; Oshima, Kevin H; Dufour, Al

    2015-10-01

    There are numerous pathogens that can be transmitted through water. Identifying and understanding the routes and magnitude of exposure or infection to these microbial contaminants are critical to assessing and mitigating risk. Conventional approaches of studying immunological responses to exposure or infection such as Enzyme-Linked Immunosorbent Assays (ELISAs) and other monoplex antibody-based immunoassays can be very costly, laborious, and consume large quantities of patient sample. A major limitation of these approaches is that they can only be used to measure one analyte at a time. Multiplex immunoassays provide the ability to study multiple pathogens simultaneously in microliter volumes of samples. However, there are several challenges that must be addressed when developing these multiplex immunoassays such as selection of specific antigens and antibodies, cross-reactivity, calibration, protein-reagent interferences, and the need for rigorous optimization of protein concentrations. In this study, a Design of Experiments (DOE) approach was used to optimize reagent concentrations for coupling selected antigens to Luminex™ xMAP microspheres for use in an indirect capture, multiplex immunoassay to detect human exposure or infection from pathogens that are potentially transmitted through water. Results from Helicobacter pylori, Campylobacter jejuni, Escherichia coli O157:H7, and Salmonella typhimurium singleplexes were used to determine the mean concentrations that would be applied to the multiplex assay. Cut-offs to differentiate between exposed and non-exposed individuals were determined using finite mixed modeling (FMM). The statistical approaches developed facilitated the detection of Immunoglobulin G (IgG) antibodies to H. pylori, C. jejuni, Toxoplasma gondii, hepatitis A virus, rotavirus and noroviruses (VA387 and Norwalk strains) in fifty-four diagnostically characterized plasma samples. Of the characterized samples, the detection rate was 87.5% for H

  6. Magnetic Electrochemical Immunoassays with Quantum Dot Labels for Detection of Phosphorylated Acetylcholinesterase in Plasma

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Hua; Wang, Jun; Timchalk, Charles; Lin, Yuehe

    2008-11-01

    A new magnetic electrochemical immunoassay has been developed as a tool for biomonitoring exposures to organophosphate (OP) compounds, e.g., insecticides and chemical nerve agents, by directly detecting organophosphorylated acetylcholinesterase (OP-AChE). This immunoassay uniquely incorporates highly efficient magnetic separation with ultrasensitive square wave voltammetry (SWV) analysis with quantum dots (QDs) as labels. A pair of antibodies was used to achieve the specific recognition of OP-AChE that was prepared with paraoxon as an OP model agent. Antiphosphoserine polyclonal antibodies were anchored on amorphous magnetic particles preferably chosen to capture OP-AChE from the sample matrixes by binding their phosphoserine moieties that were exposed through unfolding the protein adducts. This was validated by electrochemical examinations and enzyme-linked immunosorbent assays. Furthermore, antihuman AChE monoclonal antibodies were labeled with cadmium-source QDs to selectively recognize the captured OP-AChE, as characterized by transmission electron microscopy. The subsequent electrochemical SWV analysis of the cadmium component released by acid from the coupled QDs was conducted on disposable screen-printed electrodes. Experimental results indicated that the SWV-based immunoassays could yield a linear response over a broad concentration range of 0.3-300 ng/mL OP-AChE in human plasma with a detection limit of 0.15 ng/mL. Such a novel electrochemical immunoassay holds great promise as a simple, selective, sensitive, and field-deployable tool for the effective biomonitoring and diagnosis of potential exposures to nerve agents and pesticides.

  7. Multiplexed electrochemical immunoassay of biomarkers using metal sulfide quantum dot nanolabels and trifunctionalized magnetic beads.

    Science.gov (United States)

    Tang, Dianping; Hou, Li; Niessner, Reinhard; Xu, Mingdi; Gao, Zhuangqiang; Knopp, Dietmar

    2013-08-15

    A novel multiplexed stripping voltammetric immunoassay protocol was designed for the simultaneous detection of multiple biomarkers (CA 125, CA 15-3, and CA 19-9 used as models) using PAMAM dendrimer-metal sulfide quantum dot (QD) nanolabels as distinguishable signal tags and trifunctionalized magnetic beads as an immunosensing probe. The probe was prepared by means of co-immobilization of primary monoclonal anti-CA 125, anti-CA 15-3 and anti-CA 19-9 antibodies on a single magnetic bead. The PAMAM dendrimer-metal sulfide QD nanolabels containing CdS, ZnS and PbS were synthesized by using in situ synthesis method, which were utilized for the labeling of polyclonal rabbit anti-CA 125, anti-CA 15-3 and anti-CA 19-9 detection antibodies, respectively. A sandwich-type immunoassay format was adopted for the simultaneous determination of target biomarkers in a low-binding microtiter plate. The subsequent anodic stripping voltammetric analysis of cadmium, zinc, and lead components released by acid from the corresponding QD nanolabels was conducted at an in situ prepared mercury film electrode based on the difference of peak potentials. Experimental results indicated that the multiplexed immunoassay enabled the simultaneous detection of three cancer biomarkers in a single run with wide dynamic ranges of 0.01-50 U mL(-1) and detection limits (LODs) of 0.005 U mL(-1). Intra-assay and inter-assay coefficients of variation (CVs) were less than 7.2% and 10.4%, respectively. No significant differences at the 0.05 significance level were encountered in the analysis of 10 clinical serum specimens between the multiplexed immunoassay and a commercially available enzyme-linked immunosorbent assay (ELISA). Copyright © 2013 Elsevier B.V. All rights reserved.

  8. Human CCDC47 sandwich immunoassay development with electrochemiluminescence technology.

    Science.gov (United States)

    Wu, Wenfang S; Zhu, Liang; Patil, Saurabh; Gokul, Karthiga; Reilly, Sean; Chan, Joyce; Tekumalla, Poornima; Vishnudas, Vivek; Kiebish, Michael A; Sarangarajan, Rangaprasad; Akmaev, Viatcheslav R; Kellogg, Mark D; Narain, Niven R

    2018-01-01

    Coiled-Coil Domain Containing 47 (CCDC47) is an endoplasmic reticulum (ER) transmembrane protein involved in calcium signaling through utilization of its calcium binding-acidic luminal domain. CCDC47 also interacts with ERAD (endoplasmic reticulum-associated degradation) complex and is involved in ER stress relief. In this report, we developed human CCDC47 monoclonal antibodies and a sandwich immunoassay for CCDC47 measurement in biological matrices. Specificity of developed antibodies were confirmed by immunoblot and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis of immunoprecipitated cell lysates. To achieve high analytical sensitivity, traditional colorimetric enzyme-linked immunosorbent assay (ELISA) and electrochemiluminescence (ECL) technology were compared, and 3 logs of increased sensitivity was observed with the use of ECL. A CCDC47 sandwich ECL assay was subsequently developed and performances evaluated for calibration curves, precision and accuracy, as well as selectivity and interferences for sample measurement. Sample stability was also characterized for freeze/thaw cycles and short/long term storage conditions. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Nanoparticle-based immunosensors and immunoassays for aflatoxins

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Xu; Niessner, Reinhard [Institute of Hydrochemistry and Chair of Analytical Chemistry, Technische Universität München, Marchioninistrasse 17, D-81377 München (Germany); Tang, Dianping [Key Laboratory of Analysis and Detection for Food Safety, MOE & Fujian Province, Department of Chemistry, Fuzhou University, Fuzhou 350108 (China); Knopp, Dietmar, E-mail: dietmar.knopp@ch.tum.de [Institute of Hydrochemistry and Chair of Analytical Chemistry, Technische Universität München, Marchioninistrasse 17, D-81377 München (Germany)

    2016-03-17

    Aflatoxins are naturally existing mycotoxins produced mainly by Aspergillus flavus and Aspergillus parasiticus, present in a wide range of food and feed products. Because of their extremely high toxicity and carcinogenicity, strict control of maximum residue levels of aflatoxins in foodstuff is set by many countries. In daily routine, different chromatographic methods are used almost exclusively. As supplement, in several companies enzyme immunoassay-based sample testing as primary screening is performed. Recently, nanomaterials such as noble metal nanoparticles, magnetic particles, carbon nanomaterials, quantum dots, and silica nanomaterials are increasingly utilized for aflatoxin determination to improve the sensitivity and simplify the detection. They are employed either as supports for the immobilization of biomolecules or as electroactive or optical labels for signal transduction and amplification. Several nanoparticle-based electrochemical, piezoelectric, optical, and immunodipstick assays for aflatoxins have been developed. In this review, we summarize these recent advances and illustrate novel concepts and promising applications in the field of food safety. - Highlights: • Novel concepts and promising applications of nanoparticle-based immunological methods for the determination of aflatoxins. • Inclusion of most important nanomaterials and hybrid nanostructures. • Inclusion of electrochemical, optical and mass-sensitive biosensors as well as optical and immunochromatographic assays.

  10. Nanoparticle-based immunosensors and immunoassays for aflatoxins

    International Nuclear Information System (INIS)

    Wang, Xu; Niessner, Reinhard; Tang, Dianping; Knopp, Dietmar

    2016-01-01

    Aflatoxins are naturally existing mycotoxins produced mainly by Aspergillus flavus and Aspergillus parasiticus, present in a wide range of food and feed products. Because of their extremely high toxicity and carcinogenicity, strict control of maximum residue levels of aflatoxins in foodstuff is set by many countries. In daily routine, different chromatographic methods are used almost exclusively. As supplement, in several companies enzyme immunoassay-based sample testing as primary screening is performed. Recently, nanomaterials such as noble metal nanoparticles, magnetic particles, carbon nanomaterials, quantum dots, and silica nanomaterials are increasingly utilized for aflatoxin determination to improve the sensitivity and simplify the detection. They are employed either as supports for the immobilization of biomolecules or as electroactive or optical labels for signal transduction and amplification. Several nanoparticle-based electrochemical, piezoelectric, optical, and immunodipstick assays for aflatoxins have been developed. In this review, we summarize these recent advances and illustrate novel concepts and promising applications in the field of food safety. - Highlights: • Novel concepts and promising applications of nanoparticle-based immunological methods for the determination of aflatoxins. • Inclusion of most important nanomaterials and hybrid nanostructures. • Inclusion of electrochemical, optical and mass-sensitive biosensors as well as optical and immunochromatographic assays.

  11. Comparison of radioimmunological and enzyme-immunological methods of determination in thyroid function studies

    International Nuclear Information System (INIS)

    Arnold, L.P.

    1984-01-01

    During the study described here parallel investigations were carried out using radio-immuno-assays (RIA) and enzyme-immuno-assays (EIA) in order to assess the quality and diagnostic value of both these techniques as well as to weigh up their relative advantages and disadvantages in connection with thyroid function studies. The following comparisons were made: T4 RIA versus T4 EIA, T3 RIA versus T3 EIA, T3 uptake versus thyroxine binding capacity, total balance of free hormone bonds. Correlation coefficients of 0.982 for T4 and 0.989 for T3 proved that the test results obtained using either RIA or EIA were in fairly good agreement. Less satisfactory correlation was suggested by a coefficient of 0.807 between the tests for thyroxine binding capacity and T3 uptake, which was even seen to decrease with increasing binding capacity so that the greatest deviations were observed in hyperthyroidism. The same holds true for the free T4 index and total balance, where the correlation coefficient is altered in a similar way by the inclusion of a thyroxine binding capacity EIA. Better agreement was achieved by optimal adjustment of the defined diagnostic ranges for hypothyroidism, euthyroidism and hyperthyroidism. The EIA results were biassed in the presence of hyperbilirubinemia. Intra-assay abd inter-assay veriations were analysed and found to range from 8.1 to 16.9% and 13.6 to 28.1%, respectively. EIA offers advantages over RIA inasmuch as it is free from radioactivity and does not require any special safety measures, although it was judged considerably less favourable than RIA in terms of sensitivity. (TRV) [de

  12. Serotype sensitivity of a lateral flow immunoassay for cryptococcal antigen.

    Science.gov (United States)

    Gates-Hollingsworth, Marcellene A; Kozel, Thomas R

    2013-04-01

    To meet the needs of a global community, an immunoassay for cryptococcal antigen (CrAg) must have high sensitivity for CrAg of all major serotypes. A new immunoassay for CrAg in lateral flow format was evaluated and found to have a high sensitivity for detection of serotypes A, B, C, and D.

  13. Microarray-integrated optoelectrofluidic immunoassay system.

    Science.gov (United States)

    Han, Dongsik; Park, Je-Kyun

    2016-05-01

    A microarray-based analytical platform has been utilized as a powerful tool in biological assay fields. However, an analyte depletion problem due to the slow mass transport based on molecular diffusion causes low reaction efficiency, resulting in a limitation for practical applications. This paper presents a novel method to improve the efficiency of microarray-based immunoassay via an optically induced electrokinetic phenomenon by integrating an optoelectrofluidic device with a conventional glass slide-based microarray format. A sample droplet was loaded between the microarray slide and the optoelectrofluidic device on which a photoconductive layer was deposited. Under the application of an AC voltage, optically induced AC electroosmotic flows caused by a microarray-patterned light actively enhanced the mass transport of target molecules at the multiple assay spots of the microarray simultaneously, which reduced tedious reaction time from more than 30 min to 10 min. Based on this enhancing effect, a heterogeneous immunoassay with a tiny volume of sample (5 μl) was successfully performed in the microarray-integrated optoelectrofluidic system using immunoglobulin G (IgG) and anti-IgG, resulting in improved efficiency compared to the static environment. Furthermore, the application of multiplex assays was also demonstrated by multiple protein detection.

  14. The development and validation of an immunoassay for the measurement of anti-thymidine phosphorylase antibodies in mouse and dog sera

    OpenAIRE

    Gasson, C; Levene, M; Bax, BE

    2013-01-01

    Erythrocyte encapsulated thymidine phosphorylase (EE-TP) is under development as an enzyme replacement therapy for mitochondrial neurogastrointestinal encephalomyopathy (MNGIE), a fatal metabolic disorder resulting from an inherited deficiency of the enzyme thymidine phosphorylase. We report here the development and validation of a sensitive electrochemiluminescent (ECL) bridging immunoassay to support Good Laboratory Practice (GLP)-compliant preclinical safety studies of EE-TP in the mouse a...

  15. A study on multifrequency scintillations near the EIA crest of the Indian zone

    Science.gov (United States)

    Chakraborty, S. K.; Chatterjee, S.; Jana, Debasis

    2017-10-01

    Occurrence features of ionospheric scintillations at S band (2492.028 MHz) are reported for the first time. The same have been explored in the context of scintillations at VHF (250.650 MHz) and L5 (1176.45 MHz) bands. Observations were carried out during the period April-December, 2015 at Raja Peary Mohan College Centre (RPMC: 22.66° N, 88.4° E), located near the equatorial ionization anomaly (EIA) crest of the Indian longitude zone. Mostly weak (theory of evolution, structure and dynamics of electron density irregularities in the low latitude region.

  16. Irregular-shaped platinum nanoparticles as peroxidase mimics for highly efficient colorimetric immunoassay.

    Science.gov (United States)

    Gao, Zhuangqiang; Xu, Mingdi; Hou, Li; Chen, Guonan; Tang, Dianping

    2013-05-07

    Enzyme-linked immunosorbent assay (ELISA) methods based on natural enzyme-labeled probes have been applied in the immunoassays, but most have some inevitable limitations (e.g. harsh preparation, purification and storage) and are unsuitable for routine use. Herein we synthesized a new class of irregular-shaped platinum nanoparticles (ISPtNP) with a mean length of 7.0 nm and a narrowing width from 2.0 to 5.0 nm along the longitudinal axes, which were utilized as peroxidase-like mimics for the development of colorimetric immunoassays. Compared with bioactive horseradish peroxidase (HRP), the synthesized ISPtNP exhibited a low Km value (~0.12 mM) and a high Kcat value (~2.27×10(4)s(-1)) for 3,3',5,5'-tetramethylbenzidine (TMB) with strong thermal stability and pH tolerance. The catalytic mechanism of the ISPtNP toward TMB/H2O2 was for the first time discussed and deliberated in this work. Based on a sandwich-type assay format, two types of colorimetric immunoassay protocols were designed and developed for the detection of rabbit IgG (RIgG, as a model) by using the synthesized ISPtNP and conventional HRP as the labeling of detection antibodies, respectively. Similar detection limits (LODs) of 2.5 ng mL(-1) vs. 1.0 ng mL(-1) were obtained toward RIgG with the ISPtNP labeling compared to HRP format. Intra- and inter-assay coefficients of variation were less than 13%. Importantly, the ISPtNP-based assay system could be suitable for use in a mass production of miniaturized lab-on-a-chip devices and open new opportunities for protein diagnostics and biosecurity. Copyright © 2013 Elsevier B.V. All rights reserved.

  17. UK and Italian EIA systems: A comparative study on management practice and performance in the construction industry

    Energy Technology Data Exchange (ETDEWEB)

    Bassi, Andrea, E-mail: ab395@bath.co.uk [University of Bath, Faculty of Engineering and Design, Claverton Down, Bath BA2 7AY (United Kingdom); Howard, Robert, E-mail: robhoward@constcom.demon.co.uk [Construction Communications, 8 Cotton& #x27; s Field, Dry Drayton, Cambridge CB23 8DG (United Kingdom); Geneletti, Davide, E-mail: davide.geneletti@ing.unitn.it [Sustainability Science Program, Harvard University, 79 JFK Street, Cambridge, MA 02138 (United States); Dept. of Civil and Environmental Engineering, University of Trento, Via Mesiano, 77 38123 Trento (Italy); Ferrari, Simone, E-mail: simone.ferrari@polimi.it [Dept. BEST, Building Environment Science and Technology, Politecnico di Milano, Via Bonardi, 3 20133 Milano (Italy)

    2012-04-15

    This study evaluates and contrasts the management practice and the performance that characterise Environmental Impact Assessments (EIA) in Italy and in the UK. The methodology relies on the investigation of six carefully selected case studies, critically reviewed by referring to EIA and project design information, as well as collecting the opinion of key project participants. The study focuses on the construction industry and on specific key sectors like infrastructure for transport and renewable energy and commercial and tourism development. A main term of reference for the analyses has been established by critically reviewing international literature so as to outline common good practice, requirements for the enhancement of sustainability principles and typically incurred drawbacks. The proposed approach enhances transfer of knowledge and of experiences between the analyzed contexts and allows the provision of guidelines for practitioners. Distinctive differences between the UK and the Italian EIA systems have been detected for pivotal phases and elements of EIA, like screening, scoping, analysis of alternatives and of potential impacts, definition of mitigation strategies, review, decision making, public participation and follow up. - Highlights: Black-Right-Pointing-Pointer The Italian and the UK Environmental Impact Assessment systems are compared. Black-Right-Pointing-Pointer The research is centred on the construction industry. Black-Right-Pointing-Pointer Issues and shortcomings are analysed by investigating six case studies. Black-Right-Pointing-Pointer Integration of EIA with sustainability principles is appraised. Black-Right-Pointing-Pointer General guidelines are provided to assist practitioners in the two national contexts.

  18. UK and Italian EIA systems: A comparative study on management practice and performance in the construction industry

    International Nuclear Information System (INIS)

    Bassi, Andrea; Howard, Robert; Geneletti, Davide; Ferrari, Simone

    2012-01-01

    This study evaluates and contrasts the management practice and the performance that characterise Environmental Impact Assessments (EIA) in Italy and in the UK. The methodology relies on the investigation of six carefully selected case studies, critically reviewed by referring to EIA and project design information, as well as collecting the opinion of key project participants. The study focuses on the construction industry and on specific key sectors like infrastructure for transport and renewable energy and commercial and tourism development. A main term of reference for the analyses has been established by critically reviewing international literature so as to outline common good practice, requirements for the enhancement of sustainability principles and typically incurred drawbacks. The proposed approach enhances transfer of knowledge and of experiences between the analyzed contexts and allows the provision of guidelines for practitioners. Distinctive differences between the UK and the Italian EIA systems have been detected for pivotal phases and elements of EIA, like screening, scoping, analysis of alternatives and of potential impacts, definition of mitigation strategies, review, decision making, public participation and follow up. - Highlights: ► The Italian and the UK Environmental Impact Assessment systems are compared. ► The research is centred on the construction industry. ► Issues and shortcomings are analysed by investigating six case studies. ► Integration of EIA with sustainability principles is appraised. ► General guidelines are provided to assist practitioners in the two national contexts.

  19. The influence of actor capacities on EIA system performance in low and middle income countries —Cases from Georgia and Ghana

    Energy Technology Data Exchange (ETDEWEB)

    Kolhoff, Arend J., E-mail: akolhoff@eia.nl [Netherlands Commission for Environmental Assessment, P.O. Box 2345, 3500 GH Utrecht (Netherlands); Runhaar, Hens A.C., E-mail: H.A.C.Runhaar@uu.nl [Environmental Studies and Policy, Copernicus Institute of Sustainable Development, Utrecht University, Utrecht (Netherlands); Forest and Nature Conservation Policy Group, Wageningen University and Research Centre, Wageningen (Netherlands); Gugushvili, Tamar, E-mail: gugushvili.tamuna@gmail.com [Netherlands Commission for Environmental Assessment, P.O. Box 2345, 3500 GH Utrecht (Netherlands); Sonderegger, Gabi, E-mail: gabi.sonderegger@gmail.com [Netherlands Commission for Environmental Assessment, P.O. Box 2345, 3500 GH Utrecht (Netherlands); Van der Leest, Bart, E-mail: bartvanderleest@hotmail.com [Netherlands Commission for Environmental Assessment, P.O. Box 2345, 3500 GH Utrecht (Netherlands); Driessen, Peter P.J., E-mail: p.driessen@uu.nl [Innovation and Environmental Sciences, Copernicus Institute of Sustainable Development, Utrecht University, Utrecht (Netherlands)

    2016-02-15

    In this paper, we aim to better understand the factors that contribute to the substantive performance of EIA systems in low and middle income countries. Substantive performance is defined as the extent to which the EIA process contributes to the EIA objectives for the long term, namely environmental protection or, even more ambitious, sustainable development. We have therefore developed a conceptual model in which we focus on the key actors in the EIA system, the proponent and the EIA authority and their level of ownership as a key capacity to measure their performance, and we distinguish procedural performance and some contextual factors. This conceptual model is then verified and refined for the EIA phase and the EIA follow-up phase (permitting, monitoring and enforcement) by means of 12 case studies from Ghana (four cases) and Georgia (eight cases), both lower–middle income countries. We observe that in most cases the level of substantive performance increases during the EIA phase but drops during the EIA follow-up phase, and as a result only five out of 12 operational cases are in compliance with permit conditions or national environmental standards. We conclude, firstly that ownership of the proponent is the most important factor explaining the level of substantive performance; the higher the proponent's level of ownership the higher the level of substantive performance. The influence of the EIA authority on substantive performance is limited. Secondly, the influence of procedural performance on substantive performance seems less important than expected in the EIA phase but more important during the EIA follow-up phase. In order to improve substantive performance we learned two lessons. Firstly, increasing the proponent's level of ownership seems obvious, but direct change is probably difficult. However, where international finance institutes are involved they can increase ownership. Despite the limited influence of the EIA authority, a proactive

  20. Microretroreflector-sedimentation immunoassays for pathogen detection.

    Science.gov (United States)

    Garvey, Gavin; Shakarisaz, David; Ruiz-Ruiz, Federico; Hagström, Anna E V; Raja, Balakrishnan; Pascente, Carmen; Kar, Archana; Kourentzi, Katerina; Rito-Palomares, Marco; Ruchhoeft, Paul; Willson, Richard C

    2014-09-16

    Point-of-care detection of pathogens is medically valuable but poses challenging trade-offs between instrument complexity and clinical and analytical sensitivity. Here we introduce a diagnostic platform utilizing lithographically fabricated micron-scale forms of cubic retroreflectors, arguably one of the most optically detectable human artifacts, as reporter labels for use in sensitive immunoassays. We demonstrate the applicability of this novel optical label in a simple assay format in which retroreflector cubes are first mixed with the sample. The cubes are then allowed to settle onto an immuno-capture surface, followed by inversion for gravity-driven removal of nonspecifically bound cubes. Cubes bridged to the capture surface by the analyte are detected using inexpensive, low-numerical aperture optics. For model bacterial and viral pathogens, sensitivity in 10% human serum was found to be 10(4) bacterial cells/mL and 10(4) virus particles/mL, consistent with clinical utility.

  1. Diverse patterns of recognition of hepatitis C virus core and nonstructural antigens by antibodies present in Egyptian cancer patients and blood donors

    NARCIS (Netherlands)

    Attia, M. A.; Zekri, A. R.; Goudsmit, J.; Boom, R.; Khaled, H. M.; Mansour, M. T.; de Wolf, F.; el-Din, H. M.; Sol, C. J.

    1996-01-01

    Serum samples from 429 cancer patients, 82 unpaid blood donors, and 74 paid blood donors were tested for hepatitis C virus (HCV) markers in two commercially available enzyme immunoassays (EIAs). A total of 229 of 429 (53.4%) cancer patients were positive by the two EIAs. A total of 34 of 156 (21.8%)

  2. From consultation to deliberation? Tracing deliberative norms in EIA frameworks in Swedish roads planning

    International Nuclear Information System (INIS)

    Isaksson, Karolina; Richardson, Tim; Olsson, Krister

    2009-01-01

    This paper presents the results of an analysis of deliberative norms in the framework for Environmental Impact Assessment (EIA) in roads planning in Sweden. The more specific question is how this framework has responded to the shift towards more deliberative approaches to planning and decision making, advocated in planning theory and policy literature over the last decade. The analysis, which compares the current framework and guidance with an earlier iteration, identifies a shift towards deliberation; deliberative norms are present, and even dominate recent guidance. However, an instrumental norm permeates both the former and the current guidance, suggesting that even as a language of consultation is replaced by one of deliberation, the intention remains to secure and legitimise a smooth development pathway. Evidence from interviews with professionals working in the Swedish EIA system highlights the difficulties of navigating these uncertainties in practice. By opening up critical analysis of deliberative norms as they shape the conditions for practice, this study contributes to the continuous development of planning practice, by supporting a more normatively reflexive approach to framework-design.

  3. EIA model documentation: Petroleum market model of the national energy modeling system

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1995-12-28

    The purpose of this report is to define the objectives of the Petroleum Market Model (PMM), describe its basic approach, and provide detail on how it works. This report is intended as a reference document for model analysts, users, and the public. Documentation of the model is in accordance with EIA`s legal obligation to provide adequate documentation in support of its models. The PMM models petroleum refining activities, the marketing of petroleum products to consumption regions, the production of natural gas liquids in gas processing plants, and domestic methanol production. The PMM projects petroleum product prices and sources of supply for meeting petroleum product demand. The sources of supply include crude oil, both domestic and imported; other inputs including alcohols and ethers; natural gas plant liquids production; petroleum product imports; and refinery processing gain. In addition, the PMM estimates domestic refinery capacity expansion and fuel consumption. Product prices are estimated at the Census division level and much of the refining activity information is at the Petroleum Administration for Defense (PAD) District level.

  4. An INMARSAT-C goes, and EIA485 hybrid communication system for global experiment control

    Energy Technology Data Exchange (ETDEWEB)

    Reynolds, R.B.; Behrens, W.

    1995-08-01

    The Atmosphere Radiation Measurement (ARM) program is constructing and radiation monitoring instrument that will be deployed at several island sites in the Tropical Western Pacific Ocean. The atmospheric radiation and cloud systems (ARCS) must operate the minimal maintenance for decades, producing and storing many megabytes of data per day. A reliable global communication system is an essential element of the ARCS design. It must provide immediate direct access to the equipment. High availability and reasonably low cost are essential for this very long-term deployment. Providing executive management and control, the communication system incorporates several special technologies to meet its requirements: (a) INMARSAT-C provides a simple two-way messaging capability. (b) Built-in GPS in the INMARSAT-C radio provides an accurate time standard. (c) The NOAA GOES satellite provides a one-way link for hourly health and basic measurement data. (d) An EIA485 local area network provides a digital link throughout the installation. (e) Node units located conveniently throughout the ARCS and linked by the EIA485 network allow an almost unlimited flexibility and expandability. (f) A robust packet protocol ensures message security and accuracy. The complete communication, control, and data acquisition system is described in detail. The ability of the system to adapt to other physical configurations (telephone, UHF, telnet) is discussed.

  5. Greenpeace and the EIA process with special reference to the siting of a repository for spent nuclear fuel

    International Nuclear Information System (INIS)

    Fredriksson, A.

