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Validation of a blocking enzyme-linked immunosorbent assay for detection of antibodies against porcine reproductive and respiratory syndrome virus.
- Source :
-
Clinical and diagnostic laboratory immunology [Clin Diagn Lab Immunol] 2004 May; Vol. 11 (3), pp. 503-14. - Publication Year :
- 2004
-
Abstract
- Porcine reproductive and respiratory syndrome (PRRS) continues to be one of the most significant diseases of swine. IDEXX HerdChek PRRS, a commercially available enzyme-linked immunosorbent assay (ELISA), has become the industry standard for the detection of antibodies against PRRS virus (PRRSV). The need to accurately determine the PRRSV serostatus of herds and individual animals has prompted the development of several follow-up assay methods. A highly specific and repeatable blocking ELISA (bELISA) was developed on the basis of the use of an expressed PRRSV nucleocapsid (N) protein as the antigen and a biotinylated monoclonal antibody. Validation of the bELISA used sera from 316 animals experimentally and naturally infected with North American PRRSV and sera from 370 uninfected animals. Receiver operating characteristic analysis of the data calculated a diagnostic sensitivity of 97.8% and a diagnostic specificity of 100%. The between-run coefficient of variation of an internal quality control serum was 4.24%. The bELISA was able to detect seroconversion as well as the IDEXX ELISA and the indirect fluorescent antibody (IFA) assay; kappa values were 0.94 and 0.96, respectively. A collection of 133 serum samples with unexpected positive IDEXX ELISA results was obtained from 4,038 diagnostic samples submitted from farms from which PRRS-negative results were expected. The bELISA identified 97% of the samples as PRRS seronegative, while the IFA identified 100% as seronegative. The anticipated use of the bELISA is as a follow-up test to the IDEXX ELISA for determining the PRRSV serostatus of individual animals with unexpected positive test results from swine herds from which negative results are expected.
- Subjects :
- Analysis of Variance
Animals
Antibodies, Monoclonal immunology
Antibodies, Viral immunology
Antibody Specificity immunology
Biotinylation
Blotting, Western
Confidence Intervals
Electrophoresis, Polyacrylamide Gel
Enzyme-Linked Immunosorbent Assay methods
Enzyme-Linked Immunosorbent Assay statistics & numerical data
Enzyme-Linked Immunosorbent Assay veterinary
False Positive Reactions
Infections immunology
Nucleocapsid Proteins genetics
Nucleocapsid Proteins immunology
Predictive Value of Tests
ROC Curve
Recombinant Proteins chemistry
Recombinant Proteins immunology
Reproducibility of Results
Sensitivity and Specificity
Swine
Vaccination
Antibodies, Viral blood
Porcine respiratory and reproductive syndrome virus immunology
Subjects
Details
- Language :
- English
- ISSN :
- 1071-412X
- Volume :
- 11
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- Clinical and diagnostic laboratory immunology
- Publication Type :
- Academic Journal
- Accession number :
- 15138175
- Full Text :
- https://doi.org/10.1128/CDLI.11.3.503-514.2004