Mechanization of the enzyme-linked immunosorbent assay (ELISA) for large scale screening of sera

1977 ◽  
Vol 16 (4) ◽  
pp. 351-359 ◽  
Author(s):  
E.J. Ruitenberg ◽  
J.A. van Amstel ◽  
B.J.M. Brosi ◽  
P.A. Steerenberg
Keyword(s):  
Immunosorbent Assay ◽  
Large Scale ◽  
Enzyme Linked Immunosorbent Assay ◽  
Large Scale Screening

National survey of viruses in pastures of subterranean clover. III. Equipment for large scale screening of viruses by ELISA

1993 ◽  
Vol 44 (8) ◽  
pp. 1883 ◽  
Author(s):  
LB Hulse ◽  
K Helms ◽  
PM Waterhouse
Keyword(s):  
Immunosorbent Assay ◽  
Large Scale ◽  
New South ◽  
South Australia ◽  
Subterranean Clover ◽  
Trifolium Subterraneum ◽  
Nationwide Survey ◽  
Enzyme Linked Immunosorbent Assay ◽  
South Wales ◽  
Large Scale Screening

Equipment for large scale enzyme-linked immunosorbent assay (ELISA) was developed for a 3 year nationwide survey of the incidence of four viruses in pastures of Trifolium subterraneum (subterranean clover) in New South Wales, Victoria, South Australia, Western Australia and Tasmania. Instruments were designed to (a) automate quantitatively the addition of buffer to plant samples passing through a sap extractor, (b) standardize the washing of plates in the various stages of the ELISA protocol, and (c) replace hand pipetting with a pneumatic foot-operated pipette.

Evaluation of immunoglobulin G4 subclass antibody in a peptide-based enzyme-linked immunosorbent assay for the serodiagnosis of human fascioliasis

Parasitology ◽  
2007 ◽  
Vol 134 (14) ◽  
pp. 2021-2026 ◽  
Author(s):  
C. TANTRAWATPAN ◽  
W. MALEEWONG ◽  
C. WONGKHAM ◽  
S. WONGKHAM ◽  
P. M. INTAPAN ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Large Scale ◽  
Laboratory Diagnosis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Parasitic Infections ◽  
Immunoglobulin G4 ◽  
Predictive Values ◽  
Diagnostic Potential ◽  
Human Fascioliasis ◽  
Specific Igg4

SUMMARYTo improve the diagnosis of human fascioliasis caused byFasciola gigantica, we developed a peptide-based enzyme-linked immunosorbent assay (peptide-based ELISA) based on the detection of specific IgG4 subclass antibody. Two identified B-cell epitopes ofF. giganticacathepsin L1 were synthesized as single synthetic peptides, acetyl-DKIDWRESGYVTELKDQGNC-carboxamide (peptide L) and acetyl-DKIDWRESGYVTEVKDQGNC-carboxamide (peptide V), and their diagnostic potential was evaluated. The sera of 25 patients infected withF. gigantica, 212 patients with other parasitic infections, 32 cholangiocarcinoma patients and 57 healthy controls were analysed. The sensitivity, specificity, accuracy, and positive and negative predictive values of this assay were the same with both peptides at 100%, 99·7%, 99·7%, 96·2% and 100%, respectively. These highly sensitive and specific peptide-based ELISAs for the detection of specific IgG4 antibody could be useful for laboratory diagnosis of human fascioliasis in future large-scale surveys throughout Southeast Asia where this disease is prevalent.

scholarly journals Detection of Type A, B, E, and F Clostridium botulinum Neurotoxins in Foods by Using an Amplified Enzyme-Linked Immunosorbent Assay with Digoxigenin-Labeled Antibodies

2006 ◽  
Vol 72 (2) ◽  
pp. 1231-1238 ◽  
Author(s):  
Shashi K. Sharma ◽  
Joseph. L. Ferreira ◽  
Brian S. Eblen ◽  
Richard C. Whiting
Keyword(s):  
Immunosorbent Assay ◽  
Clostridium Botulinum ◽  
Botulinum Neurotoxin ◽  
Large Scale ◽  
Lethal Dose ◽  
Meat Products ◽  
Type A ◽  
Detection Sensitivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Botulinum Neurotoxins