    1995-01-01

    The author argues as follows: The legislation which currently regulates EIS, does not give any party other than the proponent the possibility of exercising influence over the EIA. Therefore, an extensive review and amendment of the existing legislation is necessary if the general public is to participate in the process. This should be regarded as necessary, if SKB is to gain general acceptance of its work. Work on developing a suitable method and identifying a suitable site is going on, but is not conducted in such a manner that it gives the general public an adequate possibility of participating. Furthermore, decision-makers in the candidate municipalities are not being given insight into the work. A specially established EIA authority, charged with the task of supervising the EIA, may lead to that the problems will be investigated in a better way, that can lead to the selection of the best method as well as the best site

  6. Mass Spectrometric Immunoassays in Characterization of Clinically Significant Proteoforms

    Directory of Open Access Journals (Sweden)

    Olgica Trenchevska

    2016-03-01

    Full Text Available Proteins can exist as multiple proteoforms in vivo, as a result of alternative splicing and single-nucleotide polymorphisms (SNPs, as well as posttranslational processing. To address their clinical significance in a context of diagnostic information, proteoforms require a more in-depth analysis. Mass spectrometric immunoassays (MSIA have been devised for studying structural diversity in human proteins. MSIA enables protein profiling in a simple and high-throughput manner, by combining the selectivity of targeted immunoassays, with the specificity of mass spectrometric detection. MSIA has been used for qualitative and quantitative analysis of single and multiple proteoforms, distinguishing between normal fluctuations and changes related to clinical conditions. This mini review offers an overview of the development and application of mass spectrometric immunoassays for clinical and population proteomics studies. Provided are examples of some recent developments, and also discussed are the trends and challenges in mass spectrometry-based immunoassays for the next-phase of clinical applications.

  7. Enzyme Informatics

    Science.gov (United States)

    Alderson, Rosanna G.; Ferrari, Luna De; Mavridis, Lazaros; McDonagh, James L.; Mitchell, John B. O.; Nath, Neetika

    2012-01-01

    Over the last 50 years, sequencing, structural biology and bioinformatics have completely revolutionised biomolecular science, with millions of sequences and tens of thousands of three dimensional structures becoming available. The bioinformatics of enzymes is well served by, mostly free, online databases. BRENDA describes the chemistry, substrate specificity, kinetics, preparation and biological sources of enzymes, while KEGG is valuable for understanding enzymes and metabolic pathways. EzCatDB, SFLD and MACiE are key repositories for data on the chemical mechanisms by which enzymes operate. At the current rate of genome sequencing and manual annotation, human curation will never finish the functional annotation of the ever-expanding list of known enzymes. Hence there is an increasing need for automated annotation, though it is not yet widespread for enzyme data. In contrast, functional ontologies such as the Gene Ontology already profit from automation. Despite our growing understanding of enzyme structure and dynamics, we are only beginning to be able to design novel enzymes. One can now begin to trace the functional evolution of enzymes using phylogenetics. The ability of enzymes to perform secondary functions, albeit relatively inefficiently, gives clues as to how enzyme function evolves. Substrate promiscuity in enzymes is one example of imperfect specificity in protein-ligand interactions. Similarly, most drugs bind to more than one protein target. This may sometimes result in helpful polypharmacology as a drug modulates plural targets, but also often leads to adverse side-effects. Many cheminformatics approaches can be used to model the interactions between druglike molecules and proteins in silico. We can even use quantum chemical techniques like DFT and QM/MM to compute the structural and energetic course of enzyme catalysed chemical reaction mechanisms, including a full description of bond making and breaking. PMID:23116471

  8. The fabrication of magnetic particle-based chemiluminescence immunoassay for human epididymis protein-4 detection in ovarian cancer.

    Science.gov (United States)

    Fu, Xiaoling; Liu, Yangyang; Qiu, Ruiyun; Foda, Mohamed F; Zhang, Yong; Wang, Tao; Li, Jinshan

    2018-03-01

    The magnetic particles have a significant influence on the immunoassay detection and cancer therapy. Herein, the chemiluminescence immunoassay combined with the magnetic particles (MPCLIA) was presented for the clinical determination and analysis of human epididymis protein 4 (HE4) in the human serum. Under the optimized experiment conditions, the secure MPCLIA method can detect HE4 in the broader range of 0-1000 pmol/L, with a lower detection limit of 1.35 pmol/L. The satisfactory recovery rate of the method in the serum ranged from 83.62% to 105.10%, which was well within the requirement of clinical analysis. Moreover, the results showed the good correlation with enzyme-linked immunosorbent assay (ELISA), with the correlation coefficient of 0.9589. This proposed method has been successfully applied to the clinical determination of HE4 in the human serum.

  9. An analysis framework for characterizing and explaining development of EIA legislation in developing countries-Illustrated for Georgia, Ghana and Yemen

    Energy Technology Data Exchange (ETDEWEB)

    Kolhoff, Arend J., E-mail: akolhoff@eia.nl [Netherlands Commission for Environmental Assessment, P.O. Box 2345, 3500 GH Utrecht (Netherlands); Driessen, Peter P.J., E-mail: p.driessen@uu.nl [Copernicus Institute of Sustainable Development, Utrecht University, P.O. Box 80115, 3508 TC (Netherlands); Runhaar, Hens A.C., E-mail: h.a.c.runhaar@uu.nl [Copernicus Institute of Sustainable Development, Utrecht University, P.O. Box 80115, 3508 TC (Netherlands)

    2013-01-15

    Actors in the field of international development co-operation supporting the development of EIA legislation in developing countries often do not achieve the results envisaged. The performance of EIA in these countries often remains weak. One reason, we assume, is that often those actors support the establishment of overly ambitious EIA legislation that cannot achieve its objectives in the light of constraining contexts. To provide more effective support we need to better understand the enabling and constraining contextual factors that influence the development of EIA legislation and to which support actors should align itself. In this article a new analysis framework for classifying, characterizing and explaining the development of EIA legislation is described, measured in terms of ambition levels. Ambitions are defined as intentions the EIA authorities aim to fulfill, expressed in formal EIA legislation. Three country cases, Yemen, Georgia and Ghana are used to illustrate the usefulness of our framework and as a first test to refine the framework. We have formulated the following five hypotheses that complement and refine our analysis framework. One, EIA legislation may develop multilinearly in terms of ambition levels. Two, ambitions in EIA legislation seem to be influenced to a great extent by the power and capacity of, on the one hand, the environmental authorities supporting EIA and, on the other hand, the sector authorities hindering the development of EIA. Three, the political system is the most important context factor influencing the rules of policy-making and the power of the different actors involved. Four, the importance of context factors on the development of ambitions is dependent on the phase of EIA system development. Five, some ambitions seem to be influenced by particular factors; for instance the ambitions for the object of study seem to be influenced by the level of environmental awareness of the sector ministries and parliament. The analysis

  10. Detection of narcotics with an immunoassay film badge

    International Nuclear Information System (INIS)

    Lukens, H.R.

    1993-01-01

    Efficient personnel performance, a major requirement for a safe nuclear industry, is jeopardized where personnel use narcotics. However, detection of narcotics at nuclear plants is a challenge. The unique specificity and sensitivity of an immunoassay has been implemented in the form of a small, dry immunoassay film badge (IFB) for the detection of vapors emitted by narcotics. The device is suitable as an area monitor, and its characteristics are suitable for use as a breath monitor for the detection of drug use

  11. Critical appraisal of four IL-6 immunoassays.

    Directory of Open Access Journals (Sweden)

    Dana K Thompson

    Full Text Available Interleukin-6 (IL-6 contributes to numerous inflammatory, metabolic, and physiologic pathways of disease. We evaluated four IL-6 immunoassays in order to identify a reliable assay for studies of metabolic and physical function. Serial plasma samples from intravenous glucose tolerance tests (IVGTTs, with expected rises in IL-6 concentrations, were used to test the face validity of the various assays.IVGTTs, administered to 14 subjects, were performed with a single infusion of glucose (0.3 g/kg body mass at time zero, a single infusion of insulin (0.025 U/kg body mass at 20 minutes, and frequent blood collection from time zero to 180 minutes for subsequent Il-6 measurement. The performance metrics of four IL-6 detection methods were compared: Meso Scale Discovery immunoassay (MSD, an Invitrogen Luminex bead-based multiplex panel (LX, an Invitrogen Ultrasensitive Luminex bead-based singleplex assay (ULX, and R&D High Sensitivity ELISA (R&D. IL-6 concentrations measured with MSD, R&D and ULX correlated with each other (Pearson Correlation Coefficients r = 0.47-0.94, p<0.0001 but only ULX correlated (r = 0.31, p = 0.0027 with Invitrogen Luminex. MSD, R&D, and ULX, but not LX, detected increases in IL-6 in response to glucose. All plasma samples were measurable by MSD, while 35%, 1%, and 4.3% of samples were out of range when measured by LX, ULX, and R&D, respectively. Based on representative data from the MSD assay, baseline plasma IL-6 (0.90 ± 0.48 pg/mL increased significantly as expected by 90 minutes (1.29 ± 0.59 pg/mL, p = 0.049, and continued rising through 3 hours (4.25 ± 3.67 pg/mL, p = 0.0048.This study established the face validity of IL-6 measurement by MSD, R&D, and ULX but not LX, and the superiority of MSD with respect to dynamic range. Plasma IL-6 concentrations increase in response to glucose and insulin, consistent with both an early glucose-dependent response (detectable at 1-2 hours and a late insulin-dependent response

  12. Targeted deposition of antibodies on a multiplex CMOS microarray and optimization of a sensitive immunoassay using electrochemical detection.

    Directory of Open Access Journals (Sweden)

    John Cooper

    2010-03-01

    Full Text Available The CombiMatrix ElectraSense microarray is a highly multiplex, complementary metal oxide semiconductor with 12,544 electrodes that are individually addressable. This platform is commercially available as a custom DNA microarray; and, in this configuration, it has also been used to tether antibodies (Abs specifically on electrodes using complementary DNA sequences conjugated to the Abs.An empirical method is described for developing and optimizing immunoassays on the CombiMatrix ElectraSense microarray based upon targeted deposition of polypyrrole (Ppy and capture Ab. This process was automated using instrumentation that can selectively apply a potential or current to individual electrodes and also measure current generated at the electrodes by an enzyme-enhanced electrochemical (ECD reaction. By designating groups of electrodes on the array for different Ppy deposition conditions, we determined that the sensitivity and specificity of a sandwich immunoassay for staphylococcal enterotoxin B (SEB is influenced by the application of different voltages or currents and the application time. The sandwich immunoassay used a capture Ab adsorbed to the Ppy and a reporter Ab labeled for fluorescence detection or ECD, and results from these methods of detection were different.Using Ppy deposition conditions for optimum results, the lower limit of detection for SEB using the ECD assay was between 0.003 and 0.01 pg/ml, which represents an order of magnitude improvement over a conventional enzyme-linked immunosorbant assay. In the absence of understanding the variables and complexities that affect assay performance, this highly multiplexed electrode array provided a rapid, high throughput, and empirical approach for developing a sensitive immunoassay.

  13. EIA scoping in England and Wales: Practitioner approaches, perspectives and constraints

    International Nuclear Information System (INIS)

    Wood, Graham; Glasson, John; Becker, Julia

    2006-01-01

    Scoping plays a critical role in shaping the nature and extent of the environmental information subsequently contained in an Environmental Impact Statement (EIS), yet it has been identified as an area of weakness in practice () and remains a poorly understood and under-researched component of EIA. This paper provides a detailed empirical investigation and assessment of recent scoping activity in England and Wales, in the context of evolving European Union (EU) regulations. Conceptual and contextual issues associated with scoping are outlined prior to the presentation of key findings, including: approaches to scoping; the assessment of significance; characteristics and influence of consultation; and perceived constraints to scoping. The paper concludes with an overview of findings, possible explanatory factors, and recommendations for future practice

  14. NGOs Participation in the Swedish EIA Process to Establish a Nuclear Waste Disposal

    International Nuclear Information System (INIS)

    Holmstrand, Olov

    2006-01-01

    Swedish environmental NGOs have no complete consensus on the issue of nuclear waste management. However, concerning the demands on the the EIA process most of the opinions coincide. The following standpoints generally reflect those represented by MKG as interpreted by the author Continuation of nuclear waste production, also in connection with uranium mining, is inconsistent with sustainable development. The problems of nuclear waste management must be dealt with now and not left to an undecided future. However, this does not automatically mean that any final solution needs be implemented within a short period of time. Irrespective of storage or disposal method nuclear waste is a possible source for nuclear weapons for a very long time and must therefore be subject to long-term safeguards. Any storage or disposal must be designed considering the risk of intention or unintentional intrusion. The management of nuclear waste is a national task. The thus be performed on a national scale, not as now in the municipal and to some extent regional scale. The choice of method should precede the choice of site. The choice of method should be made according to a systematic process and considering functional conditions set up in advance. Different alternatives should be evaluated and compared according to strict long-term environmental standards that comply with sustainable development. This demands extensive information on more than one possible method. The choice of site should also be made according to a systematic process considering functional conditions set up in advance. A clear and understandable sieving process at a national scale should be performed to find the best possible site considering environmental conditions. Changes have to be made so that an independent body supervises the EIA process instead of the nuclear industry. This increases the chance that the choice of method and site gain legitimacy and acceptance in the eyes of ordinary citizens

  15. Road-corridor planning in the EIA procedure in Spain. A review of case studies

    International Nuclear Information System (INIS)

    Loro, Manuel; Arce, Rosa M.; Ortega, Emilio

    2014-01-01

    The assessment of different alternatives in road-corridor planning must be based on a number of well-defined territorial variables that serve as decision making criteria, and this requires a high-quality preliminary environmental assessment study. In Spain the formal specifications for the technical requirements stipulate the constraints that must be considered in the early stages of defining road corridors, but not how they should be analyzed and ranked. As part of the feasibility study of a new road definition, the most common methodology is to establish different levels of Territorial Carrying Capacity (TCC) in the study area in order to summarize the territorial variables on thematic maps and to ease the tracing process of road-corridor layout alternatives. This paper explores the variables used in 22 road-construction projects conducted by the Ministry of Public Works that were subject to the Spanish EIA regulation and published between 2006 and 2008. The aim was to evaluate the quality of the methods applied and the homogeneity and suitability of the variables used for defining the TCC. The variables were clustered into physical, environmental, land-use and cultural constraints for the purpose of comparing the TCC values assigned in the studies reviewed. We found the average quality of the studies to be generally acceptable in terms of the justification of the methodology, the weighting and classification of the variables, and the creation of a synthesis map. Nevertheless, the methods for assessing the TCC are not sufficiently standardized; there is a lack of uniformity in the cartographic information sources and methodologies for the TCC valuation. -- Highlights: • We explore 22 road-corridor planning studies subjected to the Spanish EIA regulation. • We analyze the variables selected for defining territorial carrying capacity. • The quality of the studies is acceptable (methodology, variable weighting, mapping). • There is heterogeneity in the

  16. Road-corridor planning in the EIA procedure in Spain. A review of case studies

    Energy Technology Data Exchange (ETDEWEB)

    Loro, Manuel, E-mail: manuel.loro@upm.es [Department of Urban and Regional Planning and Environment, Civil Engineering School, Universidad Politécnica de Madrid, Prof. Aranguren s/n, 28040 Madrid (Spain); Transport Research Centre (TRANSyT-UPM) Universidad Politécnica de Madrid, ETSI Caminos, Canales y Puertos, Prof. Aranguren s/n, 28040 Madrid (Spain); Centro de investigación del transporte, TRANSyT-UPM, ETSI Caminos, Canales y Puertos, Universidad Politécnica de Madrid, Prof. Aranguren s/n, 28040 Madrid (Spain); Arce, Rosa M., E-mail: rosa.arce.ruiz@upm.es [Department of Urban and Regional Planning and Environment, Civil Engineering School, Universidad Politécnica de Madrid, Prof. Aranguren s/n, 28040 Madrid (Spain); Transport Research Centre (TRANSyT-UPM) Universidad Politécnica de Madrid, ETSI Caminos, Canales y Puertos, Prof. Aranguren s/n, 28040 Madrid (Spain); Centro de investigación del transporte, TRANSyT-UPM, ETSI Caminos, Canales y Puertos, Universidad Politécnica de Madrid, Prof. Aranguren s/n, 28040 Madrid (Spain); Ortega, Emilio, E-mail: e.ortega@upm.es [Transport Research Centre (TRANSyT-UPM) Universidad Politécnica de Madrid, ETSI Caminos, Canales y Puertos, Prof. Aranguren s/n, 28040 Madrid (Spain); Centro de investigación del transporte, TRANSyT-UPM, ETSI Caminos, Canales y Puertos, Universidad Politécnica de Madrid, Prof. Aranguren s/n, 28040 Madrid (Spain); Department of Construction and Rural Roads, Forestry Engineering School, Universidad Politécnica de Madrid, Ciudad Universitaria s/n, 28040 Madrid (Spain); and others

    2014-01-15

    The assessment of different alternatives in road-corridor planning must be based on a number of well-defined territorial variables that serve as decision making criteria, and this requires a high-quality preliminary environmental assessment study. In Spain the formal specifications for the technical requirements stipulate the constraints that must be considered in the early stages of defining road corridors, but not how they should be analyzed and ranked. As part of the feasibility study of a new road definition, the most common methodology is to establish different levels of Territorial Carrying Capacity (TCC) in the study area in order to summarize the territorial variables on thematic maps and to ease the tracing process of road-corridor layout alternatives. This paper explores the variables used in 22 road-construction projects conducted by the Ministry of Public Works that were subject to the Spanish EIA regulation and published between 2006 and 2008. The aim was to evaluate the quality of the methods applied and the homogeneity and suitability of the variables used for defining the TCC. The variables were clustered into physical, environmental, land-use and cultural constraints for the purpose of comparing the TCC values assigned in the studies reviewed. We found the average quality of the studies to be generally acceptable in terms of the justification of the methodology, the weighting and classification of the variables, and the creation of a synthesis map. Nevertheless, the methods for assessing the TCC are not sufficiently standardized; there is a lack of uniformity in the cartographic information sources and methodologies for the TCC valuation. -- Highlights: • We explore 22 road-corridor planning studies subjected to the Spanish EIA regulation. • We analyze the variables selected for defining territorial carrying capacity. • The quality of the studies is acceptable (methodology, variable weighting, mapping). • There is heterogeneity in the

  17. Hapten synthesis and antibody production for the development of a melamine immunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Lei Hongtao; Shen Yudong; Song Lijun; Yang Jinyi [Key Laboratory of Food Safety of Guangdong Province/Institute of Food Quality and Safety, South China Agricultural University, Wushan Road, Tianhe District, Guangzhou 510642, Guangdong (China); Chevallier, Olivier P.; Haughey, Simon A. [Institute of Agri-Food and Land Use, Queen' s University Belfast, Belfast BT9 5AG, Northern Ireland (United Kingdom); Wang Hong [Key Laboratory of Food Safety of Guangdong Province/Institute of Food Quality and Safety, South China Agricultural University, Wushan Road, Tianhe District, Guangzhou 510642, Guangdong (China); Sun Yuanming, E-mail: ymsun@scau.edu.cn [Key Laboratory of Food Safety of Guangdong Province/Institute of Food Quality and Safety, South China Agricultural University, Wushan Road, Tianhe District, Guangzhou 510642, Guangdong (China); Elliott, Christopher T., E-mail: chris.elliott@qub.ac.uk [Institute of Agri-Food and Land Use, Queen' s University Belfast, Belfast BT9 5AG, Northern Ireland (United Kingdom)

    2010-04-14

    The incorporation of melamine into food products is banned but its misuse has been widely reported in both animal feeds and food. The development of a rapid screening immunoassay for monitoring of the substance is an urgent requirement. Two haptens of melamine were synthesized by introducing spacer arms of different lengths and structures on the triazine ring of the analyte molecular structure. 6-Aminocaproic acid and 3-mercaptopropionic acid were reacted with 2-chloro-4,6-diamino-1,3,5-triazine (CAAT) to produce hapten 1 [3-(4,6-diamino-1,6-dihydro-1,3,5-triazin-2-ylamino) hexanoic acid] and hapten 2 [3-(4,6-diamino-1,6-dihydro-1,3,5-triazin-2-ylthio) propanoic acid], respectively. The molecular structures of the two haptens were identified by {sup 1}H nuclear magnetic resonance spectrometry, mass spectrometry and infrared spectrometry. An immunogen was prepared by coupling hapten 1 to bovine serum albumin (BSA). Two plate coating antigens were prepared by coupling both haptens to egg ovalbumin (OVA). A competitive indirect enzyme-linked immunosorbent assay (ciELISA) was developed to evaluate homogeneous and heterogeneous assay formats. The results showed that polyclonal antibodies with high titers were obtained, and the heterogeneous immunoassay format demonstrated a better performance with an IC{sub 50} of 70.6 ng mL{sup -1}, a LOD of 2.6 ng mL{sup -1} and a LOQ of 7.6 ng mL{sup -1}. Except for cyromazine, no obvious cross-reactivity to common compounds was found. The data showed that the hapten synthesis was successful and the resultant antisera could be used in an immunoassay for the rapid and sensitive detection of this banned chemical.

  18. A multiscale method for assessing vegetation baseline of Environmental Impact Assessment (EIA) in protected areas of Chile

    Science.gov (United States)

    Anibal Pauchard; Eduardo Ugarte; Jaime Millan

    2000-01-01

    The exponential growth of recreation and tourism or ecotourism activities is affecting ecological processes in protected areas of Chile. In order to protect protected areas integrity, all projects inside their boundaries must pass through the Environmental Impact Assessment (EIA). The purpose of this research was to design a multiscale method to assess vegetation for...

  19. Enzyme assays.

    Science.gov (United States)

    Brodelius, P E

    1991-02-01

    The past year or so has seen the development of new enzyme assays, as well as the improvement of existing ones. Assays are becoming more rapid and sensitive as a result of modifications such as amplification of the enzyme product(s). Recombinant DNA technology is now being recognized as a particularly useful tool in the search for improved assay systems.

  20. Seismo-Ionospheric Coupling as Intensified EIA Observed by Satellite Electron Density and GPS-TEC Data

    Science.gov (United States)

    Ryu, K.; Jangsoo, C.; Kim, S. G.; Jeong, K. S.; Parrot, M.; Pulinets, S. A.; Oyama, K. I.

    2014-12-01

    Examples of intensified EIA features temporally and spatially related to large earthquakes observed by satellites and GPS-TEC are introduced. The precursory, concurrent, and ex-post enhancements of EIA represented by the equatorial electron density, which are thought to be related to the M8.7 Northern Sumatra earthquake of March 2005, the M8.0 Pisco earthquake of August 2007, and the M7.9 Wenchuan Earthquake of 12 May 2008, are shown with space weather condition. Based on the case studies, statistical analysis on the ionospheric electron density data measured by the Detection of Electro-Magnetic Emissions Transmitted from Earthquake Regions satellite (DEMETER) over a period of 2005-2010 was executed in order to investigate the correlation between seismic activity and equatorial plasma density variations. To simplify the analysis, three equatorial regions with frequent earthquakes were selected and then one-dimensional time series analysis between the daily seismic activity indices and the EIA intensity indices were performed for each region with excluding the possible effects from the geomagnetic and solar activity. The statistically significant values of the lagged cross-correlation function, particularly in the region with minimal effects of longitudinal asymmetry, indicate that some of the very large earthquakes with M > 7.0 in the low latitude region can accompany observable seismo-ionospheric coupling phenomena in the form of EIA enhancements, even though the seismic activity is not the most significant driver of the equatorial ionospheric evolution. The physical mechanisms of the seismo-ionospheric coupling to explain the observation and the possibility of earthquake prediction using the EIA intensity variation are discussed.

  1. Comparison of immunoassays for differentiation of herpes simplex virus type 1 and 2 antibodies

    International Nuclear Information System (INIS)

    Klapper, Paul E.; Valley, Pam J.; Cleator, Gerham M.; Mandall, D.; Qutub, Mohammed O.

    2006-01-01

    To asses the commercial available enzyme-linked immunosorbent assays (ELISA) for differentiation of herpes simplex virus type 1 (Hs-1) and type 2 (HSV-2) antibodies. The study was performed between January 1997 to November 2002 in the Division ofVirology,Department of Pathological Sciences, Central Manchester Healthcare Trust and University of Manchester, Manchester, United Kingdom. Assays based upon type-specific glycoprotein G-1 (gG-1) for HSV-1, and glycoprotein G-2 (gG-2) from HSV-2 were evaluated to differentiate between HSV-1 and HSV-2 antibodies. Using 5 different ELISA tests, 2 panels of serum samples were tested. Panel one consisted of 88 sera, selected from the serum bank of the Clinical Virology Laboratory, Manchester Royal Infirmary; panel 2 comprised of 90 sera selected from samples collected from Bangladeshi female commercial workers.The data of this study showed that a high rate of gG-1 based immunoassays ranged from 87.9-100% for sensitivity and 51.5-100% specificity. Although there are several immunoassays were claimed to differentiate between HSV-1 and HSV-2 antibodies, selection of these assays should be carefully interpreted with the overall clinical framework provided by detailed sexual history and genital examination. (author)

  2. Production of monoclonal antibody against clonazepam for immunoassay of benzodiazepine drugs in swine tissues.

    Science.gov (United States)

    Shan, Wen C; Cui, Ya L; He, Xin; Zhang, Lei; Liu, Jing; Wang, Jian P

    2015-01-01

    The objective of the present study was to produce a generic monoclonal antibody for immunoassay of residues of benzodiazepine drugs in swine tissues. Clonazepam was used to synthesize a hapten that was coupled to bovine serum albumin as an immunogen for the production of monoclonal antibody. Results showed that the obtained monoclonal antibody was able to recognize five benzodiazepine drugs simultaneously (clonazepam, flunitrazepam nitrazepam, diazepam, and oxazepam). The cross-reactivities were in the range of 24-100% and the limits of detection were in the range of 0.2-1.5 ng mL(-1) depending on the drug. Then a competitive indirect enzyme-linked immunosorbent assay was developed to determine the residues of five benzodiazepines in swine tissues (muscle, liver and kidney). The recoveries of five analytes from the fortified blank samples were in the range of 74.5-96.5% with coefficients of variation lower than 16.7%. Therefore, this immunoassay could be used as a rapid and simple method for the screening of residues of five benzodiazepine drugs in animal-derived foods.

  3. Alkaline phosphatase labeled SERS active sandwich immunoassay for detection of Escherichia coli

    Science.gov (United States)

    Bozkurt, Akif Goktug; Buyukgoz, Guluzar Gorkem; Soforoglu, Mehmet; Tamer, Ugur; Suludere, Zekiye; Boyaci, Ismail Hakki

    2018-04-01

    In this study, a sandwich immunoassay method utilizing enzymatic activity of alkaline phosphatase (ALP) on 5-bromo-4-chloro-3-indolyl phosphate (BCIP) for Escherichia coli (E. coli) detection was developed using surface enhanced Raman spectroscopy (SERS). For this purpose, spherical magnetic gold coated core-shell nanoparticles (MNPs-Au) and rod shape gold nanoparticles (Au-NRs) were synthesized and modified for immunomagnetic separation (IMS) of E. coli from the solution. In order to specify the developed method to ALP activity, Au-NRs were labeled with this enzyme. After successful construction of the immunoassay, BCIP substrate was added to produce the SERS-active product; 5-bromo-4-chloro-3-indole (BCI). A good linearity (R2 = 0.992) was established between the specific SERS intensity of BCI at 600 cm- 1 and logarithmic E. coli concentration in the range of 1.7 × 101-1.7 × 106 cfu mL- 1. LOD and LOQ values were also calculated and found to be 10 cfu mL- 1 and 30 cfu mL- 1, respectively.

  4. A summary report on the 23rd quality control survey for immunoassays in Japan, 2001

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2002-10-01

    The purpose of the survey is to improve the quality of in vitro tests and this report is its summary of immunoassays (the old name: RI in vitro tests) conducted in 2001. The survey was performed in 143 facilities out of 1,655 in Japan, which involved 20 national and public university hospitals, 16 private university hospitals, 21 national and public hospitals, 26 private hospitals, 41 hygiene test institutes and 19 reagent manufacturers. Tests examined were on 6 substances related to functions of pituitary, 5 of thyroid, 1 of parathyroid, 4 of gastro-intestine and pancreas, 5 of gonad and placenta, 4 of adrenal, 1 of renal-blood pressure regulation, on IgE, on digoxin and on 12 tumor-related substances. Tests were done on 2-3 samples supplied from the Committee and the mean, standard deviation and coefficient of variation were calculated for one way analysis of variance of within- and between-kit. Methods included those (65.4% vs 62.7% in 2000) with non-radioisotope like enzyme immunoassay (non-RI) and with radioisotopes like radioimmunoassay (RI). Dissociation between manufacturers of non-RI values without a common standard substance and between non-RI and RI values even with the same antibody was noted: the Committee considered these problems as their future task. (K.H.)

