Title |
Development and Validation of a Recombinant Nucleocapsid Protein-Based ELISA for Detection of the Antibody to Porcine Reproductive and Respiratory Syndrome Virus |
Author |
Jia-Qi Chu1, Xu-Min Hu1, Myung-Cheol Kim1, Chang-Sik Park2, and Moo-Hyung Jun1* |
Address |
1College of Veterinary Medicine, Chungnam National University, Daejeon 305-764, Republic of Korea, 2Research Center for Transgenic Cloned Pigs, Chungnam National University, Daejeon 305-764, Republic of Korea |
Bibliography |
Journal of Microbiology, 47(5),582-588, 2009,
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DOI |
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Key Words |
PRRSV, nucleocapsid protein, indirect ELISA |
Abstract |
Three indirect enzyme-linked immunosorbent assays (iELISA) based on the North American like (NA-like), European like (EU-like) and co-expressed NA- and EU-like recombinant nucleocapsid proteins (N-protein) of porcine reproductive and respiratory syndrome virus (PRRSV) were validated for the detection of the antibodies in porcine sera. A total of 422 serum samples from unvaccinated pigs were tested. The cut-off value was optimized by a two-graph receiver operating characteristics analysis at a 95% confidence level. This assay was validated with Western blot analysis and IDEXX HerdChek™ ELISA. Cross-reactivity results showed that iELISA was PRRSV-specific. Repeatability tests revealed that the coefficients of variation of positive sera within and between runs were less than 10%. The results indicate that iELISA is simpler to produce and perform, time-saving and suitable for large scale surveys of PRRSV infection at low cost, and is potentially useful to evaluate the efficiency of various vaccines against PRRSV. |