ABSTRACT An amplified enzyme-linked immunosorbent assay (ELISA) for the detection of Clostridium botulinum complex neurotoxins was evaluated for its ability to detect these toxins in food. The assay was found to be suitable for detecting type A, B, E, and F botulinum neurotoxins in a variety of food matrices representing liquids, solid, and semisolid food. Specific foods included broccoli, orange juice, bottled water, cola soft drinks, vanilla extract, oregano, potato salad, apple juice, meat products, and dairy foods. The detection sensitivity of the test for these botulinum complex serotypes was found to be 60 pg/ml (1.9 50% lethal dose [LD50]) for botulinum neurotoxin type A (BoNT/A), 176 pg/ml (1.58 LD50) for BoNT/B, 163 pg/ml for BoNT/E (4.5 LD50), and 117 pg/ml for BoNT/F (less than 1 LD50) in casein buffer. The test could also readily detect 2 ng/ml of neurotoxins type A, B, E, and F in a variety of food samples. For specificity studies, the assay was also used to test a large panel of type A C. botulinum, a smaller panel of proteolytic and nonproteolytic type B, E, and F neurotoxin-producing Clostridia, and nontoxigenic organisms using an overnight incubation of toxin production medium. The assay appears to be an effective tool for large-scale screening of the food supply in the event of a botulinum neurotoxin contamination event.

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

2005 ◽  
Vol 59 (3-4) ◽  
pp. 363-370
Author(s):  
Tadej Malovrh ◽  
M. Pate ◽  
M. Ocepek ◽  
B. Krt
Keyword(s):  
Antibody Response ◽  
Immunosorbent Assay ◽  
Large Scale ◽  
Serological Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Agar Gel ◽  
Bovine Leukaemia Virus ◽  
Leukaemia Virus ◽  
Elisa Kit ◽  
Pcr Method

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

scholarly journals Large-Scale Survey of CampylobacterSpecies in Human Gastroenteritis by PCR and PCR–Enzyme-Linked Immunosorbent Assay

1999 ◽  
Vol 37 (12) ◽  
pp. 3860-3864 ◽  
Author(s):  
A. J. Lawson ◽  
J. M. J. Logan ◽  
G. L. O'neill ◽  
M. Desai ◽  
J. Stanley
Keyword(s):  
Immunosorbent Assay ◽  
Large Scale ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mixed Infections ◽  
Identification Algorithm ◽  
Detection And Identification ◽  
Significant Difference ◽  
Sporadic Cases ◽  
Low Incidence ◽  
Campylobacter Infection

A PCR-based study of the incidence of enteropathogenic campylobacter infection in humans was done on the basis of a detection and identification algorithm consisting of screening PCRs and species identification by PCR-enzyme-linked immunosorbent assay. This was applied to DNA extracted from 3,738 fecal samples from patients with sporadic cases of acute gastroenteritis, submitted by seven regional Public Health Laboratories in England and Wales over a 2-year period. The sending laboratories had cultured “Campylobacterspp.” from 464 samples. The PCR methodologies detected 492Campylobacter-positive samples, and the combination of culture and PCR yielded 543 Campylobacter-positive samples. There was identity (overlap) for 413 samples, but 79 PCR-positive samples were culture negative, and 51 culture-positive samples were PCR negative. While there was no statistically significant difference between PCR and culture in detection of C. jejuni-C. coli(PCR, 478 samples; culture, 461 samples), PCR provided unique data about mixed infections and non-C. jejuni and non- C. coli campylobacters. Mixed infections withC. jejuni and C. coli were found in 19 samples, and mixed infection with C. jejuni and C. upsaliensis was found in one sample; this was not apparent from culture. Eleven cases of gastroenteritis were attributed to C. upsaliensis by PCR, three cases were attributed to C. hyointestinalis, and one case was attributed to C. lari. This represents the highest incidence of C. hyointestinalis yet reported from human gastroenteritis, while the low incidence of C. larisuggests that it is less important in this context.

scholarly journals Multiserotype Enzyme-Linked Immunosorbent Assay as a Diagnostic Aid for Periodontitis in Large-Scale Studies