  5. Strengthening research on animal reproduction and disease diagnosis in Asia through the application of immunoassay techniques

    International Nuclear Information System (INIS)

    1994-02-01

    This publication contains the results presented by participants of a final Research Co-ordination Meeting which was held from 1 to 5 February 1993 at the University of Chulalongkorn, Bangkok, Thailand, as part of an FAO/IAEA Co-ordinated Research Programme on Strengthening Research on Animal Reproduction and Disease Diagnosis in Asia through the Application of Immunoassay Techniques. The purpose of this Programme was essentially to encourage national livestock production and veterinary institutes in Asia to conduct on-farm research into existing constraints on animal productivity and ways of reducing or removing these through low cost changes in management. Emphasis was given to defining existing levels of reproductive efficiency in indigeneous livestock and examining responses to nutritional or other interventions, and to exploring possibilities for using new approaches for diagnosing and controlling some diseases considered to impact adversely on Asian livestock production. Within the framework of all studies, immunoassay (radioimmunoassay and enzyme-linked immunosorbent assay) were employed for measuring levels of reproductive hormones or detecting antibodies to a variety of disease causing agents, the aim being to gain better insight into the underlying nature of the problems being encountered and reasons for the success or otherwise of approaches taken for their resolution. Refs, figs and tabs

  6. Protein immunoassay methods for detection of biotech crops: applications, limitations, and practical considerations.

    Science.gov (United States)

    Stave, James W

    2002-01-01

    Immunoassay methods are available for detection and quantitation of proteins expressed by most biotechnology-derived crops in commercial production. The 2 most common test formats are enzyme-linked immunosorbent assay (ELISA) and immunochromatographic (lateral flow) strip tests. Two ELISA methods, one for Roundup Ready soybeans and one for MON810 CrylAb corn, were the subject of large international collaborative studies and were demonstrated to quantitatively determine the concentrations of biotech crops in samples of ground grain. Quantitative ELISA methods are also useful for analysis of processed fractions of agricultural commodities such as soybean toasted meal or corn flour. Both strip tests and ELISAs for biotech crops are currently being used on a large scale in the United States to manage the sale and distribution of grain. In these applications, tests are used to determine if the concentration of biotech grain is above or below specified threshold limits. Using existing U.S. Department of Agriculture sampling techniques, the reliability of the threshold determination is expressed in terms of statistical confidence rather than analytical precision. Combining the use of protein immunoassays with Identity Preservation systems provides an effective means of characterizing the raw and processed agricultural inputs to the food production system in a way that allows food producers to comply with labeling laws.

  7. Multicenter Analytical Validation of Aβ40 Immunoassays

    Directory of Open Access Journals (Sweden)

    Linda J. C. van Waalwijk van Doorn

    2017-07-01

    Full Text Available BackgroundBefore implementation in clinical practice, biomarker assays need to be thoroughly analytically validated. There is currently a strong interest in implementation of the ratio of amyloid-β peptide 1-42 and 1-40 (Aβ42/Aβ40 in clinical routine. Therefore, in this study, we compared the analytical performance of six assays detecting Aβ40 in cerebrospinal fluid (CSF in six laboratories according to a recently standard operating procedure (SOP developed for implementation of ELISA assays for clinical routine.MethodsAβ40 assays of six vendors were validated in up to three centers per assay according to recently proposed international consensus validation protocols. The performance parameters included sensitivity, precision, dilutional linearity, recovery, and parallelism. Inter-laboratory variation was determined using a set of 20 CSF samples. In addition, test results were used to critically evaluate the SOPs that were used to validate the assays.ResultsMost performance parameters of the different Aβ40 assays were similar between labs and within the predefined acceptance criteria. The only exceptions were the out-of-range results of recovery for the majority of experiments and of parallelism by three laboratories. Additionally, experiments to define the dilutional linearity and hook-effect were not executed correctly in part of the centers. The inter-laboratory variation showed acceptable low levels for all assays. Absolute concentrations measured by the assays varied by a factor up to 4.7 for the extremes.ConclusionAll validated Aβ40 assays appeared to be of good technical quality and performed generally well according to predefined criteria. A novel version of the validation SOP is developed based on these findings, to further facilitate implementation of novel immunoassays in clinical practice.

  8. Immunoassay and polymerase chain reaction techniques for ...

    African Journals Online (AJOL)

    Objectives: To compare the Reverse Passive Latex Agglutination (RPLA) and Enzyme Linked Immunosorbent Assay (ELISA) techniques with a Polymerase Chain Reaction (PCR) for detection of enterotoxigenic Bacillus cereus. Design: A cross-sectional study. Setting: The Department of Public Health, Pharmacology and ...

  9. Detection of 3-phenoxybenzoic acid in river water with a colloidal gold-based lateral flow immunoassay.

    Science.gov (United States)

    Liu, Yuan; Wu, Aihua; Hu, Jing; Lin, Manman; Wen, Mengtang; Zhang, Xiao; Xu, Chongxin; Hu, Xiaodan; Zhong, Jianfeng; Jiao, Lingxia; Xie, Yajing; Zhang, Cunzhen; Yu, Xiangyang; Liang, Ying; Liu, Xianjin

    2015-08-15

    3-Phenoxybenzoic acid (3-PBA) is a general metabolite of synthetic pyrethroids. It could be used as a generic biomarker for multiple pyrethroids exposure for human or pyrethroid residues in the environment. In this study, monoclonal antibodies (mAbs) against 3-PBA were developed by using PBA-bovine serum albumin (BSA) as an immunogen. In the competitive enzyme-linked immunosorbent assay (ELISA) format, the I50 and I10 values of purified mAbs were 0.63 and 0.13 μg/ml, respectively, with a dynamic range between 0.19 and 2.04 μg/ml. Then, the colloidal gold (CG)-based lateral flow immunoassay was established based on the mAbs. The working concentration of coating antigen and CG-labeled antibodies and the blocking effects were investigated to get optimal assay performance. The cutoff value for the assay was 1 μg/ml 3-PBA, and the detection time was within 10 min. A total of 40 river water samples were spiked with 3-PBA at different levels and determined by the lateral flow immunoassay without any sample pretreatments. The negative false rate was 2.5%, and no positive false results were observed at these levels. This lateral flow immunoassay has the potential to be an on-site screening method for monitoring 3-PBA or pyrethroid residues in environmental samples. Copyright © 2015 Elsevier Inc. All rights reserved.

  10. Evaluation of the double diffusion, enzyme immunoassay and immunoblotting techniques, for the diagnosis of human hydatid disease in tropical areas Evaluación de las técnicas de Doble Difusión, Ensayo Inmunoenzimático e Inmunoblotting en el diagnóstico de la hidatidosis humana en áreas tropicales

    Directory of Open Access Journals (Sweden)

    Sandra Planchart

    1994-06-01

    Full Text Available Hydatid disease in tropical areas poses a serious diagnostic problem due to the high frequence of cross-reactivity with other endemic helminthic infections. The enzyme-linked-immunosorbent assay (ELISA and the double diffusion arc 5 showed respectively a sensitivity of 73% and 57% and a specificity of 84-95% and 100%. However, the specificity of ELISA was greatly increased by using ovine serum and phosphorylcholine in the diluent buffer. The hydatic antigen obtained from ovine cyst fluid showed three main protein bands of 64,58 and 30 KDa using SDS PAGE and immunoblotting. Sera from patients with onchocerciasis, cysticercosis, toxocariasis and Strongyloides infection cross-reacted with the 64 and 58 KDa bands by immunoblotting. However, none of the analyzed sera recognized the 30 KDa band, that seems to be specific in this assay. The immunoblotting showed a sensitivity of 80% and a specificity of 100% when used to recognize the 30 KDa band.La Hidatidosis en áreas tropicales representa un serio problema diagnóstico por la alta frecuencia de reactividad cruzada con otras infecciones helmínticas. Las técnicas de inmunoensayo enzimático (ELISA y doble difusión arco 5 (DD5 mostraron una sensibilidad de 73 y 57 % y una especificidad de 84-95% y 100%, respectivamente. La especificidad en la técnica de ELISA, fue mejorada sustancialmente al emplear como diluyente de los sueros una solución buffer conteniendo suero ovino normal y fosforilcolina. En líquido obtenido de hidátides de Echinococcus granulosus de origen ovino, se demostraron tres bandas de origen proteico de 64, 58 y 30 KDa de peso molecular, empleando SDS e inmunoblotting. Sueros de pacientes con estrongiloidiasis, oncocer-cosis, toxocariasis y cisticercosis reaccionaron con las bandas de 64 y 58 KDa en inmunoblotting. Sin embargo ninguno de los sueros reconoció la banda de 30 KD, la cual parece ser específica para este ensayo. La técnica de inmunobloting empleada para poner en

  11. Evaluation of an enzyme immunoassay for clinical diagnosis of neurocysticercosis in symptomatic patients Avaliação de um teste imunoenzimático para o diagnóstico clínico de neurocisticercose em pacientes sintomáticos

    Directory of Open Access Journals (Sweden)

    Reynaldo Mendes de Carvalho Junior

    2010-12-01

    Full Text Available INTRODUCTION: Neurocysticercosis is an infection of the human central nervous system caused by the metacestode larvae of Taenia solium. Neurocysticercosis is the most common parasitic disease in developing countries. Epilepsy is the most common clinical manifestation. Difficulties in confirming the diagnosis motivated the evaluation of the enzyme-linked immunosorbent assay on cerebral spinal fluid (CSF. METHODS: Twenty-two patients with NCC and 44 control patients were studied. CSF was analyzed using a commercial ELISA kit developed for NCC. Sensitivity and specificity were measured and a multivariate logistic regression was performed. RESULTS: Sensitivity and specificity of ELISA were 31.8% and 100%, respectively, with accuracy of 77.3%. Only the size of the lesions proved to be important for performance of the test. CONCLUSIONS: The results showed that ELISA contributes to the diagnosis of neurocysticercosis if the result is negative or if the patient has a lesion of 2 cm or more.INTRODUÇÃO: Neurocisticercose é a infecção do sistema nervoso central causada pela larva metacestódea da Taenia solium. Neurocisticercose é a parasitose mais comum nos países em desenvolvimento. Epilepsia é a sua manifestação clínica mais comum. A dificuldade para confirmar o diagnóstico motivou a avaliação do ensaio imunoenzimático ligado à enzima no líquido cérebro-espinhal. MÉTODOS: Vinte e dois pacientes com NCC e 44 pacientes controles foram estudados. Líquido cérebro-espinhal foi analisado por um kit ELISA comercial desenvolvido para NCC. A sensibilidade e especificidade foram medidas e uma análise multivariada de regressão logística foi realizada. RESULTADOS: A sensibilidade e a especificidade de ELISA foram, respectivamente, 31,8% e 100%, com acurácia de 77,3%. Apenas o tamanho das lesões mostrou-se importante para o desempenho do teste. CONCLUSÕES: Este estudo concluiu que ELISA contribui para o diagnóstico de NCC, caso o teste

  12. Antigen excess in modern immunoassays: to anticipate on the unexpected.

    Science.gov (United States)

    Jacobs, Joannes F M; van der Molen, Renate G; Bossuyt, Xavier; Damoiseaux, Jan

    2015-02-01

    Immunoassays measuring sera with high analyte concentration may be prone to an artifact that causes underestimation of the analyte concentration. This phenomenon is generally described as antigen excess or the prozone effect. Characteristically, serum with high concentrations of a certain analyte can give a false negative/low result when tested at the recommended dilution, but reacts strongly positive upon further dilution. Increased insight of the antigen excess mechanisms and tools to prevent it has reduced the analytical problems caused by prozone effects in daily laboratory practice. However, misinterpretation of laboratory results caused by antigen excess does still occur, in virtually any type of immunoassay. Awareness by the laboratory specialist of the mechanisms underlying antigen excess in the different immunoassays, strategies to detect it, and adequate communication with clinicians can help to avoid reporting false negative test-results. Copyright © 2014 Elsevier B.V. All rights reserved.

  13. Synthesis-based approach toward direct sandwich immunoassay for ciguatoxin CTX3C.

    Science.gov (United States)

    Oguri, Hiroki; Hirama, Masahiro; Tsumuraya, Takeshi; Fujii, Ikuo; Maruyama, Megumi; Uehara, Hisatoshi; Nagumo, Yoko

    2003-06-25

    Ciguatoxins are the major causative toxins of ciguatera seafood poisoning. Limited availability of ciguatoxins has hampered the development of a reliable and specific immunoassay for detecting these toxins in contaminated fish. Monoclonal antibodies (mAbs) specific against both ends of ciguatoxin CTX3C were prepared by immunization of mice with protein conjugates of rationally designed synthetic haptens, 3 and 4, in place of the natural toxin. Haptenic groups that possess a surface area larger than 400 A(2) were required to produce mAbs that can bind strongly to CTX3C itself. A direct sandwich enzyme-linked immunosorbent assay (ELISA) using these mAbs was established to detect CTX3C at the ppb level with no cross-reactivity against other related marine toxins, including brevetoxin A, brevetoxin B, okadaic acid, or maitotoxin.

  14. Vertical microreactor stack consist of poly-(tetrafluoroethylene) microfluidics for immunoassay

    International Nuclear Information System (INIS)

    Ukita, Yoshiaki; Kondo, Saki; Utsumi, Yuichi; Takeo, Masahiro; Negoro, Seiji; Kataoka, Chiwa

    2010-01-01

    This paper reports the first application of high-aspect ratio PTFE microstruscute, which fabricated by synchrotron radiation induced photo-evaporation process, to enzyme-linked immunosorvent assay. The advantages of PTFE microstructure for the development of lab-on-a-chip due to the extremely high-aspect ratio microstructure and chemical stability of PTFE is discussed. The results of immunoassay shows the successful detection of analyte (mouse IgG) with detection range with 0-100ng/ml. This result suggests the successful immobilization of antibody (anti-mouse IgG goat antibody) onto the x-ray exposed surface of PTFE microstructure and successful demonstration of antigen-antibody reaction in the PTFE high-aspect ratio microstructure. We also demonstrated the detection of polychlorinated biphenyl (PCB). As the result of demonstration, we successfully detected PCB with ranging analyte concentration of 0.1-10 ng/ml. (author)

  15. Peptide dot immunoassay and immunoblotting: electroblotting from aluminum thin-layer chromatography plates and isoelectric focusing gels to activated nitrocellulose

    DEFF Research Database (Denmark)

    Bjerrum, O.J.; Holm, A.; Lauritzen, Edgar

    1993-01-01

    Peptide dot immunoassay, electroblotting, activated nitrocellulose, dot blot, membranes, peptides and proteins......Peptide dot immunoassay, electroblotting, activated nitrocellulose, dot blot, membranes, peptides and proteins...

  16. NiCoBP-doped carbon nanotube hybrid: A novel oxidase mimetic system for highly efficient electrochemical immunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Bing; He, Yu; Liu, Bingqian; Tang, Dianping, E-mail: dianping.tang@fzu.edu.cn

    2014-12-03

    Highlights: • We report a new oxidase mimetic system for highly efficient electrochemical immunoassay. • NiCoBP-doped carbon nanotube hybrids were used as the nanocatalysts. • NiCoBP-doped carbon nanotube hybrids were used as the mimic oxidase. - Abstract: NiCoBP-doped multi-walled carbon nanotube (NiCoBP–MWCNT) was first synthesized by using induced electroless-plating method and functionalized with the biomolecules for highly efficient electrochemical immunoassay of prostate-specific antigen (PSA, used as a model analyte). We discovered that the as-synthesized NiCoBP–MWCNT had the ability to catalyze the glucose oxidization with a stable and well-defined redox peak. The catalytic current increased with the increment of the immobilized NiCoBP–MWCNT on the electrode. Transmission electron microscope (TEM) and energy dispersive X-ray spectrometry (EDX) were employed to characterize the as-prepared NiCoBP–MWCNT. Using the NiCoBP–MWCNT-conjugated anti-PSA antibody as the signal-transduction tag, a new enzyme-free electrochemical immunoassay protocol could be designed for the detection of target PSA on the capture antibody-functionalized immunosensing interface. Experimental results revealed that the designed immunoassay system could exhibit good electrochemical responses toward target PSA, and allowed the detection of PSA at a concentration as low as 0.035 ng mL{sup −1}. More importantly, the NiCoBP-MWCNT-based oxidase mimetic system could be further extended for the monitoring of other low-abundance proteins or disease-related biomarkers by tuning the target antibody.

  17. PRNP variants in goats reduce sensitivity of detection of PrPSc by immunoassay

    Science.gov (United States)

    Immunoassays are extensively utilized in disease diagnostics with monoclonal antibodies serving as critical tools within the assay. Detection of scrapie in sheep and goats relies heavily on immunoassays including immunohistochemistry, western blotting, and ELISA. In the United States, regulatory tes...

  18. Distribution of Phytophthora spp. in Field Soils Determined by Immunoassay.

    Science.gov (United States)

    Miller, S A; Madden, L V; Schmitthenner, A F

    1997-01-01

    ABSTRACT Populations of Phytophthora spp. were determined by enzyme-linked immunosorbent assay (ELISA) in field soils used for pepper and soybean production in Ohio. Soybean fields were sampled extensively (64 fields, n = 6 samples per field over 2 years) and intensively (4 fields, n = 64 samples per field in 1 year) to assess heterogeneity of P. sojae populations. Four pepper fields (n = 64), three of which had a history of Phytophthora blight (caused by P. capsici), also were sampled intensively during a 6-month period. Mean (m), variance (v), and measures of aggregation (e.g., variance-to-mean ratio [v/m]) of immunoassay values, translated to Phytophthora antigen units (PAU), were related to the disease history in each of the pepper and soybean fields. Mean PAU values for fields in which Phytophthora root rot (soybean) or blight (pepper) had been moderate to severe were higher than in fields in which disease incidence had been low or not observed. A detection threshold value of 11.3 PAU was calculated with values for 64 samples from one pepper field, all of which tested negative for Phytophthora by bioassay and ELISA. Seven of the eight intensively sampled fields contained at least some detectable Phytophthora propagules, with the percentage of positive samples ranging from 1.6 to 73.4. Mean PAU values ranged from 1 to 84 (extensive soybean field sampling), 6 to 24 (intensive soybean field sampling), and 4 to 30 (intensive pepper field sampling); however, variances ranged from 0 to 7,774 (extensive sampling), 30 to 848 (intensive soybean field sampling), and 5 to 2,401 (intensive pepper field sampling). Heterogeneity of PAU was high in most individual soybean and pepper fields, with values of v/m greater than 1, and log(v) increasing with log(m), with a slope of about 2.0. Spatial autocorrelation coefficients were not significant, indicating there was no relationship of PAU values in neighboring sampling units (i.e., field locations) of the intensively sampled

  19. Comparing GIS-based habitat models for applications in EIA and SEA

    International Nuclear Information System (INIS)

    Gontier, Mikael; Moertberg, Ulla; Balfors, Berit

    2010-01-01

    Land use changes, urbanisation and infrastructure developments in particular, cause fragmentation of natural habitats and threaten biodiversity. Tools and measures must be adapted to assess and remedy the potential effects on biodiversity caused by human activities and developments. Within physical planning, environmental impact assessment (EIA) and strategic environmental assessment (SEA) play important roles in the prediction and assessment of biodiversity-related impacts from planned developments. However, adapted prediction tools to forecast and quantify potential impacts on biodiversity components are lacking. This study tested and compared four different GIS-based habitat models and assessed their relevance for applications in environmental assessment. The models were implemented in the Stockholm region in central Sweden and applied to data on the crested tit (Parus cristatus), a sedentary bird species of coniferous forest. All four models performed well and allowed the distribution of suitable habitats for the crested tit in the Stockholm region to be predicted. The models were also used to predict and quantify habitat loss for two regional development scenarios. The study highlighted the importance of model selection in impact prediction. Criteria that are relevant for the choice of model for predicting impacts on biodiversity were identified and discussed. Finally, the importance of environmental assessment for the preservation of biodiversity within the general frame of biodiversity conservation is emphasised.

  20. EIA model documentation: Petroleum market model of the national energy modeling system

    International Nuclear Information System (INIS)

    1995-01-01

    The purpose of this report is to define the objectives of the Petroleum Market Model (PMM), describe its basic approach, and provide detail on how it works. This report is intended as a reference document for model analysts, users, and the public. Documentation of the model is in accordance with EIA's legal obligation to provide adequate documentation in support of its models. The PMM models petroleum refining activities, the marketing of petroleum products to consumption regions, the production of natural gas liquids in gas processing plants, and domestic methanol production. The PMM projects petroleum product prices and sources of supply for meeting petroleum product demand. The sources of supply include crude oil, both domestic and imported; other inputs including alcohols and ethers; natural gas plant liquids production; petroleum product imports; and refinery processing gain. In addition, the PMM estimates domestic refinery capacity expansion and fuel consumption. Product prices are estimated at the Census division level and much of the refining activity information is at the Petroleum Administration for Defense (PAD) District level

  1. Assessing the performance of SIA in the EIA context: A case study of South Africa

    International Nuclear Information System (INIS)

    Pisani, Jacobus A. du; Sandham, Luke A.

    2006-01-01

    This article, a theoretical perspective based on a literature study, is a critical evaluation of SIA as part of the EIA process in South Africa against the background of international guidelines and best practices. It includes sections on the historical background of the development of SIA in South Africa, the legal status and requirements of SIA in the country, and a critical evaluation of SIA regulation in South Africa. The conclusion reached in the article is that the persistent problems of SIA practice, experienced in other parts of the world, are also evident in South Africa. Apart from institutional, financial and professional constraints, there are also serious problems associated with approach and methods. This conclusion confirms the findings of empirical studies that SIA in South Africa is neglected, that the practice of SIA in South Africa is not yet on a sound footing, and that it does not receive the professional attention it deserves in a country beset by enormous social challenges. To conclude the article recommendations are made to improve the level of SIA practice in South Africa, and the possible significance of this national case study for international practice is indicated

  2. The effects of vehicle congestion on the environment – an EIA in the Ngorongoro crater. The Environmental Impact Statement

    OpenAIRE

    Nyahongo, Julius; Lowassa, Asanterabi; Malugu, Lucas; Nkya, Hassan; Mwakalebe, Grayson; Thomassen, Jørn; Kaltenborn, Bjørn Petter; Lyamuya, Richard; Marealle, Wilfred; Keyyu, Julius; Stokke, Sigbjørn; Røskaft, Eivin

    2007-01-01

    Nyahongo, J., Lowassa, A., Malugu, L., Nkya, H., Mwakalebe, G., Thomassen, J., Kaltenborn, B. P., Lyamuya, R., Marealle, W., Keyyu, J., Stokke, S. & Røskaft, E. 2007. The effects of vehicle congestion on the environment – an EIA in the Ngorongoro crater. The Environmental Impact Statement – NINA Report 258. 83 pp. The Ngorongoro Conservation Area Authority (NCAA) faces a great challenge in managing the vehicle congestion in the Ngorongoro crater. They are in need for an assessment of the pres...

  3. Development of Indirect Competitive Immuno-Assay Method Using SPR Detection for Rapid and Highly Sensitive Measurement of Salivary Cortisol Levels

    Energy Technology Data Exchange (ETDEWEB)

    Tahara, Yusuke; Huang, Zhe; Kiritoshi, Tetsuro [Graduate School of Information Science and Electrical Engineering, Kyushu University, Fukuoka (Japan); Onodera, Takeshi [Research and Development Center for Taste and Odor Sensing, Kyushu University, Fukuoka (Japan); Toko, Kiyoshi, E-mail: toko@ed.kyushu-u.ac.jp [Graduate School of Information Science and Electrical Engineering, Kyushu University, Fukuoka (Japan); Research and Development Center for Taste and Odor Sensing, Kyushu University, Fukuoka (Japan)

    2014-05-30

    The monitoring of salivary cortisol as a key biomarker of an individual’s stress response has been increasingly focused on. This paper describes the development of a novel cortisol immuno-assay method based on an indirect competitive method using a commercially available surface plasmon resonance instrument. The surface of an Au chip was modified with PEG6-COOH aromatic dialkanethiol self-assembled monolayers and hydrocortisone 3-(O-carboxymethyl) oxime (hydrocortisone 3-CMO) as a cortisol analog. A detection limit of 38 ppt range with a measurement range of 10 ppt–100 ppb was accomplished without the incubation of a mixing solution consisting of standard cortisol and an anti-cortisol antibody, and the time for quantification of cortisol concentration was 8 min from the sample injection. We experimentally compared our immuno-assay with a commercialized salivary cortisol enzyme-linked immunosorbent assay (ELISA) kit using human saliva samples. It was found that the results obtained by the cortisol immuno-assay had a good correlation with those obtained by ELISA assay (R = 0.96). Our findings indicate the potential utility of the cortisol immuno-assay for measurements of human salivary cortisol levels.

  4. Public Participation Planning of Environmental Impact Assessment (EIA and Regulations: Analysis of Inconsistency for Some Cases in Indonesia

    Directory of Open Access Journals (Sweden)

    Hindrayani Aniek

    2018-01-01

    Full Text Available The failure in community involvement during the environmental documents planning may result in the failure of the planned project implementation. This study aims to determine the gap between practices and regulations that apply to the process of community involvement in the environmental documents planning, and find out inconsistency of implementation on each stakeholder in the planning of the Environmental Impact Assessment (EIA and the environmental permit. The method used was qualitative through interview and literature study which is analyzed using triangulation model and presented in the form of concept map. The results of the study indicate that 1 the determination of community representatives based on the criteria of the impacted communities is not clearly described, 3 suggestions, opinions, and responses to the environmental impact management are not well implemented by the project proponent, 3 implementation of the environmental management of other licensed activities affecting the behavior (4 stakeholders (project proponent, EIA consultants, and EIA appraisal committee do not play their role as mandated in applicable legislation.

  5. [Serological and clinical proof of freedom from Equine Infectious Anemia (EIA) in imported and domestic horses in Switzerland].

    Science.gov (United States)

    Kaiser, A; Meier, H P; Doherr, M G; Perler, L; Zanoni, R; Gerber, V

    2009-04-01

    Since 1991, no cases of Equine Infectious Anemia (EIA) have been reported in Switzerland. Risk factors for introduction of the virus into Switzerland are still present or have even increased as frequent inapparent infections, large numbers of imported horses, (since 2003) absence of compulsory testing prior to importation, EIA cases in surrounding Europe, possible illegal importation of horses, frequent short-term stays, poor knowledge of the disease among horse owners and even veterinarians. The aim of this study was to provide evidence of freedom from EIA in imported and domestic horses in Switzerland. The serum samples from 434 horses imported since 2003 as well as from 232 domestic horses fifteen years of age or older (since older horses have naturally had a longer time of being exposed to the risk of infection) were analysed using a commercially available ELISA test. All samples were seronegative, indicating that the maximum possible prevalence that could have been missed with this sample was 0.5% (95% confidence).

  6. Public Participation Planning of Environmental Impact Assessment (EIA) and Regulations: Analysis of Inconsistency for Some Cases in Indonesia

    Science.gov (United States)

    Hindrayani, Aniek; Purwanto

    2018-02-01

    The failure in community involvement during the environmental documents planning may result in the failure of the planned project implementation. This study aims to determine the gap between practices and regulations that apply to the process of community involvement in the environmental documents planning, and find out inconsistency of implementation on each stakeholder in the planning of the Environmental Impact Assessment (EIA) and the environmental permit. The method used was qualitative through interview and literature study which is analyzed using triangulation model and presented in the form of concept map. The results of the study indicate that 1) the determination of community representatives based on the criteria of the impacted communities is not clearly described, 3) suggestions, opinions, and responses to the environmental impact management are not well implemented by the project proponent, 3) implementation of the environmental management of other licensed activities affecting the behavior (4) stakeholders (project proponent, EIA consultants, and EIA appraisal committee) do not play their role as mandated in applicable legislation.

  7. Accounting and auditing support services for the Department of Energy, Energy Information Administration, Financial Reporting System, Form EIA-28. Final report

    Energy Technology Data Exchange (ETDEWEB)

    1983-06-30

    Conceptual audits were performed for the 26 Financial Reporting System (FRS) companies. These included both analytic and mechanical security checks to evaluate the reliability of the information collected on the Form EIA-28.

  8. Nanomaterials with enzyme-like characteristics (nanozymes): next-generation artificial enzymes.

    Science.gov (United States)

    Wei, Hui; Wang, Erkang

    2013-07-21

    Over the past few decades, researchers have established artificial enzymes as highly stable and low-cost alternatives to natural enzymes in a wide range of applications. A variety of materials including cyclodextrins, metal complexes, porphyrins, polymers, dendrimers and biomolecules have been extensively explored to mimic the structures and functions of naturally occurring enzymes. Recently, some nanomaterials have been found to exhibit unexpected enzyme-like activities, and great advances have been made in this area due to the tremendous progress in nano-research and the unique characteristics of nanomaterials. To highlight the progress in the field of nanomaterial-based artificial enzymes (nanozymes), this review discusses various nanomaterials that have been explored to mimic different kinds of enzymes. We cover their kinetics, mechanisms and applications in numerous fields, from biosensing and immunoassays, to stem cell growth and pollutant removal. We also summarize several approaches to tune the activities of nanozymes. Finally, we make comparisons between nanozymes and other catalytic materials (other artificial enzymes, natural enzymes, organic catalysts and nanomaterial-based catalysts) and address the current challenges and future directions (302 references).

  9. Immunoassay Analysis of Kanamycin in Serum Using the Tobramycin Kit

    NARCIS (Netherlands)

    Dijkstra, J. A.; Voerman, A. J.; Greijdanus, B.; Touw, D. J.; Alffenaar, J. W. C.

    Kanamycin is one of the aminoglycosides used in the treatment of multidrug-resistant tuberculosis. Blood concentrations of kanamycin are predictive for the treatment efficacy and the occurrence of side effects, and dose adjustments can be needed to optimize therapy. However, an immunoassay method

  10. Toward a dry reagent immunoassay of progesterone in bovine milk

    NARCIS (Netherlands)

    Posthuma-Trumpie, G.A.

    2008-01-01

    This thesis is aimed at the development of a dry reagent immunoassay of progesterone in cow's milk. Progesterone is a steroid hormone and regulates ovulation in female mammals. The concentration of progesterone in blood and in milk is in accordance with the reproductive cycle of the individual

  11. Is the turbidimetric immunoassay of haptoglobin phenotype-dependent?

    NARCIS (Netherlands)

    Rijn, H.J.M. van; Wilt, W. van der; Stroes, J.W.; Schrijver, J.

    Comparison of the turbidimetric immunoassay of haptoglobin with a reference method (the RID technique with appropriate correction for phenotype) clearly showed the turbidimetric assay to be phenotype-dependent. Correction factors for the three main phenotypes were calculated and reference values

  12. AN EVALUATION OF FIVE COMMERCIAL IMMUNOASSAY DATA ANALYSIS SOFTWARE SYSTEMS

    Science.gov (United States)

    An evaluation of five commercial software systems used for immunoassay data analysis revealed numerous deficiencies. Often, the utility of statistical output was compromised by poor documentation. Several data sets were run through each system using a four-parameter calibration f...

  13. Multiplex biosensor immunoassays for antibiotics in the food chain

    NARCIS (Netherlands)

    Haasnoot, W.

    2009-01-01

    The use of antibiotics in food-producing animals may result in unwanted residues in food products. The main objective of the present research was to study the development and application of fast and automated multiplex surface plasmon resonance (SPR)-based biosensor immunoassays (BIAs), based on

  14. Biosensor immunoassay for flumequine in broiler serum and muscle

    NARCIS (Netherlands)

    Haasnoot, W.; Gercek, H.; Cazemier, G.; Nielen, M.W.F.

    2007-01-01

    Flumequine (Flu) is one of the fluoroquinolones most frequently applied for the treatment of broilers in The Netherlands. For the detection of residues of Flu in blood serum of broilers, a biosensor immunoassay (BIA) was developed which was fast (7.5 min per sample) and specific (no cross-reactivity

  15. Flow cytometric immunoassay for sulfonamides in raw milk

    NARCIS (Netherlands)

    Keizer, de W.; Bienenmann-Ploum, M.; Bergwerff, A.A.; Haasnoot, W.

    2008-01-01

    Sulfonamide antibiotics are applied in veterinary medicine for the treatment of microbial infections. For the detection of residues of sulfonamides in milk, a multi-sulfonamide flow cytometric immunoassay (FCI) was developed using the Luminex MultiAnalyte Profiling (xMAP) technology. In this

  16. Evaluating public participation in Chinese EIA. An integrated Public Participation Index and its application to the case of the New Beijing Airport

    International Nuclear Information System (INIS)

    Brombal, Daniele; Moriggi, Angela; Marcomini, Antonio

    2017-01-01

    In recent years, China's government authorities have devoted increasing attention to the role of public participation processes in Environmental Impact Assessment (EIA). The capacity of these processes to influence decision-making remains widely debated. This paper aims at appraising the institutional rationale informing the implementation of public participation in China's EIA, benchmarking it against three conceptualisations: (1) Normative, based on objectives of empowerment and democratisation; (2) Substantive, where participation is pursued mainly to improve quality of decisions; (3) Instrumental, seeking participation as an instrument to legitimise decision-making processes. The appraisal is carried out by means of a new integrated index (Public Participation Index, PPI), which is applied to a case study representative of latest advancements in EIA public participation practices in China, namely the “New Beijing Airport Project”. Located 46 km south of downtown Beijing, the project was approved in 2014 and it is currently under construction. Results of the PPI application to this case study indicate that, despite progress made in recent years, the implementation of public participation in Chinese EIA still largely responds to an instrumental rationale, with limited capacity for the public to affect decisions. - Highlights: • In recent years China has strengthened EIA public participation (PP) legislation. • Despite progress, implementation of PP remains informed by an instrumental rationale. • A large gap exists between principles enunciated in regulations and implementation. • The Public Participation Index can be used to monitor China's EIA PP development.

  17. Comparison of capillary electrophoresis-based immunoassay with fluorescence polarization immunoassay for the immunodetermination of methamphetamine using various methamphetamine antibodies.

    Science.gov (United States)

    Choi, J; Kim, C; Choi, M J

    1998-11-01

    An accurate and simple immunoassay using capillary electrophoresis (CE) with laser-induced fluorescence (LIF) was performed for the detection of methamphetamine (MA) in urine. The CE-LIF was conducted with an untreated fused-silica column using antiserum and a tracer of fluorescein isothiocyanate (FITC)-labeled MA. This CE-LIF system was compared with fluorescence polarization immunoassay (FPIA) in a TDx analyzer in the photo-check mode using the same FITC-labeled tracer and the same antiserum. Various antibodies, not only those prepared by our own immunogens but also those from commercial sources, were screened and characterized in both assay systems with regard to sensitivity, precision, and cross-reactivity. Both systems satisfied analytical precision and gave similar cross-reactivity patterns. However, the CE-LIF-based immunoassay was approximately one order superior to FPIA in sensitivity, requiring less volume of sample, antiserum, and tracer for the assay. Considering that the FPIA system is well known to be a useful tool for screening antibodies and detecting drugs, the CE-LIF-based immunoassay system, which is seemingly more advantageous than the FPIA system, appears to have great power for the characterization of antibodies and for the detection of MA in urine.

  18. Tyr Phosphatase-Mediated P-ERK Inhibition Suppresses Senescence in EIA + v-raf Transformed Cells, Which, Paradoxically, Are Apoptosis-Protected in a MEK-Dependent Manner

    Directory of Open Access Journals (Sweden)

    Stefania De Vitis

    2011-02-01

    Full Text Available Activation of the Ras-Raf-extracellular signal-regulated kinase (ERK pathway causes not only proliferation and suppression of apoptosis but also the antioncogenic response of senescence. How these contrasting effects are reconciled to achieve cell transformation and cancer formation is poorly understood. In a system of two-step carcinogenesis (dedifferentiated PC EIA, transformed PC EIA-polyoma-middle T [PC EIA + Py] and PC EIA-v-raf [PC EIA + raf] cells], v-raf cooperated with EIA by virtue of a strong prosurvival effect, not elicited by Py-middle T, evident toward serum-deprivation-and H2O2-induced apoptosis. Apoptosis was detected by DNA fragmentation and annexin V staining. The prosurvival function of v-raf was, in part, mitogen-activated protein kinase/ERK kinase (MEK-dependent, as shown by pharmacological MEK inhibition. The MEK-dependent antiapoptotic effect of v-raf was exerted despite a lower level of P-ERK1/2 in EIA + raf cells with respect to EIA + Py/EIA cells, which was dependent on a high tyrosine phosphatase activity, as shown by orthovanadate blockade. An ERK1/2 tyrosine phosphatase was likely involved. The high tyrosine phosphatase activity was instrumental to the complete suppression of senescence, detected by β-galactosidase activity, because tyrosine phosphatase blockade induced senescence in EIA + raf but not in EIA + Py cells. High tyrosine phosphatase activity and evasion from senescence were confirmed in an anaplastic thyroid cancer cell line. Therefore, besides EIA, EIA + raf cells suppress senescence through a new mechanism, namely, phosphatase-mediated P-ERK1/2 inhibition, but, paradoxically, retain the oncogenic effects of the Raf-ERK pathway. We propose that the survival effect of Raf is not a function of absolute P-ERK1/2 levels at a given time but is rather dynamically dependent on greater variations after an apoptotic stimulus.

  19. Effects of saliva collection using cotton swabs on melatonin enzyme immunoassay.

    Science.gov (United States)

    Kozaki, Tomoaki; Lee, Soomin; Nishimura, Takayuki; Katsuura, Tetsuo; Yasukouchi, Akira

    2011-01-10

    Although various acceptable and easy-to-use devices have been used for saliva collection, cotton swabs are among the most common ones. Previous studies reported that cotton swabs yield a lower level of melatonin detection. However, this statistical method is not adequate for detecting an agreement between cotton saliva collection and passive saliva collection, and a test for bias is needed. Furthermore, the effects of cotton swabs have not been examined at lower melatonin level, a level at which melatonin is used for assessment of circadian rhythms, namely dim light melatonin onset (DLMO). In the present study, we estimated the effect of cotton swabs on the results of salivary melatonin assay using the Bland-Altman plot at lower level. Nine healthy males were recruited and each provided four saliva samples on a single day to yield a total of 36 samples. Saliva samples were directly collected in plastic tubes using plastic straws, and subsequently pipetted onto cotton swabs (cotton saliva collection) and into clear sterile tubes (passive saliva collection). The melatonin levels were analyzed in duplicate using commercially available ELISA kits. The mean melatonin concentration in cotton saliva collection samples was significantly lower than that in passive saliva collection samples at higher melatonin level (>6 pg/mL). The Bland-Altman plot indicated that cotton swabs causes relative and proportional biases in the assay results. For lower melatonin level (<6 pg/mL), although the BA plots didn't show proportional and relative biases, there was no significant correlation between passive and cotton saliva collection samples. Our findings indicate an interference effect of cotton swabs on the assay result of salivary melatonin at lower melatonin level. Cotton-based collection devices might, thus, not be suitable for assessment of DLMO.

  20. Effects of saliva collection using cotton swabs on melatonin enzyme immunoassay

    Directory of Open Access Journals (Sweden)

    Katsuura Tetsuo

    2011-01-01

    Full Text Available Abstract Background Although various acceptable and easy-to-use devices have been used for saliva collection, cotton swabs are among the most common ones. Previous studies reported that cotton swabs yield a lower level of melatonin detection. However, this statistical method is not adequate for detecting an agreement between cotton saliva collection and passive saliva collection, and a test for bias is needed. Furthermore, the effects of cotton swabs have not been examined at lower melatonin level, a level at which melatonin is used for assessment of circadian rhythms, namely dim light melatonin onset (DLMO. In the present study, we estimated the effect of cotton swabs on the results of salivary melatonin assay using the Bland-Altman plot at lower level. Methods Nine healthy males were recruited and each provided four saliva samples on a single day to yield a total of 36 samples. Saliva samples were directly collected in plastic tubes using plastic straws, and subsequently pipetted onto cotton swabs (cotton saliva collection and into clear sterile tubes (passive saliva collection. The melatonin levels were analyzed in duplicate using commercially available ELISA kits. Results The mean melatonin concentration in cotton saliva collection samples was significantly lower than that in passive saliva collection samples at higher melatonin level (>6 pg/mL. The Bland-Altman plot indicated that cotton swabs causes relative and proportional biases in the assay results. For lower melatonin level ( Conclusion Our findings indicate an interference effect of cotton swabs on the assay result of salivary melatonin at lower melatonin level. Cotton-based collection devices might, thus, not be suitable for assessment of DLMO.

  1. Microcapillary blood sampling for serological examinations by radioimmunoassay (RIA) and enzyme immunoassay (ELISA)

    Energy Technology Data Exchange (ETDEWEB)

    Rodak, L.; Smid, B.; Valicek, L.; Jurak, E. (Vyzkumny Ustav Veterinarniho Lekarstvi, Brno-Medlanky (Czechoslovakia))

    1984-01-01

    Methods were tested of sampling blood and blood serum for serological examinations on filtration paper and into heparinized glass capillaries with transfer into the dilution solution of the given composition. Samples were also examined for ACH virus antibodies. The suitability of the sampling was verified by an examination of samples using ELISA and RIA methods. The results showed the suitability of sampling using microcapillaries. The titres of virus antibodies found using the ELISA and RIA methods were identical and the sensitivity of antibody detection was not reduced even after the sample had been stored for 60 days at a temperature of 20 degC.

  2. Microcapillary blood sampling for serological examinations by radioimmunoassay (RIA) and enzyme immunoassay (ELISA)

    International Nuclear Information System (INIS)

    Rodak, L.; Smid, B.; Valicek, L.; Jurak, E.

    1984-01-01

    Methods were tested of sampling blood and blood serum for serological examinations on filtration paper and into heparinized glass capillaries with transfer into the dilution solution of the given composition. Samples were also examined for ACH virus antibodies. The suitability of the sampling was verified by an examination of samples usiOg ELISA and RIA methods. The results showed the suitability of sampling using microcapillaries. The titres of virus antibodies found using the ELISA and RIA methods were identical and the sensitivity of antibody detection was not reduced even after the sample had been stored for 60 days at a temperature of 20 degC. (B.S.)

  3. Evaluation of Leishmania antigens preparation and storage for use in enzyme immunoassays

    Directory of Open Access Journals (Sweden)

    Celeste Beatriz Julieta

    1997-01-01

    Full Text Available The influence of time and temperature on the storage of an alkaline antigen of L.major-like and L.(V. braziliensis promastigotes added or not of a proteases inhibitor (PMSF was evaluted by means of an IgG-ELISA. Antibodies in assays using L. major-like antigen stored at -20oC for 6 monsths had a statistically lower geometric mean titer (GMT and different 95% confidence interval limits (CL than antigens stored otherwise, as assessed by the "t" statistic. The PMSF L. major-like antigen after storage for 6 months at a temperature of 4oC had the same GMT and 95% CL displayed at time zero as well as when storage for 4 and 6 months at -20oC. Significant diferences were not found when L.(V. braziliensis antigens were stored at times and temperatures mentioned; the PMSF antigen stored for 2 months at -70oC resulted in a lower serum GMT and 95% CL than any other, as assessed by the "t" statistic. Antigen performance did not show any statistical difference associated to the addition of PMSF within the same species; the largest difference between antigens was that between PMSF-L. (V. braziliensis and L. major-like without PMSF.

  4. The basis for decisions in the nuclear waste issue. Experiences of the legislative basis and the EIA process

    International Nuclear Information System (INIS)

    Keskitalo, Carina; Nordlund, Annika; Lindgren, Urban

    2009-03-01

    The purpose of the present study is to analyze the multi-level governance process in conjunction with the siting and design of a final repository for spent nuclear fuel. Since no similar activities have ever been implemented in Sweden, there is no available practice for how different laws should be coordinated and interpreted. The study sheds light on three general questions: 1) What is the formal decision-making mandate and what are the decision-making bodies at different levels (municipal level, regional or county level, and national level) according to the legislation, and what interpretation problems have these actors experienced with regard to the legislation and the EIA process? 2) What 'broader public' and organizations besides groups within the formal decision-making mandate have participated in the consultations, and what viewpoints have they expressed regarding the EIA process and consultations? 3) How have judgements and understanding of, and reactions to, risk related to the final repository been handled in the process? The study is mainly based on two different sources of material. A literature review with a focus on nuclear fuel management has been carried out within the social sciences field. Special interest has been devoted to discussing the content of legislation in relation to the EIA process and licensing, and the background and design of the EIA process. The EIA process is of special interest here, since it brings in both formal decision-making bodies and participation by broader groups. Furthermore, the literature review deals with theoretical perspectives regarding perceptions and communication of risk assessments. Literature reviews have also been conducted on minutes from EIA consultations during the period 2001 to 2007. The main source of the material used in the study is interview studies. The interview subjects represent both government authorities and non governmental organizations at the national, regional and local level. The results

  5. Development of a Nanobody-Based Lateral Flow Immunoassay for Detection of Human Norovirus.

    Science.gov (United States)

    Doerflinger, Sylvie Y; Tabatabai, Julia; Schnitzler, Paul; Farah, Carlo; Rameil, Steffen; Sander, Peter; Koromyslova, Anna; Hansman, Grant S

    2016-01-01

    Human noroviruses are the dominant cause of outbreaks of acute gastroenteritis. These viruses are usually detected by molecular methods, including reverse transcriptase PCR (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). Human noroviruses are genetically and antigenically diverse, with two main genogroups that are further subdivided into over 40 different genotypes. During the past decade, genogroup 2 genotype 4 (GII.4) has dominated in most countries, but recently, viruses belonging to GII.17 have increased in prevalence in a number of countries. A number of commercially available ELISAs and lateral flow immunoassays were found to have lower sensitivities to the GII.17 viruses, indicating that the antibodies used in these methods may not have a high level of cross-reactivity. In this study, we developed a rapid Nanobody-based lateral flow immunoassay (Nano-immunochromatography [Nano-IC]) for the detection of human norovirus in clinical specimens. The Nano-IC assay detected virions from two GII.4 norovirus clusters, which included the current dominant strain and a novel variant strain. The Nano-IC method had a sensitivity of 80% and specificity of 86% for outbreak specimens. Norovirus virus-like particles (VLPs) representing four genotypes (GII.4, GII.10, GII.12, and GII.17) could be detected by this method, demonstrating the potential in clinical screening. However, further modifications to the Nano-IC method are needed in order to improve this sensitivity, which may be achieved by the addition of other broadly reactive Nanobodies to the system. IMPORTANCE We previously identified a Nanobody (termed Nano-85) that bound to a highly conserved region on the norovirus capsid. In this study, the Nanobody was biotinylated and gold conjugated for a lateral flow immunoassay (termed Nano-IC). We showed that the Nano-IC assay was capable of detecting at least four antigenically distinct GII genotypes, including the newly emerging GII.17. In the clinical setting, the

  6. A New Microsphere-Based Immunoassay for Measuring the Activity of Transcription Factors

    Directory of Open Access Journals (Sweden)

    Tsai Chueh-Jen

    2010-01-01

    Full Text Available Abstract There are several traditional and well-developed methods for analyzing the activity of transcription factors, such as EMSA, enzyme-linked immunosorbent assay, and reporter gene activity assays. All of these methods have their own distinct disadvantages, but none can analyze the changes in transcription factors in the few cells that are cultured in the wells of 96-well titer plates. Thus, a new microsphere-based immunoassay to measure the activity of transcription factors (MIA-TF was developed. In MIA-TF, NeutrAvidin-labeled microspheres were used as the solid phase to capture biotin-labeled double-strand DNA fragments which contain certain transcription factor binding elements. The activity of transcription factors was detected by immunoassay using a transcription factor-specific antibody to monitor the binding with the DNA probe. Next, analysis was performed by flow cytometry. The targets hypoxia-inducible factor-1α (HIF-1α and nuclear factor-kappa B (NF-κB were applied and detected in this MIA-TF method; the results that we obtained demonstrated that this method could be used to monitor the changes of NF-κB or HIF within 50 or 100 ng of nuclear extract. Furthermore, MIA-TF could detect the changes in NF-κB or HIF in cells that were cultured in wells of a 96-well plate without purification of the nuclear protein, an important consideration for applying this method to high-throughput assays in the future. The development of MIA-TF would support further progress in clinical analysis and drug screening systems. Overall, MIA-TF is a method with high potential to detect the activity of transcription factors.

  7. Development of a lateral flow immunoassay for rapid diagnosis of potato blackleg caused by Dickeya species.

    Science.gov (United States)

    Safenkova, Irina V; Zaitsev, Ilya A; Varitsev, Yuri A; Byzova, Nadezhda A; Drenova, Natalia V; Zherdev, Anatoly V; Dzantiev, Boris B

    2017-03-01

    Early detection of potato infections is essential for effective disease management. The aim of this study was to develop a lateral flow immunoassay (LFIA) for rapid detection of a serious potato disease, potato blackleg, caused by Dickeya dianthicola and Dickeya solani. Polyclonal antibodies specific to different strains of Dickeya were obtained from rabbits after immunization with bacterial cells of D. dianthicola and D. solani. Enzyme-linked immunosorbent assay testing with use of a wide range of bacterial species showed that the polyclonal antibodies detect closely related strains of D. dianthicola and D. solani. Cross-reactivity with widespread pathogenic bacteria (nine species) and saprophytes of healthy potato plants was not detected. The LFIA based on the obtained antibodies and gold nanoparticles with average diameter of 20 nm was developed. Under optimized conditions, the LFIA method enabled the analysis of potato extracts within 10 min, with a visual limit of detection of 1 × 10 5  CFU/ml for leaves and 4 × 10 5  CFU/ml for tubers. The assay was tested on potato stem and tuber extracts, and the results of the LFIA were confirmed in 92.1% of samples using the real-time polymerase chain reaction. The findings confirmed that the developed LFIA could be used for monitoring blackleg infection without the need for special equipment or skills. Graphical Abstract The developed lateral flow immunoassay is an efficient tool for rapid detection of a serious potato disease, potato blackleg, caused by Dickeya dianthicola and Dickeya solani.

  8. Nanogold-polyaniline-nanogold microspheres-functionalized molecular tags for sensitive electrochemical immunoassay of thyroid-stimulating hormone

    Energy Technology Data Exchange (ETDEWEB)

    Cui Yuling; Chen Huafeng; Hou Li; Zhang Bing; Liu Bingqian; Chen Guonan [Ministry of Education Key Laboratory of Analysis and Detection for Food Safety, Fujian Province Key Laboratory of Analysis and Detection for Food Safety, Department of Chemistry and Chemical Engineering, Fuzhou University, Fuzhou 350108 (China); Tang Dianping, E-mail: dianping.tang@fzu.edu.cn [Ministry of Education Key Laboratory of Analysis and Detection for Food Safety, Fujian Province Key Laboratory of Analysis and Detection for Food Safety, Department of Chemistry and Chemical Engineering, Fuzhou University, Fuzhou 350108 (China)

    2012-08-13

    Highlights: Black-Right-Pointing-Pointer A novel immunosensing strategy was designed for detection of thyroid-stimulating hormone. Black-Right-Pointing-Pointer Using nanogold-polyaniline-nanogold microspheres as molecular tags. Black-Right-Pointing-Pointer Improvement of electrochemical activity of nanolabels. Black-Right-Pointing-Pointer Combination enzyme labels with nanolabels for signal amplification. - Abstract: Methods based on nanomaterial labels have been developed for electrochemical immunosensors and immunoassays, but most involved low sensitivity. Herein a novel class of molecular tags, nanogold-polyaniline-nanogold microspheres (GPGs), was first synthesized and functionalized with horseradish peroxidase-conjugated thyroid-stimulating hormone antibody (HRP-Ab{sub 2}) for sensitive electrochemical immunoassay of thyroid-stimulating hormone (TSH). X-ray diffraction, confocal Raman spectroscopy, scanning electron microscope and transmission electron microscope were employed to characterize the prepared GPGs. Based on a sandwich-type immunoassay format, the assay was performed in pH 5.0 acetate buffer containing 6.0 mmol L{sup -1} H{sub 2}O{sub 2} by using GPG-labeled HRP-Ab{sub 2} as molecular tags. Compared with pure polyaniline nanospheres and gold nanoparticles alone, the GPG hybrid nanostructures increased the surface area of the nanomaterials, and enhanced the immobilized amount of HRP-Ab{sub 2}. Several labeling protocols comprising HRP-Ab{sub 2}, nanogold particle-labeled HRP-Ab{sub 2}, and polyaniline nanospheres-labeled HRP-Ab{sub 2}, were also investigated for determination of TSH and improved analytical features were obtained by using the GPG-labeled HRP-Ab{sub 2}. With the GPG labeling method, the effects of incubation time and pH of acetate buffer on the current responses of the immunosensors were also studied. The strong attachment of HRP-Ab{sub 2} to the GPGs resulted in a good repeatability and intermediate precision down to 7%. The dynamic

  9. Rapid, automated, parallel quantitative immunoassays using highly integrated microfluidics and AlphaLISA

    Science.gov (United States)

    TakYu, Zeta; Guan, Huijiao; Ki Cheung, Mei; McHugh, Walker M.; Cornell, Timothy T.; Shanley, Thomas P.; Kurabayashi, Katsuo; Fu, Jianping

    2015-06-01

    Immunoassays represent one of the most popular analytical methods for detection and quantification of biomolecules. However, conventional immunoassays such as ELISA and flow cytometry, even though providing high sensitivity and specificity and multiplexing capability, can be labor-intensive and prone to human error, making them unsuitable for standardized clinical diagnoses. Using a commercialized no-wash, homogeneous immunoassay technology (‘AlphaLISA’) in conjunction with integrated microfluidics, herein we developed a microfluidic immunoassay chip capable of rapid, automated, parallel immunoassays of microliter quantities of samples. Operation of the microfluidic immunoassay chip entailed rapid mixing and conjugation of AlphaLISA components with target analytes before quantitative imaging for analyte detections in up to eight samples simultaneously. Aspects such as fluid handling and operation, surface passivation, imaging uniformity, and detection sensitivity of the microfluidic immunoassay chip using AlphaLISA were investigated. The microfluidic immunoassay chip could detect one target analyte simultaneously for up to eight samples in 45 min with a limit of detection down to 10 pg mL-1. The microfluidic immunoassay chip was further utilized for functional immunophenotyping to examine cytokine secretion from human immune cells stimulated ex vivo. Together, the microfluidic immunoassay chip provides a promising high-throughput, high-content platform for rapid, automated, parallel quantitative immunosensing applications.

  10. Evaluation of four immunoassays for diagnosis of brucellosis in Cuba

    International Nuclear Information System (INIS)

    Peraza, C.; Valdes, O.; Fonseca, N.; Garcia, M.; Alvarez, M.; Izquierdo, D.L.

    1998-01-01

    Four immunoassays (two indirect and two competitive ones) were evaluated by samples from areas free of disease, free by vaccination and affected areas using as reference techniques the Bengal Rose Tests, the Antigen in Buffered Plate Tests and the Complement Fixation Reaction Test. The evaluated samples demonstrated that the competitive assays (ELISAC-1 and ELISAC-2) detected less false positives than the indirect ones (ELISAI-1 and ELISAI-2). Of the competitive ELISAS, version 2 presented better sensitivity and specificity results in affected areas for 95% confidence: 80.9 - 96.9% and 97.5 - 99.4% respectively with positive predictive value in the range of 76 to 94% and negative predictive one between 98.1 and 99.7%. It was concluded that this assay can be used for brucellosis control because it gives higher assurance than the other evaluated immunoassays and it can discriminate infected from vaccinated animals. (author)

  11. Indigenous Peoples Involment At The Environmental Impact Assessment (EIA) Process in Tabi Mamta Area Of Papua Province

    Science.gov (United States)

    Dhiksawan, Ferdinand Saras; Hadi, Sudharto P.; Samekto, Adji; Sasongko, Dwi P.