2002 ◽  
Vol 40 (2) ◽  
pp. 512-518 ◽  
Author(s):  
P. J. Pussinen ◽  
T. Vilkuna-Rautiainen ◽  
G. Alfthan ◽  
K. Mattila ◽  
S. Asikainen
Keyword(s):  
Immunosorbent Assay ◽  
Large Scale ◽  
Enzyme Linked Immunosorbent Assay ◽  
Diagnostic Aid

scholarly journals Development and Application of a Competitive Enzyme-Linked Immunosorbent Assay for the Detection of Serum Antibodies to Porcine Circovirus Type 2

2000 ◽  
Vol 12 (5) ◽  
pp. 400-405 ◽  
Author(s):  
Ian W. Walker ◽  
Carrie A. Konoby ◽  
Victoria A. Jewhurst ◽  
Irene McNair ◽  
Francis McNeilly ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Large Scale ◽  
Porcine Circovirus Type ◽  
Postweaning Multisystemic Wasting Syndrome ◽  
Porcine Circovirus Type 2 ◽  
Porcine Circovirus ◽  
Enzyme Linked Immunosorbent Assay ◽  
The Mean ◽  
Immunofluorescent Test

We report the development of a competitive enzyme-linked immunosorbent assay (c-ELISA) for the detection of antibodies to porcine circovirus type 2 (PCV2), the agent associated with the recently described postweaning multisystemic wasting syndrome in pigs. At present, no method has been published describing a c-ELISA for the detection of antibodies to PCV2, and currently employed tests are impractical for use in some laboratories. The assay described here uses a cell culture isolate of porcine circovirus type 2 as antigen and a PCV2-specific monoclonal antibody as the competing reagent. Evaluation of the ELISA was performed by comparison with results obtained using an indirect immunofluorescent test on 484 sera from pig herds in the United Kingdom, Canada, France, and the USA and serial bleeds from pigs experimentally infected with porcine circoviruses. The sensitivity and specificity of the ELISA were determined as 99.58% and 97.14%, respectively, at 2 standard deviations (SD) from the mean or 95.81% and 100% at 3 SD from the mean. Using this ELISA, a serologic survey of 461 sera collected from commercial pig herds in Northern Ireland between 1973 and 1999 was undertaken. Analysis of the results of this survey demonstrated that the number of ELISA-positive sera detected in an individual year during this period ranged from 55% to 100%. This c-ELISA has applications for large-scale rapid diagnosis of PCV2 infection in pig populations worldwide and for immunoscreening of sera from other species for antibodies to PCV2.

Sensitive Measurement of Thrombopoietin by a Monoclonal Antibody Based Sandwich Enzyme-linked Immunosorbent Assay

1997 ◽  
Vol 78 (04) ◽  
pp. 1262-1267 ◽  
Author(s):  
Claudia C Folman ◽  
Albert E G K von dem Borne ◽  
Irma H J A M Rensink ◽  
Winald Gerritsen ◽  
C Ellen van der Schoot ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Immunosorbent Assay ◽  
Platelet Transfusion ◽  
Large Scale ◽  
Limit Of Detection ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Reliable Tool

SummaryIn this report a sensitive enzyme-linked immunosorbent assay (ELISA) for the measurement of plasma thrombopoietin (Tpo) is described that is solely based on monoclonal antibodies (MoAbs).The assay has an intra and inter-assay variance of 5-7% and 7-13%, respectively. Native and recombinant human Tpo (rhTpo) were recognized equally well, no cross reactivity with other cytokines was found and rhTpo added to plasma and serum was completely recovered. With the ELISA, Tpo concentrations in EDTA-anticoagulated plasma of all controls (n = 193) could be determined, since the limit of detection (2 ± 0.8 A.U./ml, mean ± sd) was lower than the concentration found in controls (11 ± 8 A.U./ml, mean ± sd; 2.5th-97.5th percentile: 4-32 A.U./ml). Tpo levels in serum were on average 3.4 times higher than in plasma.We showed in vivo that Tpo is bound by platelets, as in thrombocytopenic patients (n = 5) a platelet transfusion immediately led to a drop in plasma Tpo level, whereas in patients receiving chemotherapy the induced thrombocytopenia was followed by a rise in plasma Tpo levels.In summary, these results indicate that this ELISA is a reliable tool for Tpo measurements and is applicable for large scale studies.

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