    2018-02-01

    The purpose of this study is to find a picture of the involvement of Indigenous Peoples of Tabi Mamta in the process of environmental impact assessment (EIA) in Tabi Mamta customary territory. The method and type of research used is non-ethnographic qualitative research with data collection techniques using limited observation techniques. Data and information in the field will be analyzed using constructivism paradigm. The paradigm of constructivism is based on an interpretive understanding called hermeneutics (hermeneuien) in the sense of interpreting, giving understanding, translating data and information obtained in the research location as a result of social reality. The results of this study indicate that the customary community of Tabi Mamta is a unit of customary community that still has territorial customary territory, has a customary leadership structure, still visible relationship of kinship, cultural values as well as customary norms and sanctions, and has environmental wisdom in maintaining existence Natural resources. In the socio-cultural system of customary communities there are components such as customary stratification, permissiveness, communication, reciprocity, past history, cultural values, customary norms and sanctions, religious and customary leadership. Components in the socio-cultural system of indigenous and tribal peoples play a role in the EIA process in the Tabi Mamta customary area especially in the environmental feasibility decision making process. The components of custom stratification, cultural values and customary norms play a role in the EIA process. In customary stratification there is uncustomary structure in the ondoafi, Iram and Tribal Leadership. Components in a sociual cultural system is a unity resulting from interaction between individuals and groups to prevent environmental damage and disturbance of natural resources. Natural resources are considered as ancestral symbols passed down by ancestors from generation to

  12. Indigenous Peoples Involment At The Environmental Impact Assessment (EIA Process in Tabi Mamta Area Of Papua Province

    Directory of Open Access Journals (Sweden)

    Saras Dhiksawan Ferdinand

    2018-01-01

    Full Text Available The purpose of this study is to find a picture of the involvement of Indigenous Peoples of Tabi Mamta in the process of environmental impact assessment (EIA in Tabi Mamta customary territory. The method and type of research used is non-ethnographic qualitative research with data collection techniques using limited observation techniques. Data and information in the field will be analyzed using constructivism paradigm. The paradigm of constructivism is based on an interpretive understanding called hermeneutics (hermeneuien in the sense of interpreting, giving understanding, translating data and information obtained in the research location as a result of social reality. The results of this study indicate that the customary community of Tabi Mamta is a unit of customary community that still has territorial customary territory, has a customary leadership structure, still visible relationship of kinship, cultural values as well as customary norms and sanctions, and has environmental wisdom in maintaining existence Natural resources. In the socio-cultural system of customary communities there are components such as customary stratification, permissiveness, communication, reciprocity, past history, cultural values, customary norms and sanctions, religious and customary leadership. Components in the socio-cultural system of indigenous and tribal peoples play a role in the EIA process in the Tabi Mamta customary area especially in the environmental feasibility decision making process. The components of custom stratification, cultural values and customary norms play a role in the EIA process. In customary stratification there is uncustomary structure in the ondoafi, Iram and Tribal Leadership. Components in a sociual cultural system is a unity resulting from interaction between individuals and groups to prevent environmental damage and disturbance of natural resources. Natural resources are considered as ancestral symbols passed down by ancestors from

  13. Nanobody Based Immunoassay for Human Soluble Epoxide Hydrolase Detection Using Polymeric Horseradish Peroxidase (PolyHRP) for Signal Enhancement: The Rediscovery of PolyHRP?

    Science.gov (United States)

    Li, Dongyang; Cui, Yongliang; Morisseau, Christophe; Gee, Shirley J; Bever, Candace S; Liu, Xiangjiang; Wu, Jian; Hammock, Bruce D; Ying, Yibin

    2017-06-06

    Soluble epoxide hydrolase (sEH) is a potential pharmacological target for treating hypertension, vascular inflammation, cancer, pain, and multiple cardiovascular related diseases. A variable domain of the heavy chain antibody (termed single domain antibody (sdAb), nanobody, or VHH) possesses the advantages of small size, high stability, ease of genetic manipulation, and ability for continuous manufacture, making such nanobody a superior choice as an immunoreagent. In this work, we developed an ultrasensitive nanobody based immunoassay for human sEH detection using polymeric horseradish peroxidase (PolyHRP) for signal enhancement. Llama nanobodies against human sEH were used as the detection antibody in sandwich enzyme linked immunosorbent assays (ELISA) with polyclonal anti-sEH as the capture antibody. A conventional sandwich ELISA using a horseradish peroxidase (HRP) labeled anti-hemeagglutinin (HA) tag as the tracer showed a marginal sensitivity (0.0015 optical density (OD)·mL/ng) and limit of detection (LOD) of 3.02 ng/mL. However, the introduction of the PolyHRP as the tracer demonstrated a 141-fold increase in the sensitivity (0.21 OD·mL/ng) and 57-fold decrease in LOD (0.05 ng/mL). Systematic comparison of three different tracers in four ELISA formats demonstrated the overwhelming advantage of PolyHRP as a label for nanobody based immunoassay. This enhanced sEH immunoassay was further evaluated in terms of selectivity against other epoxide hydrolases and detection of the target protein in human tissue homogenate samples. Comparison with an enzyme activity based assay and a Western blot for sEH detection reveals good correlation with the immunoassay. This work demonstrates increased competiveness of nanobodies for practical sEH protein detection utilizing PolyHRP. It is worthwhile to rediscover the promising potential of PolyHRP in nanobody and other affinity based methods after its low-profile existence for decades.

  14. A specific radio-immunoassay for Gonadotrophin-releasing hormone

    International Nuclear Information System (INIS)

    Hendricks, S.; Millar, R.; Pimstone, B.

    1975-01-01

    A specific antiserum has been made to synthetic gonadotrophin-releasing hormone (GnRH) conjugated to keyhole limpet haemocyanin and appears to be directed against amino acids 6 - 8 of this decapeptide. This has allowed the development of a radio-immunoassay for GnRH sensitive to 5 picograms per tube. Although it is easily measurable in hypothalamic extracts, we have failed to detect GnRH in plasma and urine from normal subjects and menopausal women

  15. Cavity enhanced immunoassay measurements in microtiter plates using BBCEAS

    OpenAIRE

    Bajuszova, Z; Ali, Z; Scott, SM; Seetohul, LN; Islam, M

    2016-01-01

    We report on the first detailed use of broadband cavity enhanced absorption spectroscopy (BBCEAS) as a detection system for immunoassay. A vertical R ≥ 0.99 optical cavity was integrated with a motorised XY stage, which functioned as a receptacle for 96 well microtiter plates. The custom built cavity enhanced microplate reader was used to make measurements on a commercially available osteocalcin sandwich ELISA kit. A 30 fold increase in path length was obtained with a minimum detectable chang...

  16. Efavirenz interference in urine screening immunoassays for tetrahydrocannabinol.

    Science.gov (United States)

    Oosthuizen, Nicholette M; Laurens, Johannes B

    2012-03-01

    It has been known for some time that the antiretroviral drug, efavirenz (EFV), cross-reacts in urine immunoassays for tetrahydrocannabinol (THC). Because published studies investigating this phenomenon are limited, cross-reactivity information for several immunoassays is lacking. Reports of possible false-positive THC results from clinicians conducting workplace testing prompted us to investigate cross-reactivity for assays frequently employed in our own setting. In light of the potentially deleterious consequences of misclassification, information about EFV cross-reactivity should be included in product information to facilitate interpretation of results and assay selection. Random urine samples from 30 patients on EFV therapy were analysed for THC metabolites by two near-testing devices (THC One Step Marijuana and Rapid Response(®) Drugs of Abuse Test Strips) and two automated immunoassays (Roche Diagnostics Cannabinoids II and Beckman Coulter SYNCHRON(®) Systems THC2). THC confirmatory testing was performed by gas chromatography-mass spectrometry (GC-MS). GC-MS failed to detect THC metabolites in any of the samples, as did three of the four immunoassays. However, the Rapid Response(®) test strips yielded positive results in 28 out of 30 samples, which could be reversed on re-testing after sample pretreatment with glucuronidase. Our study supports previous findings that interference is attributable to a glucuronidated EFV metabolite. We postulate that cross-reactivity is influenced by the composition of immunogens used to elicit anti-THC antibodies. Since access to such information is restricted, contributions from scientists in the antibody industry may be enlightening.

  17. History of inductively coupled plasma mass spectrometry-based immunoassays

    International Nuclear Information System (INIS)

    Giesen, Charlotte; Waentig, Larissa; Panne, Ulrich; Jakubowski, Norbert

    2012-01-01

    The analysis of biomolecules requires highly sensitive and selective detection methods capable of tolerating a complex, biological matrix. First applications of biomolecule detection by ICP-MS relied on the use of heteroelements as a label for quantification. However, the combination of immunoassays and ICP-MS facilitates multiparametric analyses through elemental tagging, and provides a powerful alternative to common bioanalytical methods. This approach extends the detection of biomarkers in clinical diagnosis, and has the potential to provide a deeper understanding of the investigated biological system. The results might lead to the detection of diseases at an early stage, or guide treatment plans. Immunoassays are well accepted and established for diagnostic purposes, albeit ICP-MS is scarcely applied for the detection of immune-based assays. However, the screening of biomarkers demands high throughput and multiplex/multiparametric techniques, considering the variety of analytes to be queried. Finally, quantitative information on the expression level of biomarkers is highly desirable to identify abnormalities in a given organism. Thus, it is the aim of this review to introduce the fundamentals, and to discuss the enormous strength of ICP-MS for the detection of different immunoassays on the basis of selected applications, with a special focus on LA‐ICP‐MS. - Highlights: ► We discuss the fundamentals of elemental tagging for ICP‐MS applications. ► We propose a definition for the expressions “label” and “tag”. ► We highlight LA‐ICP‐MS‐based heteroelement detection. ► We give an historic overview on ICP-MS and LA‐ICP‐MS-based immunoassays. ► In a personal outlook, we discuss future improvements realistically attainable.

  18. Label-Free Electrochemical Immunoassay for C-Reactive Protein

    Directory of Open Access Journals (Sweden)

    Madasamy Thangamuthu

    2018-03-01

    Full Text Available C-reactive protein (CRP is one of the most expressed proteins in blood during acute phase inflammation, and its minute level increase has also been recognized for the clinical diagnosis of cardio vascular diseases. Unfortunately, the available commercial immunoassays are labour intensive, require large sample volumes, and have practical limitations, such as low stability and high production costs. Hence, we have developed a simple, cost effective, and label-free electrochemical immunoassay for the measurement of CRP in a drop of serum sample using an immunosensor strip made up of a screen printed carbon electrode (SPE modified with anti-CRP functionalized gold nanoparticles (AuNPs. The measurement relies on the decrease of the oxidation current of the redox indicator Fe3+/Fe2+, resulting from the immunoreaction between CRP and anti-CRP. Under optimal conditions, the present immunoassay measures CRP in a linear range from 0.4–200 nM (0.047–23.6 µg mL−1, with a detection limit of 0.15 nM (17 ng mL−1, S/N = 3 and sensitivity of 90.7 nA nM−1, in addition to a good reproducibility and storage stability. The analytical applicability of the presented immunoassay is verified by CRP measurements in human blood serum samples. This work provides the basis for a low-priced, safe, and easy-to-use point-of-care immunosensor assay to measure CRP at clinically relevant concentrations.

  19. History of inductively coupled plasma mass spectrometry-based immunoassays

    Energy Technology Data Exchange (ETDEWEB)

    Giesen, Charlotte, E-mail: charlotte.giesen@uzh.ch [University of Zurich, Institute of Molecular Life Sciences, Winterthurerstrasse 190, 8057 Zurich (Switzerland); Waentig, Larissa [BAM Federal Institute for Materials Research and Testing, Richard-Willstaetter-Strasse 11, 12489 Berlin (Germany); Panne, Ulrich [BAM Federal Institute for Materials Research and Testing, Richard-Willstaetter-Strasse 11, 12489 Berlin (Germany); Humboldt-Universitaet zu Berlin, Department of Chemistry, Brook-Taylor-Strasse 2, 12489 Berlin (Germany); Jakubowski, Norbert [BAM Federal Institute for Materials Research and Testing, Richard-Willstaetter-Strasse 11, 12489 Berlin (Germany)

    2012-10-15

    The analysis of biomolecules requires highly sensitive and selective detection methods capable of tolerating a complex, biological matrix. First applications of biomolecule detection by ICP-MS relied on the use of heteroelements as a label for quantification. However, the combination of immunoassays and ICP-MS facilitates multiparametric analyses through elemental tagging, and provides a powerful alternative to common bioanalytical methods. This approach extends the detection of biomarkers in clinical diagnosis, and has the potential to provide a deeper understanding of the investigated biological system. The results might lead to the detection of diseases at an early stage, or guide treatment plans. Immunoassays are well accepted and established for diagnostic purposes, albeit ICP-MS is scarcely applied for the detection of immune-based assays. However, the screening of biomarkers demands high throughput and multiplex/multiparametric techniques, considering the variety of analytes to be queried. Finally, quantitative information on the expression level of biomarkers is highly desirable to identify abnormalities in a given organism. Thus, it is the aim of this review to introduce the fundamentals, and to discuss the enormous strength of ICP-MS for the detection of different immunoassays on the basis of selected applications, with a special focus on LA-ICP-MS. - Highlights: Black-Right-Pointing-Pointer We discuss the fundamentals of elemental tagging for ICP-MS applications. Black-Right-Pointing-Pointer We propose a definition for the expressions 'label' and 'tag'. Black-Right-Pointing-Pointer We highlight LA-ICP-MS-based heteroelement detection. Black-Right-Pointing-Pointer We give an historic overview on ICP-MS and LA-ICP-MS-based immunoassays. Black-Right-Pointing-Pointer In a personal outlook, we discuss future improvements realistically attainable.

  20. Environmental impact assessment modern dressed? To the amendment of the EIA act and other acts and regulations

    International Nuclear Information System (INIS)

    Feldmann, Ulrike

    2017-01-01

    On 22 December 2016, the Federal Ministry for the Environment (BMUB) presented the ''Draft Act for the Modernization of the Act on the Environmental Impact Assessment'' within the framework of the association consultation, as well as the ''Draft first Ordinance Amending the Ordinance on the Approval Procedure - 9. BImSchV''. The EIA Modernization Act as well as the Atomic Act Procedure Regulation and the Federal Mining Act should be revised by terms of an omnibus act. The association consultation was held on 18 January 2017.

  1. Current status and future developments in radiolabelled immunoassays

    International Nuclear Information System (INIS)

    Edwards, R.

    1998-01-01

    Radioisotopes are used extensively in medical practice and their use in RIA or IRMA usually represent a small proportion of the total. Radiolabelled immunoassays based on 125 I constitute a simple didactic, cost effective and robust technology which is still regarded as the reference method in many clinical applications. The IAEA has implemented many successful programmes using the ''bulk reagent'' approach, involving 68 countries in all the different regions. The main achievements have been in technology transfer with self sufficiency in production for some countries; training of large numbers of staff; quality control and quality assurance schemes; devolution of screening programmes for neonatal congenital hypothryoidism. Alternatives to the use of radioisotopic tracers are constrained by many factors and are often only available in restricted commercial packages. They are often not suitable for technology transfer programmes and often lack any didactic component in addition to a relative high cost. The production of radiolabels using 125 I is both simple and adaptable. In addition expertise in their preparation and purification is widespread even in developing countries. Together with the ease of producing antibodies, the facts have made 125 I-radiolabelled immunoassays ideal for investigative procedures for many research activities (30,31) particularly in the medical context where radioisotopes are commonly used. In conclusion, even a superficial examination of public health statistics for various countries throughout the continents indicates a need for a simple, inexpensive and robust analytical tool. In this light, there is a predicted continuing role for radiolabelled immunoassays. (author)

  2. Simple patterned nanofiber scaffolds and its enhanced performance in immunoassay.

    Directory of Open Access Journals (Sweden)

    Jing Wang

    Full Text Available Cancer has become the leading cause of death worldwide; early diagnosis and treatment of cancers is critical for the survival of the patients. The concentration of cancer markers in easy-to-access biological fluids can provide great assistance in screening for occult primary cancers, distinguishing malignant from benign findings, determining prognosis and prediction for cancer patients. The multiplex detection technology of a panel of cancer markers can greatly increase the accuracy of disease diagnosis. Herein, we briefly fabricate a high-throughput micro-immunoassay based on the electrospun polystyrene (PS substrates to improve detection sensitivity. The immunoassay was evaluated by analyzing three different cancer biomarkers (AFP, CEA, VEGF. For AFP, CEA, VEGF immunofluorescence assay, the LOD of assay conducted on electrospun PS substrates before or after plasma and the conventional PS substrates were 0.42, 0.10, 1.12 ng/mL, 0.57, 0.09, 1.24 ng/mL, and 159.75, 26.19, 385.59 pg/mL, respectively (P < 0.05. Due to the high porosity and large surface area-to-volume ratio which is the foremost merit of nanostructures, and the plasma treatment which make the hydrophobic PS nanofibers hydropholic, the nanofibers substrates showed sufficient retention of immunoassay functionality and high potential for capture molecules immobilization. Consequently, the immunofluorescence assay conducted on electrospun PS substrates could significantly enhance the sensitivity and limits of detection.

  3. Ultrasensitive immunoassay based on electrochemical measurement of enzymatically produced polyaniline.

    Science.gov (United States)

    Lai, Guosong; Zhang, Haili; Tamanna, Tasnuva; Yu, Aimin

    2014-02-04

    A novel ultrasensitive immunoassay method was developed based on the electrochemical measurement of polyaniline, which was catalytically produced by horseradish peroxidase-functionalized gold nanoparticle (HRP-Au NP) probe at an immunosensor. The immunosensor was prepared step-wise by first modifying the electrode with reduced graphene oxide (rGO)/Au NPs nanocomposite followed by the immobilization of capture antibodies on its surface. After performing a sandwich immunoreaction, the quantitatively captured HRP-Au NP nanoprobes could catalyze oxidation of aniline to produce electroactive polyaniline on the immunosensor surface. The electrochemical measurement of polyaniline enabled a novel detection strategy for HRP-based immunoassay. Both the signal amplification of the HRP-Au NP nanoprobe and the electron transfer acceleration of rGO/Au NPs on the immunosensor surface greatly improved the detection sensitivity of the immunoassay method. With the use of human IgG as a model analyte, this method showed a wide linear range over 4 orders of magnitude with a detection limit of 9.7 pg/mL. In addition, the immunosensor had low cost, satisfactory reproducibility and stability, and acceptable reliability. The relatively positive potential range for the polyaniline measurement completely excluded the conventional interference from dissolved oxygen. Thus, this method provides a promising potential for practical applications.

  4. A análise dos impactos socioeconômicos na estruturação do EIA/RIMA: a importância da Abordagem Humanista para a Sustentabilidade The analysis of socio-economic impacts on the structure of EIA / RIMA: the importance of humanist approach to sustainability

    Directory of Open Access Journals (Sweden)

    Leandro Carlos Dias Conde

    2013-03-01

    Full Text Available DOI: 10.5902/198346597686O presente trabalho trata da defesa da abordagem humanista na estruturação da análise de Impacto Ambiental (EIA e do seu Relatório de Impacto Ambiental (RIMA, no que concerne aos impactos socioeconômicos abordados, por compreender que a abordagem convencional não dá conta das demandas sociais, pois somente esta nova abordagem dará, de fato, o caráter sustentável da EIA/RIMA. The present work deals with the defense of the humanistic approach in structuring the analysis of Environmental Impact Assessment (EIA and its Environmental Impact Report (RIMA in relation to socioeconomic impacts addressed to realize that the conventional approach does not account social demands, as only this new approach will in fact the sustainable nature of the EIA / RIMA.

  5. Functionalization of single-walled carbon nanotubes with protein by click chemistry as sensing platform for sensitized electrochemical immunoassay

    International Nuclear Information System (INIS)

    Qi Honglan; Ling Chen; Huang Ru; Qiu Xiaoying; Shangguan Li; Gao Qiang; Zhang Chengxiao

    2012-01-01

    Highlights: ► Single-walled carbon nanotubes were functionalized with protein by click chemistry. ► The SWNTs conjugated with protein showed excellent dispersion in water and kept good bioacitvity. ► A competitive electrochemical immunoassay for the determination of anti-IgG was developed with high sensitivity and good stability. - Abstract: The application of the Cu(I)-catalyzed [3 + 2] Huisgen cycloaddition to the functionalization of single-walled carbon nanotubes (SWNTs) with the protein and the use of the artificial SWNTs as a sensing platform for sensitive immunoassay were reported. Covalent functionalization of azide decorated SWNTs with alkyne modified protein was firstly accomplished by the Cu(I)-catalyzed [3 + 2] Huisgen cycloaddition. FT-IR spectroscopy, Raman spectroscopy, X-ray photoelectron spectroscopy, scanning electron microscopy and transmission electron micrograph were used to characterize the protein-functionalized SWNTs. It was found that the SWNTs conjugated with the proteins showed excellent dispersion in water and kept good bioacitivity when immunoglobulin (IgG) and horseradish peroxidase (HRP) were chosen as model proteins. As a proof-of-concept, IgG-functionalized SWNTs were immobilized onto the surface of a glassy carbon electrode by simple casting method as immunosensing platform and a sensitive competitive electrochemical immunoassay was developed for the determination of anti-immunoglobulin (anti-IgG) using HRP as enzyme label. The fabrication of the immunosensor were characterized by cyclic voltammetry and electrochemical impedance spectroscopy with the redox probe [Fe(CN) 6 ] 3−/4− . The SWNTs as immobilization platform showed better sensitizing effect, a detection limit of 30 pg mL −1 (S/N = 3) was obtained for anti-IgG. The proposed strategy provided a stable immobilization method and sensitized recognition platform for analytes. This work demonstrated that the click coupling of SWNTs with protein was an effective

  6. Testing the usability of the Rapid Impact Assessment Matrix (RIAM) method for comparison of EIA and SEA results

    International Nuclear Information System (INIS)

    Kuitunen, Markku; Jalava, Kimmo; Hirvonen, Kimmo

    2008-01-01

    This study examines how the results of Environmental Impact Assessment (EIA) and Strategic Environmental Assessment (SEA) could be compared using the Rapid Impact Assessment Matrix (RIAM) method. There are many tools and techniques that have been developed for use in impact assessment processes, including scoping, checklists, matrices, qualitative and quantitative models, literature reviews, and decision-support systems. While impact assessment processes have become more technically complicated, it is recognized that approaches including simpler applications of available tools and techniques are also appropriate. The Rapid Impact Assessment Matrix (RIAM) is a tool for organizing, analysing and presenting the results of a holistic EIA. RIAM was originally developed to compare the impact of alternative procedures in a single project. In this study, we used RIAM to compare the environmental and social impact of different projects, plans and programs realized within the same geographical area. RIAM scoring is based on five separate criteria. The RIAM criteria were applied to the impact that was considered to be the most significant in the evaluated cases, and scores were given both on environmental and social impact. Our results revealed that the RIAM method could be used for comparison and ranking of separate and distinct projects, plans, programs and policies, based on their negative or positive impact. Our data included 142 cases from the area of Central Finland that is covered by the Regional Council of Central Finland. This sample consisted of various types of projects, ranging from road construction to education programs that applied for EU funding

  7. [Clinical value of fluorescence lateral flow immunoassay in diagnosis of influenza A in children].

    Science.gov (United States)

    Guo, Chun; Zhong, Li-Li; Yi, Hong-Ling; Chen, Min

    2016-12-01

    To evaluate the clinical value of a new type of fluorescence lateral flow immunoassay in rapid detection of influenza A virus. A total of 378 samples of nasopharyngeal secretions were collected from 378 children with influenza-like symptoms to detect the influenza A virus by fluorescence lateral flow immunoassay, colloidal gold immunoassay, and RT-PCR between July 2015 and August 2015. Of the 378 samples, 81 (21.4%) were positive for influenza A virus by RT-PCR. Compared with RT-PCR, the sensitivities of fluorescence lateral flow immunoassay and colloidal gold immunoassay were 90.1% (73/81) and 75.3% (61/81), respectively, and the specificities were 99.3% (295/297) and 98.3% (292/297), respectively. The average threshold cycle (Ct) value for the positive samples detected by the fluorescence lateral flow immunoassay (30.6) was higher than that for the positive samples detected by the colloidal gold immunoassay (28.7). Compared with colloidal gold immunoassay, fluorescence lateral flow immunoassay has higher sensitivity, specificity, and concordance rate with RT-PCR, suggesting that it can be used for early screening and diagnosis of influenza A.

  8. Production of polyclonal antibody against madecassoside and development of immunoassay methods for analysis of triterpene glycosides in Centella asiatica.

    Science.gov (United States)

    Tassanawat, Patcharin; Putalun, Waraporn; Yusakul, Gorawit; Sritularak, Boonchoo; Juengwatanatrakul, Thaweesak; Tanaka, Hiroyuki

    2013-01-01

    Centella asiatica (L.) Urban consists of two major triterpene glycosides, asiaticoside (AS) and madecassoside (MA), as active components used for wound healing and enhancing memory. To produce a polyclonal antibody against madecassoside (MA-PAb) and develop enzyme-linked immunosorbent assay (ELISA) and Eastern blotting methods for quantitative analysis of triterpene glycosides in Centella asiatica. An ELISA method was developed using polyclonal antibody against MA. An Eastern blotting method on the PES membrane was established for determination of MA and AS. The immunoassays were validated for sensitivity, precision, specificity and accuracy. The prepared MA-PAb shows specificity to MA and AS. The measuring range of triterpene glycosides was 0.39-50 µg/mL using the ELISA method. An Eastern blotting method was developed for determining individual MA and AS, which could be detected in the range of 62.5-500 ng. The limit of detection for MA and AS was 31.25 ng. The two methods developed showed good specificity, precision, and accuracy, and also correlated with high-performance liquid chromatography. These immunoassays have several advantages that include high sensitivity as well as being rapid and facile for determination of the triterpene glycosides in C. asiatica. Copyright © 2012 John Wiley & Sons, Ltd.

  9. Los Sistemas de Información Geográfica: una herramienta de análisis en los Estudios de Impacto Ambiental (EIA)

    OpenAIRE

    Rodríguez Jaume, María José

    2001-01-01

    En este capítulo se presenta una herramienta de trabajo que si bien hoy ya no puede ser tachada de novedosa sí que está viviendo en España su momento de máxima aceptación. Los Sistemas de Información Geográfica (SIGs) entroncan directamente con la filosofía y aspiraciones anheladas con la práctica del desarrollo sostenible o ecodesarrollo en general, y en particular, con los Estudios de Impacto Ambiental (EIA). Los SIGs palian algunas de las deficiencias y limitaciones de los EIA al uso. En c...

  10. Spatio-temporal characteristics of the Equatorial Ionization Anomaly (EIA) in the East African region via ionospheric tomography during the year 2012

    Science.gov (United States)

    Kassa, T.; Damtie, B.; Bires, A.; Yizengaw, E.; Cilliers, P.

    2015-01-01

    We present the characteristics of the EIA in the East African sector inferred from ground-based GPS receivers via ionospheric tomography during the year 2012. For the analysis, we developed and used a 2D ionospheric tomography imaging software based on Bayesian inversion approach. To reconstruct ionospheric electron density form slant Total Electron Content (sTEC) measurements, we selected a chain of ten ground-based GPS receivers with stations' codes and geomagnetic coordinates: ARMI (3.03 °S, 109.29 °E), DEBK (4.32 °N, 109.48 °E), ASOS (1.14 °N, 106.16 °E), NEGE (3.60 °S, 111.35 °E), SHIS (3.26 °N, 110.62 °E), ASAB (4.91 °N, 114.34 °E), SHEB (7.36 °N, 110.60 °E), EBBE (9.54 °S, 104.10 °E), DODM (16.03 °S, 109.04 °E) & NAMA (11.49 °N, 113.60 °E). The temporal, spatial and storm-time characteristics of the EIA and the hourly, day-to-day and seasonal variations of the maximum electron density of F2 region (NmF2) at 15.29°S geomagnetic latitude are presented. We found that the magnitude of the peak and the width/thickness of the EIA pronounced during the equinox and weakened during the solstice seasons at 2100 LT. It is also observed that the EIA persisted for longer time in equinox season than the solstice season. The spatial appearance of the northern and southern anomalies are observed starting from 6.12 ° N and 10 ° S respectively along geomagnetic latitude during equinox season. The EIA is localized between 180 km and 450 km along the altitude during December solstice. The analysis on the NmF2 demonstrated a significant dependence on local time, day and season of the year. We also investigated the storm response of the EIA for the magnetic storm of Day Of the Year (DOY) 274-276. It is observed that the disturbance dynamo related composition change (O/N2 ratio) resulted in a well-developed EIA with an increase in the peak and the width of the EIA at 2100 LT on DOY 275 (main phase of the storm) compared to 274 (initial phase of the storm

  11. A novel fluorescence immunoassay for the sensitive detection of Escherichia coli O157:H7 in milk based on catalase-mediated fluorescence quenching of CdTe quantum dots.

    Science.gov (United States)

    Chen, Rui; Huang, Xiaolin; Li, Juan; Shan, Shan; Lai, Weihua; Xiong, Yonghua

    2016-12-01

    Immunoassay is a powerful tool for rapid detection of food borne pathogens in food safety monitoring. However, conventional immunoassay always suffers from low sensitivity when it employs enzyme-catalyzing chromogenic substrates to generate colored molecules as signal outputs. In the present study, we report a novel fluorescence immunoassay for the sensitive detection of E. coli O157:H7 through combination of the ultrahigh bioactivity of catalase to hydrogen peroxide (H 2 O 2 ) and H 2 O 2 -sensitive mercaptopropionic acid modified CdTe QDs (MPA-QDs) as a signal transduction. Various parameters, including the concentrations of anti-E. coli O157:H7 polyclonal antibody and biotinylated monoclonal antibody, the amounts of H 2 O 2 and streptavidin labeled catalase (CAT), the hydrolysis temperature and time of CAT to H 2 O 2 , as well as the incubation time between H 2 O 2 and MPA-QDs, were systematically investigated and optimized. With optimal conditions, the catalase-mediated fluorescence quenching immunoassay exhibits an excellent sensitivity for E. coli O157:H7 with a detection limit of 5 × 10 2  CFU/mL, which was approximately 140 times lower than that of horseradish peroxidase-based colorimetric immunoassay. The reliability of the proposed method was further evaluated using E. coli O157:H7 spiked milk samples. The average recoveries of E. coli O157:H7 concentrations from 1.18 × 10 3  CFU/mL to 1.18 × 10 6  CFU/mL were in the range of 65.88%-105.6%. In brief, the proposed immunoassay offers a great potential for rapid and sensitive detection of other pathogens in food quality control. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Micromotor-based lab-on-chip immunoassays

    Science.gov (United States)

    García, Miguel; Orozco, Jahir; Guix, Maria; Gao, Wei; Sattayasamitsathit, Sirilak; Escarpa, Alberto; Merkoçi, Arben; Wang, Joseph

    2013-01-01

    Here we describe the first example of using self-propelled antibody-functionalized synthetic catalytic microengines for capturing and transporting target proteins between the different reservoirs of a lab-on-a-chip (LOC) device. A new catalytic polymer/Ni/Pt microtube engine, containing carboxy moieties on its mixed poly(3,4-ethylenedioxythiophene) (PEDOT)/COOH-PEDOT polymeric outermost layer, is further functionalized with the antibody receptor to selectively recognize and capture the target protein. The new motor-based microchip immunoassay operations are carried out without any bulk fluid flow, replacing the common washing steps in antibody-based protein bioassays with the active transport of the captured protein throughout the different reservoirs, where each step of the immunoassay takes place. A first microchip format involving an `on-the-fly' double-antibody sandwich assay (DASA) is used for demonstrating the selective capture of the target protein, in the presence of excess of non-target proteins. A secondary antibody tagged with a polymeric-sphere tracer allows the direct visualization of the binding events. In a second approach the immuno-nanomotor captures and transports the microsphere-tagged antigen through a microchannel network. An anti-protein-A modified microengine is finally used to demonstrate the selective capture, transport and convenient label-free optical detection of a Staphylococcus aureus target bacteria (containing proteinA in its cell wall) in the presence of a large excess of non-target (Saccharomyces cerevisiae) cells. The resulting nanomotor-based microchip immunoassay offers considerable potential for diverse applications in clinical diagnostics, environmental and security monitoring fields.Here we describe the first example of using self-propelled antibody-functionalized synthetic catalytic microengines for capturing and transporting target proteins between the different reservoirs of a lab-on-a-chip (LOC) device. A new catalytic

  13. Chimeric recombinant antibody fragments in cardiac troponin I immunoassay.

    Science.gov (United States)

    Hyytiä, Heidi; Heikkilä, Taina; Brockmann, Eeva-Christine; Kekki, Henna; Hedberg, Pirjo; Puolakanaho, Tarja; Lövgren, Timo; Pettersson, Kim

    2015-03-01

    To introduce a novel nanoparticle-based immunoassay for cardiac troponin I (cTnI) utilizing chimeric antibody fragments and to demonstrate that removal of antibody Fc-part and antibody chimerization decrease matrix related interferences. A sandwich-type immunoassay for cTnI based on recombinant chimeric (mouse variable/human constant) antigen binding (cFab) antibodies and intrinsically fluorescent nanoparticles was developed. To test whether using chimeric antibody fragments helps to avoid matrix related interferences, samples (n=39) with known amounts of triglycerides, bilirubin, rheumatoid factor (RF) or human anti-mouse antibodies (HAMAs) were measured with the novel assay, along with a previously published nanoparticle-based research assay with the same antibody epitopes. The limit of detection (LoD) was 3.30ng/L. Within-laboratory precision for 29ng/L and 2819ng/L cTnI were 13.7% and 15.9%, respectively. Regression analysis with Siemens ADVIA Centaur® yielded a slope (95% confidence intervals) of 0.18 (0.17-1.19) and a y-intercept of 1.94 (-1.28-3.91) ng/L. When compared to a previously published nanoparticle-based assay, the novel assay showed substantially reduced interference in the tested interference prone samples, 15.4 vs. 51.3%. A rheumatoid factor containing sample was decreased from 241ng/L to immunoassay for the detection of cTnI and decreased matrix related interferences, thus resulting in a lower number of falsely elevated cTnI-values. Copyright © 2014 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  14. iQuant™ Analyser: A rapid quantitative immunoassay reader.

    Science.gov (United States)

    Joseph, Jayaraj; Vasan, Jayaraman Kiruthi; Shah, Malay; Sivaprakasam, Mohansankar; Mahajan, Lalit

    2017-07-01

    Lateral flow immunoassays (LFIA) used in rapid quantitative point of care testing require an accurate, reliable and easy to operate instrument to read the LFIA kit and calculate the quantitative result value. We present iQuant® Analyser, an immunoassay reader designed for reading the Quanti® range of LFIA test kits for key markers such as HbA1C, Vitamin D, TSH etc. The instrument utilizes a laser based confocal optics system to capture the test and control lines from the LFIA kit, digitizes the fluorescent signal with high spatiotemporal resolution, computes necessary peak area ratios, applies calibration curves and declares the final result in an automated manner with minimal operator input. The instrument uses kit specific calibration information embedded on each LFIA test kit, to compute the final clinical parameter without using any external calibration chip. An intuitive icon based interface enables easy operation with minimal key presses, suited for point of care applications. The technology is designed in a modular manner to enable the instrument to perform tests on various parameters such as HbA1C, TSH, and Vitamin D etc without any hardware changes, using test-specific LFIA kits. The functional performance of the iQuant Analyser was verified over the range of expected area ratio values with standard reference cartridges that provided stable fluorescent lines. Repeatability of the instrument was found to be excellent with coefficient of variation (CoV) of area ratios found to be less than 1%. The inter-instrument reproducibility was also found to be good with CoV less than 4 %. Tests using blood samples with Quanti LFIA kits verified the accuracy of HbA1C results to be acceptable as per international standards with errors <; 4 %. The iQuant Analyser is a portable, easy to use rapid quantitative immunoassay reader best suited for point of care applications.

  15. Analytical evaluation of the novel Lumipulse G BRAHMS procalcitonin immunoassay

    OpenAIRE

    Ruzzenente, Orazio; Salvagno, Gian Luca; Gelati, Matteo; Lippi, Giuseppe

    2016-01-01

    Objectives: This study was designed to evaluate the analytical performance of the novel Lumipulse G1200 BRAHMS procalcitonin (PCT) immunoassay. Design and methods: This analytical evaluation encompassed the calculation of the limit of blank (LOB), limit of detection (LOD), functional sensitivity, intra- and inter-assay imprecision, confirmation of linearity and a comparison with the Vidas BRAHMS PCT assay. Results: The LOB, LOD and functional sensitivity were 0.0010 ng/mL, 0.0016 ng/mL and ...

  16. Tumor specific lung cancer diagnostics with multiplexed FRET immunoassays

    Science.gov (United States)

    Geißler, D.; Hill, D.; Löhmannsröben, H.-G.; Thomas, E.; Lavigne, A.; Darbouret, B.; Bois, E.; Charbonnière, L. J.; Ziessel, R. F.; Hildebrandt, N.

    2010-02-01

    An optical multiplexed homogeneous (liquid phase) immunoassay based on FRET from a terbium complex to eight different fluorescent dyes is presented. We achieved highly sensitive parallel detection of four different lung cancer specific tumor markers (CEA, NSE, SCC and CYFRA21-1) within a single assay and show a proof-of-principle for 5- fold multiplexing. The method is well suited for fast and low-cost miniaturized point-of-care testing as well as for highthroughput screening in a broad range of in-vitro diagnostic applications.

  17. Recalibration of the limiting antigen avidity EIA to determine mean duration of recent infection in divergent HIV-1 subtypes.

    Directory of Open Access Journals (Sweden)

    Yen T Duong

    Full Text Available Mean duration of recent infection (MDRI and misclassification of long-term HIV-1 infections, as proportion false recent (PFR, are critical parameters for laboratory-based assays for estimating HIV-1 incidence. Recent review of the data by us and others indicated that MDRI of LAg-Avidity EIA estimated previously required recalibration. We present here results of recalibration efforts using >250 seroconversion panels and multiple statistical methods to ensure accuracy and consensus.A total of 2737 longitudinal specimens collected from 259 seroconverting individuals infected with diverse HIV-1 subtypes were tested with the LAg-Avidity EIA as previously described. Data were analyzed for determination of MDRI at ODn cutoffs of 1.0 to 2.0 using 7 statistical approaches and sub-analyzed by HIV-1 subtypes. In addition, 3740 specimens from individuals with infection >1 year, including 488 from patients with AIDS, were tested for PFR at varying cutoffs.Using different statistical methods, MDRI values ranged from 88-94 days at cutoff ODn = 1.0 to 177-183 days at ODn = 2.0. The MDRI values were similar by different methods suggesting coherence of different approaches. Testing for misclassification among long-term infections indicated that overall PFRs were 0.6% to 2.5% at increasing cutoffs of 1.0 to 2.0, respectively. Balancing the need for a longer MDRI and smaller PFR (<2.0% suggests that a cutoff ODn = 1.5, corresponding to an MDRI of 130 days should be used for cross-sectional application. The MDRI varied among subtypes from 109 days (subtype A&D to 152 days (subtype C.Based on the new data and revised analysis, we recommend an ODn cutoff = 1.5 to classify recent and long-term infections, corresponding to an MDRI of 130 days (118-142. Determination of revised parameters for estimation of HIV-1 incidence should facilitate application of the LAg-Avidity EIA for worldwide use.

  18. Recalibration of the limiting antigen avidity EIA to determine mean duration of recent infection in divergent HIV-1 subtypes.

    Science.gov (United States)

    Duong, Yen T; Kassanjee, Reshma; Welte, Alex; Morgan, Meade; De, Anindya; Dobbs, Trudy; Rottinghaus, Erin; Nkengasong, John; Curlin, Marcel E; Kittinunvorakoon, Chonticha; Raengsakulrach, Boonyos; Martin, Michael; Choopanya, Kachit; Vanichseni, Suphak; Jiang, Yan; Qiu, Maofeng; Yu, Haiying; Hao, Yan; Shah, Neha; Le, Linh-Vi; Kim, Andrea A; Nguyen, Tuan Anh; Ampofo, William; Parekh, Bharat S

    2015-01-01

    Mean duration of recent infection (MDRI) and misclassification of long-term HIV-1 infections, as proportion false recent (PFR), are critical parameters for laboratory-based assays for estimating HIV-1 incidence. Recent review of the data by us and others indicated that MDRI of LAg-Avidity EIA estimated previously required recalibration. We present here results of recalibration efforts using >250 seroconversion panels and multiple statistical methods to ensure accuracy and consensus. A total of 2737 longitudinal specimens collected from 259 seroconverting individuals infected with diverse HIV-1 subtypes were tested with the LAg-Avidity EIA as previously described. Data were analyzed for determination of MDRI at ODn cutoffs of 1.0 to 2.0 using 7 statistical approaches and sub-analyzed by HIV-1 subtypes. In addition, 3740 specimens from individuals with infection >1 year, including 488 from patients with AIDS, were tested for PFR at varying cutoffs. Using different statistical methods, MDRI values ranged from 88-94 days at cutoff ODn = 1.0 to 177-183 days at ODn = 2.0. The MDRI values were similar by different methods suggesting coherence of different approaches. Testing for misclassification among long-term infections indicated that overall PFRs were 0.6% to 2.5% at increasing cutoffs of 1.0 to 2.0, respectively. Balancing the need for a longer MDRI and smaller PFR (<2.0%) suggests that a cutoff ODn = 1.5, corresponding to an MDRI of 130 days should be used for cross-sectional application. The MDRI varied among subtypes from 109 days (subtype A&D) to 152 days (subtype C). Based on the new data and revised analysis, we recommend an ODn cutoff = 1.5 to classify recent and long-term infections, corresponding to an MDRI of 130 days (118-142). Determination of revised parameters for estimation of HIV-1 incidence should facilitate application of the LAg-Avidity EIA for worldwide use.

  19. Evaluation of the Double Agar Gel Immunodiffusion Test and of the Enzyme-Linked Immunosorbent Assay in the Diagnosis and Follow-Up of Patients with Chronic Pulmonary Aspergillosis.

    Directory of Open Access Journals (Sweden)

    Priscila Zacarias de Azevedo

    Full Text Available The diagnosis of chronic pulmonary aspergillosis (CPA depends on the radiologic image and the identification of specific antibodies. The present study aimed to evaluate accuracy parameters of enzyme-linked immunosorbent assay (ELISA and of the determination of serum galactomannan level in the diagnosis of patients with CPA, comparing these results with the double agar gel immunodiffusion (DID test. In addition, the prevalence of cross-reactivity and the serological progression after treatment were evaluated by comparing DID and ELISA. Six study groups were formed: G1: 22 patients with CPA, 17 of whom had Aspergillus fungus ball, one chronic cavitary pulmonary aspergillosis (CCPA and four chronic fibrosing pulmonary aspergillosis (CFPA; G2: 28 patients with pulmonary tuberculosis (TB; G3: 23 patients with histoplasmosis (HST; G4: 50 patients with paracoccidioidomycosis (PCM; G5: 20 patients with cryptococcosis (CRC; and G6: 200 healthy controls. Serum antibodies were measured by DID and ELISA, with two antigen preparations--Aspergillus fumigatus (DID1, ELISA1 and a pool of A. fumigatus, A. flavus and A. niger antigens (DID2, ELISA2. The Platélia Aspergillus Enzyme Immunoassay (EIA kit was used to measure galactomannan. The cut-off points of ELISA were determined for each antigen preparation and for the 95% and 99% confidence intervals. Despite the low sensitivity, DID was the technique of choice due to its specificity, positive and negative predictive values and positive likelihood ratio-especially with the antigen pool and due to the low frequency of cross-reactivity. ELISA1 and a 0.090 cut-off showed high sensitivity, specificity and negative predictive value, but a high frequency of cross-reactivity with CRC. The best degree of agreement was observed between ELISA1 and ELISA2. The detection of serum galactomannan showed high sensitivity, comparable to ELISA2. The immunodiffusion test showed an excellent relationship with the progression after

  20. Inhibitors of endopeptidase and angiotensin-converting enzyme lead to an amplification of the morphological changes and an upregulation of the substance P system in a muscle overuse model.

    Science.gov (United States)

    Song, Yafeng; Stål, Per S; Yu, Ji-Guo; Lorentzon, Ronny; Backman, Clas; Forsgren, Sture

    2014-04-11

    We have previously observed, in studies on an experimental overuse model, that the tachykinin system may be involved in the processes of muscle inflammation (myositis) and other muscle tissue alterations. To further evaluate the significance of tachykinins in these processes, we have used inhibitors of neutral endopeptidase (NEP) and angiotensin-converting enzyme (ACE), substances which are known to terminate the activity of various endogenously produced substances, including tachykinins. Injections of inhibitors of NEP and ACE, as well as the tachykinin substance P (SP), were given locally outside the tendon of the triceps surae muscle of rabbits subjected to marked overuse of this muscle. A control group was given NaCl injections. Evaluations were made at 1 week, a timepoint of overuse when only mild inflammation and limited changes in the muscle structure are noted in animals not treated with inhibitors. Both the soleus and gastrocnemius muscles were examined morphologically and with immunohistochemistry and enzyme immunoassay (EIA). A pronounced inflammation (myositis) and changes in the muscle fiber morphology, including muscle fiber necrosis, occurred in the overused muscles of animals given NEP and ACE inhibitors. The morphological changes were clearly more prominent than for animals subjected to overuse and NaCl injections (NaCl group). A marked SP-like expression, as well as a marked expression of the neurokinin-1 receptor (NK-1R) was found in the affected muscle tissue in response to injections of NEP and ACE inhibitors. The concentration of SP in the muscles was also higher than that for the NaCl group. The observations show that the local injections of NEP and ACE inhibitors led to marked SP-like and NK-1R immunoreactions, increased SP concentrations, and an amplification of the morphological changes in the tissue. The injections of the inhibitors thus led to a more marked myositis process and an upregulation of the SP system. Endogenously produced

  1. KIMS, CEDIA, and HS-CEDIA immunoassays are inadequately sensitive for detection of benzodiazepines in urine from patients treated for chronic pain.

    Science.gov (United States)

    Darragh, Alicia; Snyder, Marion L; Ptolemy, Adam S; Melanson, Stacy

    2014-01-01

    Patients treated for chronic pain may frequently undergo urine drug testing to monitor medication compliance and detect undisclosed prescribed or illicit drug use. Due to the increasing use and abuse of benzodiazepines, this class of medications is often included in drug screening panels. However, immunoassay-based methods lack the requisite sensitivity for detecting benzodiazepine use in this population primarily due to their poor cross-reactivity with several major urinary benzodiazepine metabolites. A High Sensitivity Cloned Enzyme Donor Immunoassay (HS-CEDIA), in which beta-glucuronidase is added to the reagent, has been shown to perform better than traditional assays, but its performance in patients treated for chronic pain is not well characterized. To determine the diagnostic accuracy of HS-CEDIA, as compared to the Cloned Enzyme Donor Immunoassay (CEDIA) and Kinetic Interaction of Microparticles in Solution (KIMS) screening immunoassays and liquid chromatography-tandem mass spectrometry (LC-MS/MS), for monitoring benzodiazepine use in patients treated for chronic pain. A study of the diagnostic accuracy of urine benzodiazepine immunoassays. The study was conducted at an academic tertiary care hospital with a clinical laboratory that performs urine drug testing for monitoring medication compliance in pain management. A total of 299 urine specimens from patients treated for chronic pain were screened for the presence of benzodiazepines using the HS-CEDIA, CEDIA, and KIMS assays. The sensitivity and specificity of the screening assays were determined using the LC-MS/MS results as the reference method. Of the 299 urine specimens tested, 141 (47%) confirmed positive for one or more of the benzodiazepines/metabolites by LC-MS/MS. All 3 screens were 100% specific with no false-positive results. The CEDIA and KIMS sensitivities were 55% (78/141) and 47% (66/141), respectively. Despite the relatively higher sensitivity of the HS-CEDIA screening assay (78%; 110

  2. Production of monoclonal antibodies for sandwich immunoassay detection of Pacific ciguatoxins.

    Science.gov (United States)

    Tsumuraya, Takeshi; Fujii, Ikuo; Hirama, Masahiro

    2010-10-01

    Ciguatoxins are the major causative toxins of ciguatera seafood poisoning. Limited availability of ciguatoxins has hampered the development of a reliable and specific immunoassay for detecting these toxins in contaminated fish. Monoclonal antibodies (mAbs) specific against both ends of Pacific ciguatoxins CTX3C and 51-hydroxyCTX3C were prepared by immunization of mice with the protein conjugates of rationally designed synthetic haptens in place of the natural toxin. Haptenic groups that possess a surface area larger than 400 A(2) were required to produce mAbs that can bind strongly to CTX3C or 51-hydroxyCTX3C. A direct sandwich enzyme-linked immunosorbent assay (ELISA) using these mAbs was established to detect CTX3C and 51-hydroxyCTX3C at the ppb level with no cross-reactivity against the other marine toxins, including brevetoxin A, brevetoxin B, okadaic acid, or maitotoxin. Copyright 2009 Elsevier Ltd. All rights reserved.

  3. A multiplexed immunoassay for detection of antibodies to Actinobacillus pleuropneumoniae (App) in pigs

    DEFF Research Database (Denmark)

    Berger, Sanne Schou; Boas, Ulrik; Andresen, Lars Ole

    2014-01-01

    our diagnostic tools, we are currently developing a novel indirect fluorescent microsphere immunoassay that can facilitate simultaneous detection of antibodies towards multiple App serovars within a single serum sample volume. The multiplex immunoassay is based on Luminex technology (8) and has...

  4. Effective detection of toxigenic Clostridium difficile by a two-step algorithm including tests for antigen and cytotoxin.

    Science.gov (United States)

    Ticehurst, John R; Aird, Deborah Z; Dam, Lisa M; Borek, Anita P; Hargrove, John T; Carroll, Karen C

    2006-03-01

    We evaluated a two-step algorithm for detecting toxigenic Clostridium difficile: an enzyme immunoassay for glutamate dehydrogenase antigen (Ag-EIA) and then, for antigen-positive specimens, a concurrent cell culture cytotoxicity neutralization assay (CCNA). Antigen-negative results were > or = 99% predictive of CCNA negativity. Because the Ag-EIA reduced cell culture workload by approximately 75 to 80% and two-step testing was complete in CCNA alone had been performed on all 5,887 specimens.

  5. Enzyme detection by microfluidics

    DEFF Research Database (Denmark)

    2013-01-01

    Microfluidic-implemented methods of detecting an enzyme, in particular a DNA-modifying enzyme, are provided, as well as methods for detecting a cell, or a microorganism expressing said enzyme. The enzyme is detected by providing a nucleic acid substrate, which is specifically targeted...... by that enzyme...

  6. Enhanced Fluorescent Immunoassays on Silver Fractal-like Structures

    Science.gov (United States)

    Shtoyko, Tanya; Matveeva, Evgenia G.; Chang, I-Fen; Gryczynski, Zygmunt; Goldys, Ewa; Gryczynski, Ignacy

    2009-01-01

    Using the effect of the fluorescence enhancement in close proximity to metal nanostructures, we have been able to demonstrate ultrasensitive immunoassays suitable for the detection of biomarkers. Silver fractal-like structures have been grown by electrochemical reduction of silver on the surface of glass slides. A model immunoassay was performed on the slide surface with rabbit IgG (antigen) non-covalently immobilized on the slide, and Rhodamine Red-X labeled anti-rabbit IgG conjugate subsequently bound to the immobilized antigen. The fluorescence signal was measured from the glass-fractals surface using a confocal microscope, and the images were compared to the images from the same surface not coated with fractals. Our results showed significant enhancement (more than 100-fold) of the signal detected on fractals compared to bare glass. We thus demonstrate that such fractal-like structures can assist in improving the signals from assays used in medical diagnostics, especially those for analytes with molecular weight under 100 kD. PMID:18288816

  7. Trends in interfacial design for surface plasmon resonance based immunoassays

    Energy Technology Data Exchange (ETDEWEB)

    Shankaran, Dhesingh Ravi [Art, Science and Technology Center for Cooperative Research, Kyushu University, Kasuga-shi, Fukuoka, 816-8580 (Japan); Miura, Norio [Art, Science and Technology Center for Cooperative Research, Kyushu University, Kasuga-shi, Fukuoka, 816-8580 (Japan)

    2007-12-07

    Immunosensors based on surface plasmon resonance (SPR) have become a promising tool in sensor technology for biomedical, food, environmental, industrial and homeland security applications. SPR is a surface sensitive optical technique, suitable for real-time and label-free analysis of biorecognition events at functional transducer surfaces. Fabrication of highly active and robust sensing surfaces is an important part in immunoassays because the quality, quantity, chemistry and topography of the interfacial biomembranes play a major role in immunosensor performance. Eventually, a variety of immobilization methods such as physical adsorption, covalent coupling, Langmuir-Blodgett film, polymer thin film, self-assembly, sol-gel, etc, have been introduced over the years for the immobilization of biomolecules (antibody or antigen) on the transducer surfaces. The selection of an immobilization method for an immunoassay is governed by several factors such as nature and stability of the biomolecules, target analyte, application, detection principle, mode of signal transduction, matrix complexity, etc. This paper provides an overview of the various surface modification methods for SPR based immunosensor fabrication. The preparation, structure and application of different functional interfacial surfaces have been discussed along with a brief introduction to the SPR technology, biomolecules and detection principles. (review article)

  8. Trends in interfacial design for surface plasmon resonance based immunoassays

    International Nuclear Information System (INIS)

    Shankaran, Dhesingh Ravi; Miura, Norio

    2007-01-01

    Immunosensors based on surface plasmon resonance (SPR) have become a promising tool in sensor technology for biomedical, food, environmental, industrial and homeland security applications. SPR is a surface sensitive optical technique, suitable for real-time and label-free analysis of biorecognition events at functional transducer surfaces. Fabrication of highly active and robust sensing surfaces is an important part in immunoassays because the quality, quantity, chemistry and topography of the interfacial biomembranes play a major role in immunosensor performance. Eventually, a variety of immobilization methods such as physical adsorption, covalent coupling, Langmuir-Blodgett film, polymer thin film, self-assembly, sol-gel, etc, have been introduced over the years for the immobilization of biomolecules (antibody or antigen) on the transducer surfaces. The selection of an immobilization method for an immunoassay is governed by several factors such as nature and stability of the biomolecules, target analyte, application, detection principle, mode of signal transduction, matrix complexity, etc. This paper provides an overview of the various surface modification methods for SPR based immunosensor fabrication. The preparation, structure and application of different functional interfacial surfaces have been discussed along with a brief introduction to the SPR technology, biomolecules and detection principles. (review article)

  9. Species Specific Bacterial Spore Detection Using Lateral-Flow Immunoassay with DPA-Triggered Tb Luminescence

    Science.gov (United States)

    Ponce, Adrian

    2003-01-01

    A method of detecting bacterial spores incorporates (1) A method of lateral-flow immunoassay in combination with (2) A method based on the luminescence of Tb3+ ions to which molecules of dipicolinic acid (DPA) released from the spores have become bound. The present combination of lateral-flow immunoassay and DPA-triggered Tb luminescence was developed as a superior alternative to a prior lateral-flow immunoassay method in which detection involves the visual observation and/or measurement of red light scattered from colloidal gold nanoparticles. The advantage of the present combination method is that it affords both (1) High selectivity for spores of the species of bacteria that one seeks to detect (a characteristic of lateral-flow immunoassay in general) and (2) Detection sensitivity much greater (by virtue of the use of DPA-triggered Tb luminescence instead of gold nanoparticles) than that of the prior lateral-flow immunoassay method

  10. Novel strategies to enhance lateral flow immunoassay sensitivity for detecting foodborne pathogens.

    Science.gov (United States)

    Shan, Shan; Lai, Weihua; Xiong, Yonghua; Wei, Hua; Xu, Hengyi

    2015-01-28

    Food contaminated by foodborne pathogens causes diseases, affects individuals, and even kills those affected individuals. As such, rapid and sensitive detection methods should be developed to screen pathogens in food. One current detection method is lateral flow immunoassay, an efficient technique because of several advantages, including rapidity, simplicity, stability, portability, and sensitivity. This review presents the format and principle of lateral flow immunoassay strip and the development of conventional lateral flow immunoassay for detecting foodborne pathogens. Furthermore, novel strategies that can be applied to enhance the sensitivity of lateral flow immunoassay to detect foodborne pathogens are presented; these strategies include innovating new label application, designing new formats of lateral flow immunoassay, combining with other methods, and developing signal amplification systems. With these advancements, detection sensitivity and detection time can be greatly improved.

  11. Redox-Magnetohydrodynamic Microfluidics Without Channels and Compatible with Electrochemical Detection Under Immunoassay Conditions

    Science.gov (United States)

    Weston, Melissa C.; Nash, Christena K.; Fritsch, Ingrid

    2010-01-01

    A unique capability of redox-magnetohydrodynamics (redox-MHD) for handling liquids on a small scale was demonstrated. A 1.2-μL solution plug was pumped from an injection site to a detector without the need for a channel to direct the flow. The redox pumping species did not interfere with enzymatic activity in a solution compatible with enzyme-linked immunoassays. Alkaline phosphatase (AP), a common enzyme label, converted p-aminophenyl phosphate (PAPP) to p-aminophenol (PAPR) in the presence of 2.5 mM Ru(NH3)6Cl2 and 2.5 mM Ru(NH3)6 Cl3, in 0.1 M Tris buffer (pH=9). A solution plug containing PAPP (no AP) was pumped through the surrounding solution containing AP (no PAPP), and the enzymatically-generated PAPR was easily detected and distinguishable electrochemically from the pumping species with square wave voltammetry down to 0.1 mM concentrations. The test device consisted of a silicon chip containing individually-addressable microband electrodes, placed on a 0.5-T NdFeB permanent magnet with the field oriented perpendicular to the chip. A 8.0-mm wide × 15.5-mm long × 1.5-mm high volume of solution was contained by a poly(dimethylsiloxane) gasket and capped with a glass slide. A steady-state fluid velocity of ~30 μm/s was generated in a reinforcing flow configuration between oppositely polarized sets of pumping electrodes with ~2.1 μA. PMID:20681513

  12. The interplay between research, research-based advice and the development of the Greenlandic Governmental environmental legislation and practices in respect to EIA and SEA

    DEFF Research Database (Denmark)

    Mosbech, Anders

    Government decides to grant licensees for oil exploration the operators have to develop specific EIAs for all major activities. The activities shall be based on BEP (Best Environmental Practice) and BAT (Best Environmental Technology) principles, and operators shall conduct supplementary environmental...

  13. Relation of Environmental Impact Assessment (EIA and Strategic Environmental Assessment (SEA and the Importance of Strategic Environmental Assessment in Landscape Planning Process

    Directory of Open Access Journals (Sweden)

    Gizem CENGİZ GÖKÇE

    2015-07-01

    Full Text Available The main goal in the countries which have not completed their development progress is industrialization and development just as soon as possible. Therefore, negative effects of industrialization and development on envi ronment and/or nature cannot be mostly discussed adequately. One of the planning approach instruments that targets sustainability, Environmental Impact Assessment (EIA is used in many countries effectively. But in recent years, that has understood; EIA is an impact assessment instrument that contains defensive preventions only on the basis of projects and this situation has caused some concerns against EIA. In this direction, Strategical Environmental Assessment (SEA exists as the final point of the instruments which are formed to provide sustainable development . In this study; the importance and the requirement of effectively taking a role of landscape architectures that have ecological based job, in the SEA workings which isn’t have got a legal status in Turkey yet, are emphasized by reviewing the relations between EIA and SEA concepts.

  14. Application of an enzyme-linked immunosorbent assay (ELISA) to determine paraquat residues in milk, beef, and potatoes

    International Nuclear Information System (INIS)

    Van Emon, J.; Seiber, J.; Hammock, B.

    1987-01-01

    The USDA Food Safety and Inspection Service (FSIS) has included paraquat on its list of compounds to be considered for monitoring in foods. However, present methods do not easily accommodate the processing of large numbers of samples, thus limiting routine monitoring of the compound. The conventional method, based on spectrophotometry of reduced paraquat solutions, requires time-consuming sample preparation. Although the advantages of immunoassays for pesticide residue analysis have been pointed out, the reported immunoassays for paraquat have only been applied to cases of clinical poisoning or human exposure assessment. In this study, spiked milk, potato, and beef were analyzed directly, without prior cleanup, by an enzyme-linked immunosorbent assay (ELISA)

  15. Comparison between the two-step and the three-step algorithms for the detection of toxigenic Clostridium difficile.

    Science.gov (United States)

    Qutub, M O; AlBaz, N; Hawken, P; Anoos, A

    2011-01-01

    To evaluate usefulness of applying either the two-step algorithm (Ag-EIAs and CCNA) or the three-step algorithm (all three assays) for better confirmation of toxigenic Clostridium difficile. The antigen enzyme immunoassays (Ag-EIAs) can accurately identify the glutamate dehydrogenase antigen of toxigenic and nontoxigenic Clostridium difficile. Therefore, it is used in combination with a toxin-detecting assay [cell line culture neutralization assay (CCNA), or the enzyme immunoassays for toxins A and B (TOX-A/BII EIA)] to provide specific evidence of Clostridium difficile-associated diarrhoea. A total of 151 nonformed stool specimens were tested by Ag-EIAs, TOX-A/BII EIA, and CCNA. All tests were performed according to the manufacturer's instructions and the results of Ag-EIAs and TOX-A/BII EIA were read using a spectrophotometer at a wavelength of 450 nm. A total of 61 (40.7%), 38 (25.3%), and 52 (34.7%) specimens tested positive with Ag-EIA, TOX-A/BII EIA, and CCNA, respectively. Overall, the sensitivity, specificity, negative predictive value, and positive predictive value for Ag-EIA were 94%, 87%, 96.6%, and 80.3%, respectively. Whereas for TOX-A/BII EIA, the sensitivity, specificity, negative predictive value, and positive predictive value were 73.1%, 100%, 87.5%, and 100%, respectively. With the two-step algorithm, all 61 Ag-EIAs-positive cases required 2 days for confirmation. With the three-step algorithm, 37 (60.7%) cases were reported immediately, and the remaining 24 (39.3%) required further testing by CCNA. By applying the two-step algorithm, the workload and cost could be reduced by 28.2% compared with the three-step algorithm. The two-step algorithm is the most practical for accurately detecting toxigenic Clostridium difficile, but it is time-consuming.

  16. Comparison between the two-step and the three-step algorithms for the detection of toxigenic Clostridium difficile

    Directory of Open Access Journals (Sweden)

    M O Qutub

    2011-01-01

    Full Text Available Purpose: To evaluate usefulness of applying either the two-step algorithm (Ag-EIAs and CCNA or the three-step algorithm (all three assays for better confirmation of toxigenic Clostridium difficile. The antigen enzyme immunoassays (Ag-EIAs can accurately identify the glutamate dehydrogenase antigen of toxigenic and nontoxigenic Clostridium difficile. Therefore, it is used in combination with a toxin-detecting assay [cell line culture neutralization assay (CCNA, or the enzyme immunoassays for toxins A and B (TOX-A/BII EIA] to provide specific evidence of Clostridium difficile-associated diarrhoea. Materials and Methods: A total of 151 nonformed stool specimens were tested by Ag-EIAs, TOX-A/BII EIA, and CCNA. All tests were performed according to the manufacturer′s instructions and the results of Ag-EIAs and TOX-A/BII EIA were read using a spectrophotometer at a wavelength of 450 nm. Results: A total of 61 (40.7%, 38 (25.3%, and 52 (34.7% specimens tested positive with Ag-EIA, TOX-A/BII EIA, and CCNA, respectively. Overall, the sensitivity, specificity, negative predictive value, and positive predictive value for Ag-EIA were 94%, 87%, 96.6%, and 80.3%, respectively. Whereas for TOX-A/BII EIA, the sensitivity, specificity, negative predictive value, and positive predictive value were 73.1%, 100%, 87.5%, and 100%, respectively. With the two-step algorithm, all 61 Ag-EIAs-positive cases required 2 days for confirmation. With the three-step algorithm, 37 (60.7% cases were reported immediately, and the remaining 24 (39.3% required further testing by CCNA. By applying the two-step algorithm, the workload and cost could be reduced by 28.2% compared with the three-step algorithm. Conclusions: The two-step algorithm is the most practical for accurately detecting toxigenic Clostridium difficile, but it is time-consuming.

  17. Elevated Liver Enzymes

    Science.gov (United States)

    Symptoms Elevated liver enzymes By Mayo Clinic Staff Elevated liver enzymes may indicate inflammation or damage to cells in the liver. Inflamed or ... than normal amounts of certain chemicals, including liver enzymes, into the bloodstream, which can result in elevated ...

  18. Evaluation of envelope glycoprotein E(rns) of an atypical bovine pestivirus as antigen in a microsphere immunoassay for the detection of antibodies against bovine viral diarrhea virus 1 and atypical bovine pestivirus.

    Science.gov (United States)

    Vijayaraghavan, Balaje; Xia, Hongyan; Harimoorthy, Rajiv; Liu, Lihong; Belák, Sándor

    2012-11-01

    Atypical bovine pestiviruses are related antigenically and phylogenetically to bovine viral diarrhea viruses (BVDV-1 and BVDV-2), and may cause the same clinical manifestations in animals. Glycoprotein E(rns) of an atypical bovine pestivirus Th/04_KhonKaen was produced in a baculovirus expression system and was purified by affinity chromatography. The recombinant E(rns) protein was used as an antigen in a microsphere immunoassay for the detection of antibodies against BVDV-1 and atypical bovine pestivirus. The diagnostic performance of the new method was evaluated by testing a total of 596 serum samples, and the assay was compared with enzyme-linked immunosorbent assay (ELISA). Based on the negative/positive cut-off median fluorescence intensity (MFI) value of 2800, the microsphere immunoassay had a sensitivity of 100% and specificity of 100% compared to ELISA. The immunoassay was able to detect antibodies against both BVDV-1 and the atypical pestivirus. This novel microsphere immunoassay has the potential to be multiplexed for simultaneous detection of antibodies against different bovine pathogens in a high-throughput and economical way. Copyright © 2012 Elsevier B.V. All rights reserved.

  19. Color encoded microbeads-based flow cytometric immunoassay for polycyclic aromatic hydrocarbons in food

    International Nuclear Information System (INIS)

    Meimaridou, Anastasia; Haasnoot, Willem; Noteboom, Linda; Mintzas, Dimitrios; Pulkrabova, Jana; Hajslova, Jana; Nielen, Michel W.F.

    2010-01-01

    Food contamination caused by chemical hazards such as persistent organic pollutants (POPs) is a worldwide public health concern and requires continuous monitoring. The chromatography-based analysis methods for POPs are accurate and quite sensitive but they are time-consuming, laborious and expensive. Thus, there is a need for validated simplified screening tools, which are inexpensive, rapid, have automation potential and can detect multiple POPs simultaneously. In this study we developed a flow cytometry-based immunoassay (FCIA) using a color-encoded microbeads technology to detect benzo[a]pyrene (BaP) and other polycyclic aromatic hydrocarbons (PAHs) in buffer and food extracts as a starting point for the future development of rapid multiplex assays including other POPs in food, such as polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs). A highly sensitive assay for BaP was obtained with an IC 50 of 0.3 μg L -1 using a monoclonal antibody (Mab22F12) against BaP, similar to the IC 50 of a previously described enzyme-linked immunosorbent assay (ELISA) using the same Mab. Moreover, the FCIA was 8 times more sensitive for BaP compared to a surface plasmon resonance (SPR)-based biosensor immunoassay (BIA) using the same reagents. The selectivity of the FCIAs was tested, with two Mabs against BaP for 25 other PAHs, including two hydroxyl PAH metabolites. Apart from BaP, the FCIAs can detect PAHs such as indenol[1,2,3-cd]pyrene (IP), benz[a]anthracene (BaA), and chrysene (CHR) which are also appointed by the European Food Safety Authority (EFSA) as suitable indicators of PAH contamination in food. The FCIAs results were in agreement with those obtained with gas chromatography-mass spectrometry (GC-MS) for the detection of PAHs in real food samples of smoked carp and wheat flour and has great potential for the future routine application of this assay in a simplex or multiplex format in combination with simplified extraction procedure which are

  20. Lateral Flow Immunoassays for Ebola Virus Disease Detection in Liberia.

    Science.gov (United States)

    Phan, Jill C; Pettitt, James; George, Josiah S; Fakoli, Lawrence S; Taweh, Fahn M; Bateman, Stacey L; Bennett, Richard S; Norris, Sarah L; Spinnler, David A; Pimentel, Guillermo; Sahr, Phillip K; Bolay, Fatorma K; Schoepp, Randal J

    2016-10-15

     Lateral flow immunoassays (LFIs) are point-of-care diagnostic assays that are designed for single use outside a formal laboratory, with in-home pregnancy tests the best-known example of these tests. Although the LFI has some limitations over more-complex immunoassay procedures, such as reduced sensitivity and the potential for false-positive results when using complex sample matrices, the assay has the benefits of a rapid time to result and ease of use. These benefits make it an attractive option for obtaining rapid results in an austere environment. In an outbreak of any magnitude, a field-based rapid diagnostic assay would allow proper patient transport and for safe burials to be conducted without the delay caused by transport of samples between remote villages and testing facilities. Use of such point-of-care instruments in the ongoing Ebola virus disease (EVD) outbreak in West Africa would have distinct advantages in control and prevention of local outbreaks, but proper understanding of the technology and interpretation of results are important.  In this study, a LFI, originally developed by the Naval Medical Research Center for Ebola virus environmental testing, was evaluated for its ability to detect the virus in clinical samples in Liberia. Clinical blood and plasma samples and post mortem oral swabs submitted to the Liberian Institute for Biomedical Research, the National Public Health Reference Laboratory for EVD testing, were tested and compared to results of real-time reverse transcription-polymerase chain reaction (rRT-PCR), using assays targeting Ebola virus glycoprotein and nucleoprotein.  The LFI findings correlated well with those of the real-time RT-PCR assays used as benchmarks.  Rapid antigen-detection tests such as LFIs are attractive alternatives to traditional immunoassays but have reduced sensitivity and specificity, resulting in increases in false-positive and false-negative results. An understanding of the strengths, weaknesses

  1. Chemiluminescence immunoassay for free prostate-specific antigen

    International Nuclear Information System (INIS)

    Zhang Xuefeng; Liu Yibing; Xu Wenge; Li Ziying; Han Zhiquan; Liu Ting

    2008-01-01

    A sandwich chemiluminescence immunoassay (CLIA) for serum free prostate-specific antigen (F-PSA) was developed. One antibody against total PSA was coated on the micro-plate, the other antibody against F-PSA was labeled with horseradish peroxidase. The detection limit is established as 0.01 ng/mL (n=10, mean of zero standard + 2SD) and the intra-and inter-assay coefficient s of variation (CV) is in the range of 31 8%-51 4% and 10.8%-17.7%, respectively. Compared with Mono bind F-PSA CLIA kits, the correlative equation is y= 0.72x- 0.22, and r = 0.90. The standard range for the method is 0.2-10 ng/mL, and it presents good linearity. (authors)

  2. Clinical development of placental malaria vaccines and immunoassays harmonization

    DEFF Research Database (Denmark)

    Chêne, Arnaud; Houard, Sophie; Nielsen, Morten A

    2016-01-01

    Placental malaria caused by Plasmodium falciparum infection constitutes a major health problem manifesting as severe disease and anaemia in the mother, impaired fetal development, low birth weight or spontaneous abortion. Prevention of placental malaria currently relies on two key strategies...... that are losing efficacy due to spread of resistance: long-lasting insecticide-treated nets and intermittent preventive treatment during pregnancy. A placental malaria vaccine would be an attractive, cost-effective complement to the existing control tools. Two placental malaria vaccine candidates are currently...... in Phase Ia/b clinical trials. During two workshops hosted by the European Vaccine Initiative, one in Paris in April 2014 and the other in Brussels in November 2014, the main actors in placental malaria vaccine research discussed the harmonization of clinical development plans and of the immunoassays...

  3. Performance of immunoassay kits for site characterization and remediation

    International Nuclear Information System (INIS)

    Waters, L.C.; Palausky, A.; Counts, R.W.; Jenkins, R.A.

    1995-01-01

    The US Department of Energy (DOE) is supporting efforts to identify, validate and implement the use of effective, low-cost alternatives to currently used analytical methods for environmental management. As part of that program, we have evaluated the performances of a number of immunoassay (IA) kits with specificities for environmental contaminants of concern to the DOE. The studies were done in the laboratory using both spiked and field test samples. The analyte specificity and manufacturers of the kits evaluated were the following: mercury, BioNebraska; polychlorinated biphenyls (PCBs), EnSys and Millipore; petroleum fuel hydrocarbons, Millipore and Ohmicron; and polyaromatic hydrocarbons (PAHs), Ohmicron and Millipore. The kits were used in either a semiquantitative or quantitative format according to the preference of the manufacturers

  4. Biocatalytic Single Enzyme Nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Grate, Jay W.; Kim, Jungbae

    2004-03-31

    As an innovative way of enzyme stabilization, we recently developed a new enzyme composite of nano-meter scale that we call "single-enzyme nanoparticles (SENs)" (9). Each enzyme molecule is surrounded with a porous composite organic/inorganic network of less than a few nanometers think. This approach represents a new type of enzyme-containing nanostructure. In experiments with perotease (chymotrypsin, CT), the activity of single enzyme nanoparticle form of the enzyme was greatly stabilized compared to the free form, without imposing a serious mass transfer limitation of substrates. In this chapter we will describe the synthesis, characterization and catalytic activity of the new SENs.

  5. Development of Dual Quantitative Lateral Flow Immunoassay for the Detection of Mycotoxins.

    Science.gov (United States)

    Wang, Yuan-Kai; Yan, Ya-Xian; Sun, Jian-He

    2017-01-01

    Lateral flow immunoassays have been widely used in recent years for detection of toxins, heavy metals, and biomarkers. To improve the efficiency of individual lateral flow immunoassays, multiplex analytical strips play an important role in the detection of several important analytes. In this chapter, development of a dual lateral flow immunoassay is presented for detection of a variety of low molecular weight molecules. Various buffers, additives, and materials are introduced and evaluated. Depending on the analyte to be tested, the technique allows for selection of optimum buffers, additives, and other materials.

  6. Evaluation of an enzyme-linked immunosorbent assay for serological surveillance of infection with Actinobacillus pleuropneumoniae serotype 5 in pig herds

    DEFF Research Database (Denmark)

    Klausen, Joan; Andresen, Lars Ole; Barfod, Kristen

    2002-01-01

    An indirect enzyme-linked immunoassay for serological surveillance of infection of pigs with Actinobacillus pleuropneumoniae (Ap) serotype 5 was developed. The antigen used was prepared from Ap serotype 5b strain L20. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis...

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

    NARCIS (Netherlands)

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

    1998-01-01

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

  8. Ultrasensitive fluorescence immunoassay for detection of ochratoxin A using catalase-mediated fluorescence quenching of CdTe QDs

    Science.gov (United States)

    Huang, Xiaolin; Zhan, Shengnan; Xu, Hengyi; Meng, Xianwei; Xiong, Yonghua; Chen, Xiaoyuan

    2016-04-01

    Herein, for the first time we report an improved competitive fluorescent enzyme linked immunosorbent assay (ELISA) for the ultrasensitive detection of ochratoxin A (OTA) by using hydrogen peroxide (H2O2)-induced fluorescence quenching of mercaptopropionic acid-modified CdTe quantum dots (QDs). In this immunoassay, catalase (CAT) was labeled with OTA as a competitive antigen to connect the fluorescence signals of the QDs with the concentration of the target. Through the combinatorial use of H2O2-induced fluorescence quenching of CdTe QDs as a fluorescence signal output and the ultrahigh catalytic activity of CAT to H2O2, our proposed method could be used to perform a dynamic linear detection of OTA ranging from 0.05 pg mL-1 to 10 pg mL-1. The half maximal inhibitory concentration was 0.53 pg mL-1 and the limit of detection was 0.05 pg mL-1. These values were approximately 283- and 300-folds lower than those of horseradish peroxidase (HRP)-based conventional ELISA, respectively. The reported method is accurate, highly reproducible, and specific against other mycotoxins in agricultural products as well. In summary, the developed fluorescence immunoassay based on H2O2-induced fluorescence quenching of CdTe QDs can be used for the rapid and highly sensitive detection of mycotoxins or haptens in food safety monitoring.Herein, for the first time we report an improved competitive fluorescent enzyme linked immunosorbent assay (ELISA) for the ultrasensitive detection of ochratoxin A (OTA) by using hydrogen peroxide (H2O2)-induced fluorescence quenching of mercaptopropionic acid-modified CdTe quantum dots (QDs). In this immunoassay, catalase (CAT) was labeled with OTA as a competitive antigen to connect the fluorescence signals of the QDs with the concentration of the target. Through the combinatorial use of H2O2-induced fluorescence quenching of CdTe QDs as a fluorescence signal output and the ultrahigh catalytic activity of CAT to H2O2, our proposed method could be used to

  9. Development of a flow cytometric bead immunoassay and its assessment as a possible aid to potency evaluation of enterotoxaemia vaccines

    Directory of Open Access Journals (Sweden)

    Angela Buys

    2014-03-01

    Full Text Available Enterotoxaemia, an economically important disease of sheep, goats and calves, is caused by systemic effects of the epsilon toxin produced by the anaerobic bacterium Clostridium perfringens type D. The only practical means of controlling the occurrence of enterotoxaemia is to immunise animals by vaccination. The vaccine is prepared by deriving a toxoid from the bacterial culture filtrate and the potency of the vaccine is tested with the in vivo mouse neutralisation test (MNT. Due to ethical, economic and technical reasons, alternative in vitro assays are needed. In this study an indirect cytometric bead immunoassay (I-CBA was developed for use in vaccine potency testing and the results were compared with those obtained using an indirect enzyme-linked immunosorbent assay (I-ELISA and the MNT. Sera were collected from guinea pigs immunised with three different production batches of enterotoxaemia vaccine and the levels of anti-epsilon toxin antibodies were determined. Although the intra- and inter-assay variability was satisfactory, epsilon antitoxin levels determined by both the I-ELISA and indirect cytometric bead immunoassay (I-CBA tests were higher than those of the MNT assay. In contrast to the MNT, all of the serum samples were identified as having antitoxin levels above the required minimum (not less than 5 U/mL. These results indicate that the respective in vitro tests in their current formats are not yet suitable alternatives to the in vivo MNT. The growing demand for a more humane, cost-effective and efficient method for testing the potency of enterotoxaemia vaccines, however, provides a strong impetus for further optimisation and standardisation of the I-CBA assay but further analytical research is required.

  10. Sequential push-pull pumping mechanism for washing and evacuation of an immunoassay reaction chamber on a microfluidic CD platform.

    Directory of Open Access Journals (Sweden)

    Tzer Hwai Gilbert Thio

    Full Text Available A centrifugal compact disc (CD microfluidic platform with reservoirs, micro-channels, and valves can be employed for implementing a complete immunoassay. Detection or biosensor chambers are either coated for immuno-interaction or a biosensor chip is inserted in them. On microfluidic CDs featuring such multi-step chemical/biological processes, the biosensor chamber must be repeatedly filled with fluids such as enzymes solutions, buffers, and washing solutions. After each filling step, the biosensor chamber needs to be evacuated by a passive siphoning process to prepare it for the next step in the assay. However, rotational speed dependency and limited space on a CD are two big obstacles to performing such repetitive filling and siphoning steps. In this work, a unique thermo-pneumatic (TP Push-Pull pumping method is employed to provide a superior alternative biosensor chamber filling and evacuation technique. The proposed technique is demonstrated on two CD designs. The first design features a simple two-step microfluidic process to demonstrate the evacuation technique, while the second design shows the filling and evacuation technique with an example sequence for an actual immunoassay. In addition, the performance of the filling and evacuation technique as a washing step is also evaluated quantitatively and compared to the conventional manual bench top washing method. The two designs and the performance evaluation demonstrate that the technique is simple to implement, reliable, easy to control, and allows for repeated push-pulls and thus filling and emptying of the biosensor chamber. Furthermore, by addressing the issue of rotational speed dependency and limited space concerns in implementing repetitive filling and evacuation steps, this newly introduced technique increases the flexibility of the microfluidic CD platform to perform multi-step biological and chemical processes.

  11. Sequential push-pull pumping mechanism for washing and evacuation of an immunoassay reaction chamber on a microfluidic CD platform.

    Science.gov (United States)

    Thio, Tzer Hwai Gilbert; Ibrahim, Fatimah; Al-Faqheri, Wisam; Soin, Norhayati; Kahar Bador, Maria; Madou, Marc

    2015-01-01

    A centrifugal compact disc (CD) microfluidic platform with reservoirs, micro-channels, and valves can be employed for implementing a complete immunoassay. Detection or biosensor chambers are either coated for immuno-interaction or a biosensor chip is inserted in them. On microfluidic CDs featuring such multi-step chemical/biological processes, the biosensor chamber must be repeatedly filled with fluids such as enzymes solutions, buffers, and washing solutions. After each filling step, the biosensor chamber needs to be evacuated by a passive siphoning process to prepare it for the next step in the assay. However, rotational speed dependency and limited space on a CD are two big obstacles to performing such repetitive filling and siphoning steps. In this work, a unique thermo-pneumatic (TP) Push-Pull pumping method is employed to provide a superior alternative biosensor chamber filling and evacuation technique. The proposed technique is demonstrated on two CD designs. The first design features a simple two-step microfluidic process to demonstrate the evacuation technique, while the second design shows the filling and evacuation technique with an example sequence for an actual immunoassay. In addition, the performance of the filling and evacuation technique as a washing step is also evaluated quantitatively and compared to the conventional manual bench top washing method. The two designs and the performance evaluation demonstrate that the technique is simple to implement, reliable, easy to control, and allows for repeated push-pulls and thus filling and emptying of the biosensor chamber. Furthermore, by addressing the issue of rotational speed dependency and limited space concerns in implementing repetitive filling and evacuation steps, this newly introduced technique increases the flexibility of the microfluidic CD platform to perform multi-step biological and chemical processes.

  12. A novel fluorescence immunoassay for the sensitive detection of Escherichia coli O157:H7 in milk based on catalase-mediated fluorescence quenching of CdTe quantum dots

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Rui [College of Life Science, Nanchang University, Nanchang, 330031 (China); State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, 330047 (China); Huang, Xiaolin; Li, Juan; Shan, Shan; Lai, Weihua [State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, 330047 (China); Xiong, Yonghua, E-mail: yhxiongchen@163.com [College of Life Science, Nanchang University, Nanchang, 330031 (China); State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, 330047 (China)

    2016-12-01

    Immunoassay is a powerful tool for rapid detection of food borne pathogens in food safety monitoring. However, conventional immunoassay always suffers from low sensitivity when it employs enzyme-catalyzing chromogenic substrates to generate colored molecules as signal outputs. In the present study, we report a novel fluorescence immunoassay for the sensitive detection of E. coli O157:H7 through combination of the ultrahigh bioactivity of catalase to hydrogen peroxide (H{sub 2}O{sub 2}) and H{sub 2}O{sub 2}-sensitive mercaptopropionic acid modified CdTe QDs (MPA-QDs) as a signal transduction. Various parameters, including the concentrations of anti-E. coli O157:H7 polyclonal antibody and biotinylated monoclonal antibody, the amounts of H{sub 2}O{sub 2} and streptavidin labeled catalase (CAT), the hydrolysis temperature and time of CAT to H{sub 2}O{sub 2}, as well as the incubation time between H{sub 2}O{sub 2} and MPA-QDs, were systematically investigated and optimized. With optimal conditions, the catalase-mediated fluorescence quenching immunoassay exhibits an excellent sensitivity for E. coli O157:H7 with a detection limit of 5 × 10{sup 2} CFU/mL, which was approximately 140 times lower than that of horseradish peroxidase-based colorimetric immunoassay. The reliability of the proposed method was further evaluated using E. coli O157:H7 spiked milk samples. The average recoveries of E. coli O157:H7 concentrations from 1.18 × 10{sup 3} CFU/mL to 1.18 × 10{sup 6} CFU/mL were in the range of 65.88%–105.6%. In brief, the proposed immunoassay offers a great potential for rapid and sensitive detection of other pathogens in food quality control. - Highlights: • A novel fluorescence immunoassay was developed for the ultrasensitive detection of E. coli O157:H7. • This detection was achieved through the combination of the high bioactivity of CAT and H{sub 2}O{sub 2}-sensitive QDs. • The activity of CAT to H{sub 2}O{sub 2} is 1000 folds higher than that of the HRP

  13. Evaluation of the long-term energy analysis program used for the 1978 EIA Administrator's Report to Congress

    Energy Technology Data Exchange (ETDEWEB)

    Peelle, R. W.; Weisbin, C. R.; Alsmiller, Jr., R. G.

    1981-10-01

    An evaluation of the Long-Term Energy Analysis Program (LEAP), a computer model of the energy portion of the US economy that was used for the 1995-2020 projections in its 1978 Annual Report to Congress, is presented. An overview of the 1978 version, LEAP Model 22C, is followed by an analysis of the important results needed by its users. The model is then evaluated on the basis of: (1) the adequacy of its documentation; (2) the local experience in operating the model; (3) the adequacy of the numerical techniques used; (4) the soundness of the economic and technical foundations of the model equations; and (5) the degree to which the computer program has been verified. To show which parameters strongly influence the results and to approach the question of whether the model can project important results with sufficient accuracy to support qualitative conclusions, the numerical sensitivities of some important results to model input parameters are described. The input data are categorized and discussed, and uncertainties are given for some parameters as examples. From this background and from the relation of LEAP to other available approaches for long-term energy modeling, an overall evaluation is given of the model's suitability for use by the EIA.

  14. An Inexpensive, Fast and Sensitive Quantitative Lateral Flow Magneto-Immunoassay for Total Prostate Specific Antigen

    Science.gov (United States)

    Barnett, Jacqueline M.; Wraith, Patrick; Kiely, Janice; Persad, Raj; Hurley, Katrina; Hawkins, Peter; Luxton, Richard

    2014-01-01

    We describe the detection characteristics of a device the Resonant Coil Magnetometer (RCM) to quantify paramagnetic particles (PMPs) in immunochromatographic (lateral flow) assays. Lateral flow assays were developed using PMPs for the measurement of total prostate specific antigen (PSA) in serum samples. A detection limit of 0.8 ng/mL was achieved for total PSA using the RCM and is at clinically significant concentrations. Comparison of data obtained in a pilot study from the analysis of serum samples with commercially available immunoassays shows good agreement. The development of a quantitative magneto-immunoassay in lateral flow format for total PSA suggests the potential of the RCM to operate with many immunoassay formats. The RCM has the potential to be modified to quantify multiple analytes in this format. This research shows promise for the development of an inexpensive device capable of quantifying multiple analytes at the point-of-care using a magneto-immunoassay in lateral flow format. PMID:25587419

  15. Fluorescent Immunoassay Development for PrPSc Detection and Antemortem Diagnosis of TSEs

    National Research Council Canada - National Science Library

    Carp, Richard I

    2005-01-01

    The overall goal of our study is to develop methods of high-sensitivity and high-specificity for the antemortem diagnosis of prion diseases by detecting PrPSc in biological fluids using fluorescent immunoassay...

  16. Fluorescent Immunoassay Development for PrP(Sc) Detection and Antemortem Diagnosis of TSEs

    National Research Council Canada - National Science Library

    Carp, Richard I

    2004-01-01

    The overall goal of our study is to develop methods of high-sensitivity and high-specificity for the antemortem diagnosis of prion diseases by detecting PrPsc in biological fluids using fluorescent immunoassay...

  17. The ENZYME data bank.

    Science.gov (United States)

    Bairoch, A

    1994-09-01

    The ENZYME data bank is a repository of information relative to the nomenclature of enzymes. It is primarily based on the recommendations of the Nomenclature Committee of the International Union of Biochemistry and Molecular Biology (IUBMB) and it contains the following data for each type of characterized enzyme for which an EC (Enzyme Commission) number has been provided: EC number Recommended name Alternative names (if any) Catalytic activity Cofactors (if any) Pointers to the SWISS-PROT protein sequence entrie(s) that correspond to the enzyme (if any) Pointers to human disease(s) associated with a deficiency of the enzyme (if any).

  18. The effects of congestion of vehicles on the environment – an EIA in the Ngorongoro crater. Results from the scoping process

    OpenAIRE

    Thomassen, Jørn; Keyyu, Julius; Haaland, Hanne

    2005-01-01

    Thomassen, J., Keyyu, J., and Haaland, H. 2005. The effects of congestion of vehicles on the environment - an EIA in the Ngrongoro crates: results from the scoping process. - NINA Rapport 17. 68 s. The Ngorongoro Conservation Area Authority (NCAA) faces a great challenge in managing the vehicle congestion in the Ngorongoro crater. They are in need for an assessment of the present situation and realistic plans for mitigating measures and other management actions. The situation is also dealt wi...

  19. Control of (pre-analytical aspects in immunoassay measurements of metabolic hormones in rodents

    Directory of Open Access Journals (Sweden)

    Maximilian Bielohuby

    2018-04-01

    Full Text Available The measurement of circulating hormones by immunoassay remains a cornerstone in preclinical endocrine research. For scientists conducting and interpreting immunoassay measurements of rodent samples, the paramount aim usually is to obtain reliable and meaningful measurement data in order to draw conclusions on biological processes. However, the biological variability between samples is not the only variable affecting the readout of an immunoassay measurement and a considerable amount of unwanted or unintended variability can be quickly introduced during the pre-analytical and analytical phase. This review aims to increase the awareness for the factors ‘pre-analytical’ and ‘analytical’ variability particularly in the context of immunoassay measurement of circulating metabolic hormones in rodent samples. In addition, guidance is provided how to gain control over these variables and how to avoid common pitfalls associated with sample collection, processing, storage and measurement. Furthermore, recommendations are given on how to perform a basic validation of novel single and multiplex immunoassays for the measurement of metabolic hormones in rodents. Finally, practical examples from immunoassay measurements of plasma insulin in mice address the factors ‘sampling site and inhalation anesthesia’ as frequent sources of introducing an unwanted variability during the pre-analytical phase. The knowledge about the influence of both types of variability on the immunoassay measurement of circulating hormones as well as strategies to control these variables are crucial, on the one hand, for planning and realization of metabolic rodent studies and, on the other hand, for the generation and interpretation of meaningful immunoassay data from rodent samples.

  20. Screening for antibodies against Treponema pallidum with chemiluminescent microparticle immunoassay: analysis of discordant serology results and clinical characterization.

    Science.gov (United States)

    Li, Zhiyan; Feng, Zhenru; Liu, Ping; Yan, Cunling

    2016-09-01

    Traditionally, testing for syphilis has consisted of initial screening with a non-treponemal test, then retesting reactive specimens with a treponemal test. Recent availability of a chemiluminescent microparticle immunoassay for detecting antibodies against Treponema pallidum has led several laboratories in China to adopt chemiluminescent microparticle immunoassay for screening of syphilis, with subsequent testing of reactive serum samples with non-treponemal tests. We evaluated the utility of chemiluminescent microparticle immunoassay for routine screening of syphilis. Antibodies against Treponema pallidum were screened in 20,550 serum samples using chemiluminescent microparticle immunoassay. Chemiluminescent microparticle immunoassay-positive samples were reflexively tested with rapid plasma reagin tests and Treponema pallidum particle agglutination assays. Dot-immunoblot assays were used to confirm results of chemiluminescent microparticle immunoassay-positive and Treponema pallidum particle agglutination-negative serum samples. Overall, 267 samples (1.3%) were chemiluminescent microparticle immunoassay-positive, and 185 (69.3%) of those chemiluminescent microparticle immunoassay-positive serum samples were also Treponema pallidum particle agglutination-positive. Samples' signal to cut-off ratio for chemiluminescent microparticle immunoassay correlated with diagnostic reliability, as greater samples' signal to cut-off ratio corresponded with greater concordance between chemiluminescent microparticle immunoassay and Treponema pallidum particle agglutination results. Dot-immunoblot testing of 82 chemiluminescent microparticle immunoassay-positive and Treponema pallidum particle agglutination-negative serum samples showed that 16 samples (19.5%) were Dot-immunoblot-positive, 28 (34.2%) were indeterminate and 38 (46.3%) were negative. Because there is a certain percentage of false-positive results using chemiluminescent microparticle immunoassay for routine

  1. Determining adrenocortical activity as a measure of stress in African ...

    African Journals Online (AJOL)

    Little is known about the levels of stress experienced by African buffalos affected by injury, disease, or socio-ecological and anthropogenic factors. To be able to start filling this gap, we examined the suitability of two 11-oxoaetiocholanolone enzyme-immunoassays (EIAs) detecting 11,17 dioxoandrostanes (11,17-DOA) as ...

  2. The added value of hepatitis E diagnostics in determining causes of hepatitis in routine diagnostic settings in the Netherlands

    NARCIS (Netherlands)

    Doting, M. H. E.; Weel, J.; Niesters, H. G. M.; Riezebos-Brilman, A.; Brandenburg, A.

    OBJECTIVES: Hepatitis E virus (HEV) genotype 3 is endemic in Europe and an underdiagnosed and emerging (public) health issue. In recent years commercial enzyme immunoassays (EIAs) that detect antibodies to HEV more adequately, became available. We investigated the added value of this HEV serology in

  3. Application of porous foams for size-selective measurements of airborne wheat allergen

    NARCIS (Netherlands)

    Bogdanovic, J.; Pater, A.J. de; Doekes, G.; Wouters, I.M.; Heederik, D.J.J.

    2006-01-01

    Background: Exposure to airborne wheat allergen is a well-known cause of bakers' allergy and asthma. Airborne wheat allergen can be measured by enzyme immunoassays (EIAs) in extracts of inhalable dust samples, but only limited knowledge is available on the size distribution of wheat

  4. Comparison of neutralizing and hemagglutination-inhibiting antibody responses to influenza A virus vaccination of human immunodeficiency virus-infected individuals

    NARCIS (Netherlands)

    Benne, CA; Harmsen, M; Tavares, L; Kraaijeveld, CA; De Jong, JC

    A neutralization enzyme immunoassay (N-EIA) was used to determine the neutralizing serum antibody titers to influenza A/Taiwan/1/86 (H1N1) and Beijing/353/89 (H3N2) viruses after vaccination of 51 human immunodeficiency virus (HIV) type 1-infected individuals and 10 healthy noninfected controls

  5. Circulation of HIV antigen in blood according to stage of infection, risk group, age and geographic origin

    NARCIS (Netherlands)

    Goudsmit, J.; Paul, D. A.

    1987-01-01

    Human immunodeficiency virus antigen (HIV-ag) was determined by enzyme immunoassay (EIA) in HIV-antibody (anti-HIV) positive as well as pre-anti-HIV seroconversion sera and the results analysed according to stage of infection, risk group, age and geographic origin. Eleven (19%) of 58 homosexual men

  6. Final disposal of spent nuclear fuel in Sweden. Some unresolved issues and challenges in the design and implementation of the forthcoming planning and EIA processes

    International Nuclear Information System (INIS)

    Bjarnadottir, H.; Hilding-Rydevik, T.

    2001-06-01

    The aim of the study is to highlight some unresolved and challenging issues in the forthcoming approximately six year long Environmental Impact Assessment (EIA) and planning process of the long-term disposal of spent nuclear fuel in Sweden. Different international and Nordic experiences of the processes for final disposal as well as from other development of similar scope, where experiences assumed to be of importance for final disposal of nuclear waste, have been described. Furthermore, issues relating to 'good EIA practice' as well as certain aspects of planning theory have also been presented. The current Swedish situation for the planning and EIA process of the final disposal of spent nuclear fuel was also been summarized. These different 'knowledge areas' have been compared and measured against our perception of the expectations towards the forthcoming process, put forward by different Swedish actors in the field. The result is a presentation of a number of questions and identification issues that the authors consider need special attention in the design and conduction of the planning and EIA process. The study has been realized through a literature survey and followed by reading and analysis of the written material. The main focus of the literature search was on material describing planning processes, actor perspectives and EIA. Material and literature on the technical and scientific aspects of spent nuclear fuel disposal was however deliberately avoided. There is a wealth of international and Swedish literature concerning final disposal of spent nuclear fuel - concerning both technical issues and issues concerning for example public participation and risk perception. But material of a more systematic and comparative nature (relating to both empirical and theoretical issues, and to practical experiences) in relation to EIA processes and communicative planning for final disposal of spent nuclear fuel seems to be more sparsely represented. Our perception of

  7. Analytical evaluation of the novel Lumipulse G BRAHMS procalcitonin immunoassay

    Directory of Open Access Journals (Sweden)

    Orazio Ruzzenente

    2016-12-01

    Full Text Available Objectives: This study was designed to evaluate the analytical performance of the novel Lumipulse G1200 BRAHMS procalcitonin (PCT immunoassay. Design and methods: This analytical evaluation encompassed the calculation of the limit of blank (LOB, limit of detection (LOD, functional sensitivity, intra- and inter-assay imprecision, confirmation of linearity and a comparison with the Vidas BRAHMS PCT assay. Results: The LOB, LOD and functional sensitivity were 0.0010 ng/mL, 0.0016 ng/mL and 0.008 ng/mL, respectively. The total analytical imprecision was found to be 2.1% and the linearity was excellent (r=1.00 in the range of concentrations between 0.006–75.5 ng/mL. The correlation coefficient with Vidas BRAHMS PCT was 0.995 and the equation of the Passing and Bablok regression analysis was [Lumipulse G BRAHMS PCT]=0.76×[Vidas BRAHMS PCT]+0.04. The mean overall bias of Lumipulse G BRAHMS PCT versus Vidas BRAHMS PCT was −3.03 ng/mL (95% confidence interval [CI]: −4.32 to −1.74 ng/mL, whereas the mean bias in samples with PCT concentration between 0–10 ng/mL was −0.49 ng/mL (95% CI: −0.77 to −0.24 ng/mL. The diagnostic agreement was 100% at 0.5 ng/mL, 97% at 2.0 ng/mL and 95% at 10 ng/mL, respectively. Conclusions: These results attest that Lumipulse G BRAHMS PCT exhibits excellent analytical performance, among the best of the methods currently available on the diagnostic market. However, the significant bias compared to the Vidas BRAHMS PCT suggests that the methods cannot be used interchangeably. Keywords: Sepsis, Infection, Procalcitonin, Immunoassay

  8. Catch and measure-mass spectrometry-based immunoassays in biomarker research.

    Science.gov (United States)

    Weiß, Frederik; van den Berg, Bart H J; Planatscher, Hannes; Pynn, Christopher J; Joos, Thomas O; Poetz, Oliver

    2014-05-01

    Mass spectrometry-based (MS) methods are effective tools for discovering protein biomarker candidates that can differentiate between physiological and pathophysiological states. Promising candidates are validated in studies comprising large patient cohorts. Here, targeted protein analytics are used to increase sample throughput. Methods involving antibodies, such as sandwich immunoassays or Western blots, are commonly applied at this stage. Highly-specific and sensitive mass spectrometry-based immunoassays that have been established in recent years offer a suitable alternative to sandwich immunoassays for quantifying proteins. Mass Spectrometric ImmunoAssays (MSIA) and Stable Isotope Standards and Capture by Anti-Peptide Antibodies (SISCAPA/iMALDI) are two prominent types of MS-based immunoassays in which the capture is done either at the protein or the peptide level. We present an overview of these emerging types of immunoassays and discuss their suitability for the discovery and validation of protein biomarkers. This article is part of a Special Issue entitled: Biomarkers: A Proteomic Challenge. © 2013.

  9. Do all screening immunoassay positive buprenorphine samples need to be confirmed?

    Science.gov (United States)

    Saleem, Mohamed; Martin, Helen; Tolya, Anne; Coates, Penny

    2017-11-01

    Background Interference from opiates in the Microgenics CEDIA® Buprenorphine assay is known to produce false-positive buprenorphine screening immunoassay results necessitating confirmatory buprenorphine testing by chromatography/mass spectrometry methods. Method We reviewed data on falsely positive buprenorphine immunoassay screen (cut-off ≥ 5  µg/L) but negative for buprenorphine by gas chromatography mass spectrometry (cut-off ≥ 5  µg/L) and had a positive opiate immunoassay result (cut-off ≥ 300  µg/L). The results were collected over three months, and the data were evaluated to determine whether there is an opiate immunoassay screen concentration below which a false-positive buprenorphine result will not occur. Results We found that cross-reactivity in the CEDIA® buprenorphine immunoassay by opiates at concentrations buprenorphine result. After changing our practice to not proceed with confirmatory buprenorphine gas chromatography mass spectrometry assay when the opiate screening concentration is below an even more conservative cut-off of buprenorphine immunoassay screen do not require confirmatory testing for buprenorphine.

  10. Enzyme inhibition by iminosugars

    DEFF Research Database (Denmark)

    López, Óscar; Qing, Feng-Ling; Pedersen, Christian Marcus

    2013-01-01

    Imino- and azasugar glycosidase inhibitors display pH dependant inhibition reflecting that both the inhibitor and the enzyme active site have groups that change protonation state with pH. With the enzyme having two acidic groups and the inhibitor one basic group, enzyme-inhibitor complexes...

  11. Basic and clinical investigation of T3 immunoassay kit

    International Nuclear Information System (INIS)

    Konishi, Junji; Nakajima, Akiko; Morita, Rikushi; Endo, Keigo; Ikekubo, Katsuji

    1976-01-01

    T 3 immunoassay kit was investigated basically and clinically. A good result was obtained at the prescribed incubation temperature and for 16 hours of incubation time. Moreover, it was thought to be possible that incubation time could be shortened to 1 - 4 hours at 37 0 C. Specificity of antibody was good. Recovery of added T 3 was 100+-5 (S.D.) % on an average and parallel of dilution curve of high T 3 serum was also good. Variation coefficient of accuracy of this kit was 1.5 - 2.1 % and that of reproducibility was 1.3 - 6.6 %. Mild hemolysis did not affect measurement value. Serum T 3 level in normals, untreated patients with Basedow's disease and patients with primary hypothyroidism was 142+-21 ng/100 ml, 452+-156 ng/100 ml and 67+-17 ng/100 ml, respectively. Serum T 3 level in patients with Hashimoto's disease was distributed to a wide extent, but that of patients with goiter and simple goiter ranged within normal range. On the other side, serum T 3 level of normal pregnant woman was high and that of patients with anorexia nervosa showed low level. From the above mentioned results, it was concluded that this kit was simple in method and good in sensitivity, specificity and reproducibility and it was also useful for clinical applications. (M. Tsunoda)

  12. Backscattering particle immunoassays in wire-guide droplet manipulations

    Directory of Open Access Journals (Sweden)

    You David J

    2008-11-01

    Full Text Available Abstract A simpler way for manipulating droplets on a flat surface was demonstrated, eliminating the complications in the existing methods of open-surface digital microfluidics. Programmed and motorized movements of 10 μL droplets were demonstrated using stepper motors and microcontrollers, including merging, complicated movement along the programmed path, and rapid mixing. Latex immunoagglutination assays for mouse immunoglobulin G, bovine viral diarrhea virus and Escherichia coli were demonstrated by merging two droplets on a superhydrophobic surface (contact angle = 155 ± 2° and using subsequent back light scattering detection, with detection limits of 50 pg mL-1, 2.5 TCID50 mL-1 and 85 CFU mL-1, respectively, all significantly lower than the other immunoassay demonstrations in conventional microfluidics (~1 ng mL-1 for proteins, ~100 TCID50 mL-1 for viruses and ~100 CFU mL-1 for bacteria. Advantages of this system over conventional microfluidics or microwell plate assays include: (1 minimized biofouling and repeated use (>100 times of a platform; (2 possibility of nanoliter droplet manipulation; (3 reprogrammability with a computer or a game pad interface.

  13. Designing novel nano-immunoassays: antibody orientation versus sensitivity

    Energy Technology Data Exchange (ETDEWEB)

    Puertas, S; Moros, M; Fernandez-Pacheco, R; Ibarra, M R; Grazu, V; De la Fuente, J M, E-mail: vgrazu@unizar.e, E-mail: jmfuente@unizar.e [Instituto de Nanociencia de Aragon (INA), Universidad de Zaragoza, Campus RIo Ebro, EdifIcio I-D, Mariano Esquillor, s/n, 50018 Zaragoza (Spain)

    2010-12-01

    There is a growing interest in the use of magnetic nanoparticles (MNPs) for their application in quantitative and highly sensitive biosensors. Their use as labels of biological recognition events and their detection by means of some magnetic method constitute a very promising strategy for quantitative high-sensitive lateral-flow assays. In this paper, we report the importance of nanoparticle functionalization for the improvement of sensitivity for a lateral-flow immunoassay. More precisely, we have found that immobilization of IgG anti-hCG through its polysaccharide moieties on MNPs allows more successful recognition of the hCG hormone. Although we have used the detection of hCG as a model in this work, the strategy of binding antibodies to MNPs through its sugar chains reported here is applicable to other antibodies. It has huge potential as it will be very useful for the development of quantitative and high-sensitive lateral-flow assays for its use on human and veterinary, medicine, food and beverage manufacturing, pharmaceutical, medical biologics and personal care product production, environmental remediation, etc.

  14. A Multiplex Microsphere Immunoassay for Zika Virus Diagnosis

    Directory of Open Access Journals (Sweden)

    Susan J. Wong

    2017-02-01

    Full Text Available Rapid and accurate diagnosis of infectious agents is essential for patient care, disease control, and countermeasure development. The present serologic diagnosis of Zika virus (ZIKV infection relies mainly on IgM-capture ELISA which is confounded with the flaw of cross-reactivity among different flaviviruses. In this communication, we report a multiplex microsphere immunoassay (MIA that captures the diagnostic power of viral envelope protein (that elicits robust, yet cross-reactive antibodies to other flaviviruses and the differential power of viral nonstructural proteins NS1 and NS5 (that induce more virus-type specific antibodies. Using 153 patient specimens with known ZIKV and/or dengue virus (DENV; a closely related flavivirus infections, we showed that (i ZIKV envelope-based MIA is equivalent or more sensitive than IgM-capture ELISA in diagnosing ZIKV infection, (ii antibody responses to NS1 and NS5 proteins are more ZIKV-specific than antibody response to envelope protein, (iii inclusion of NS1 and NS5 in the MIA improves the diagnostic accuracy when compared with the MIA that uses envelope protein alone. The multiplex MIA achieves a rapid diagnosis (turnaround time < 4 h and requires small specimen volume (10 μl in a single reaction. This serologic assay could be developed for use in clinical diagnosis of ZIKV infection and for monitoring immune responses in vaccine trials.

  15. A Multiplex Microsphere Immunoassay for Zika Virus Diagnosis.

    Science.gov (United States)

    Wong, Susan J; Furuya, Andrea; Zou, Jing; Xie, Xuping; Dupuis, Alan P; Kramer, Laura D; Shi, Pei-Yong

    2017-02-01

    Rapid and accurate diagnosis of infectious agents is essential for patient care, disease control, and countermeasure development. The present serologic diagnosis of Zika virus (ZIKV) infection relies mainly on IgM-capture ELISA which is confounded with the flaw of cross-reactivity among different flaviviruses. In this communication, we report a multiplex microsphere immunoassay (MIA) that captures the diagnostic power of viral envelope protein (that elicits robust, yet cross-reactive antibodies to other flaviviruses) and the differential power of viral nonstructural proteins NS1 and NS5 (that induce more virus-type specific antibodies). Using 153 patient specimens with known ZIKV and/or dengue virus (DENV; a closely related flavivirus) infections, we showed that (i) ZIKV envelope-based MIA is equivalent or more sensitive than IgM-capture ELISA in diagnosing ZIKV infection, (ii) antibody responses to NS1 and NS5 proteins are more ZIKV-specific than antibody response to envelope protein, (iii) inclusion of NS1 and NS5 in the MIA improves the diagnostic accuracy when compared with the MIA that uses envelope protein alone. The multiplex MIA achieves a rapid diagnosis (turnaround timeimmune responses in vaccine trials. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

  16. Chronoamperometry-Based Redox Cycling for Application to Immunoassays.

    Science.gov (United States)

    Lee, Ga-Yeon; Park, Jun-Hee; Chang, Young Wook; Cho, Sungbo; Kang, Min-Jung; Pyun, Jae-Chul

    2018-01-26

    In this work, the chronoamperometry-based redox cycling of 3,3',5,5'-tetramethylbenzidine (TMB) was performed by using interdigitated electrode (IDE). The signal was obtained from two sequential chronoamperometric profiles: (1) with the generator at the oxidative potential of TMB and the collector at the reductive potential of TMB, and (2) with the generator at the reductive potential of TMB and the collector at the oxidative potential of TMB. The chronoamperometry-based redox cycling (dual mode) showed a sensitivity of 1.49 μA/OD, and the redox cycling efficiency was estimated to be 94% (n = 10). The sensitivities of conventional redox cycling with the same interdigitated electrode and chronoamperometry using a single working electrode (single mode) were estimated to be 0.67 μA/OD and 0.18 μA/OD, respectively. These results showed that the chronoamperometry-based redox cycling (dual mode) could be more effectively used to quantify the oxidized TMB than other amperometric methods. The chronoamperometry-based redox cycling (dual mode) was applied to immunoassays using a commercial ELISA kit for medical diagnosis of the human hepatitis B virus surface antigen (hHBsAg). Finally, the chronoamperometry-based redox cycling (dual mode) provided more than a 10-fold higher sensitivity than conventional chronoamperometry using a single working electrode (single mode) when applied to a commercial ELISA kit for medical diagnosis of hHBsAg.

  17. Evaluation of a new chemiluminescence immunoassay for diagnosis of syphilis

    Directory of Open Access Journals (Sweden)

    Mo Xiaohui

    2010-02-01

    Full Text Available Abstract Objective To assess the sensitivity, specificity, and feasibility of a new chemiluminescence immunoassay (CLIA in the diagnosis of syphilis. Methods At first, a retrospective study was conducted, using 135 documented cases of syphilis and 30 potentially interfering samples and 80 normal sera. A prospective study was also performed by testing 2, 071 unselected samples for routine screening for syphilis. CLIA was compared with a nontreponemal test (TRUST and a treponemal test (TPPA. Results There was an agreement of 100% between CLIA and TPPA in the respective study. The percentage of agreement among the 245 sera tested was 100.0%. Compared with TPPA, the specificity of CLIA was 99.9% (1817/1819, the sensitivity of CLIA was 100.0% (244/244 in the prospective study. CLIA showed 99.5% agreement with TPPA by testing 2, 071 unselected samples. And CLIA seemed to be more sensitive than TPPA in detecting the samples of primary syphilis. Conclusions CLIA is easy to perform and the indicator results are objective and unequivocal. It may be suitable for large-scale screening as a treponemal test substituted for TPPA.

  18. A rapid lateral flow immunoassay for serological diagnosis of pertussis.

    Science.gov (United States)

    Salminen, Teppo; Knuutila, Aapo; Barkoff, Alex-Mikael; Mertsola, Jussi; He, Qiushui

    2018-03-07

    Current serological diagnosis of pertussis is usually done by ELISA. However, the ELISAs are often central-laboratory based, require trained staff and have long turnaround times. A rapid point-of-care (POC) assay for pertussis serology would aid in both diagnosis and surveillance of the disease. While lateral flow immunoassays (LFIA) are simple to use and ideal for point-of-care diagnostics, they were limited to qualitative assays until recently. In this study, we developed a quantitative LFIA with fluorescent Eu-nanoparticle reporters for the detection of anti-pertussis toxin (PT) IgG. The assay was evaluated by testing 198 serum samples with varying anti-PT IgG levels and the result was compared to those obtained with standardized anti-PT IgG ELISA. At the diagnostic cutoff of 100 IU/mL in ELISA, the LFIA had a concordance of 92% with the ELISA, with a specificity of 96% [95% confidence interval (CI): 89-99%] and a sensitivity of 88% [CI: 77-94%]. The developed LFIA has a turnaround time of one hour and requires only a simple manipulation by the user and an instrument for the quantitative detection of the signal. We conclude that the LFIA is specific and sensitive for serological diagnosis of pertussis and is suitable for a POC test. Copyright © 2018 Elsevier Ltd. All rights reserved.

  19. PhoneQuant: A smartphone-based quantitative immunoassay analyser.

    Science.gov (United States)

    Shah, Malay Ilesh; Joseph, Jayaraj; Sanne, Ujwal Sriharsha; Sivaprakasam, Mohanasankar

    2017-07-01

    There is a vital need for portable and cost-effective point-of-care (PoC) testing technologies that provide reliable and rapid results. Lateral Flow Immunoassays (LFIA) are suitable PoC diagnostic tools with the potential for use in a wide variety of field applications ranging from uses in clinical diagnostics to aiding law enforcement. Quick and reliable diagnosis of non-communicable diseases (NCD) like diabetes is vital especially in developing countries like India where the burden of these diseases is very high and is increasing day by day. In this paper, we have presented the design of smartphone-based fully quantitative LFIA analyser, An automatic image processing algorithm is also described. A repeatability study was done with stable fluorescence reference cartridges. The Coefficient of Variation (CoV) for repeatability study was calculated and it was found to be good (LFIA analysers and it has good potential to be deployed at physician's desk or for in-home PoC testing for quick and reliable diagnosis.

  20. Using Compliance Analysis for PPP to bridge the gap between SEA and EIA: Lessons from the Turcot Interchange reconstruction in Montréal, Québec

    International Nuclear Information System (INIS)

    Thompson, Undiné-Celeste; Marsan, Jean-François; Fournier-Peyresblanques, Bastien; Forgues, Chantal; Ogaa, Anita; Jaeger, Jochen A.G.

    2013-01-01

    There is increasing concern about the disjunct between the intent of higher level government goals and actual projects “on the ground” in Canada. Although strategic environmental assessment (SEA) and a wide variety of plans, policies and programmes (PPP) contain and promote goals that envision a movement towards social, economic and environmental sustainability, these goals are not necessarily upheld by large-scale projects and their environmental impact assessments (EIAs). This disconnect is often illustrated through anecdotal observations. However, to be able to overcome this disjunct it is imperative to come to a clearer understanding of the degree of sustainability or unsustainability of large-scale developments and the way in which they “measure up” in terms of the goals when compared to alternative options. This article proposes a Compliance Analysis method for investigating the level of harmonization between SEA, PPP and proposed projects and their possible alternatives (CAPPP). This method is quantified through a Likert scale which allows for comparison of alternatives for decision making and analytical purposes. The 2009 proposal for the Turcot Exchange redevelopment in Montréal, Québec, put forward by the Ministry of Transport of Québec (MTQ), as well as two alternative proposals, were utilized as a case study to clearly demonstrate the CAPPP methodology and its applicability. The approved plan for the Turcot redevelopment proposed by MTQ was found to be in poor compliance with the majority of the 178 goals in the six sectors that were examined (air quality, climate change, health, noise, socioeconomic, transport), while alternative proposals were found to be in greater accordance with the intentions of governmental SEA and PPP. Synthesis and applications: The CAPPP methodology is a versatile “watchdog” tool for the examination of the level of compliance between stated goals for regions, industrial sectors, or governments and the EIAs of