Development and Validation of a Recombinant Nucleocapsid Protein-Based ELISA for Detection of the Antibody to Porcine Reproductive and Respiratory Syndrome Virus

Jia-Qi Chu; Xu-Min Hu; Myung-Cheol Kim; Chang-Sik Park; Moo-Hyung Jun

doi:10.1007/s12275-009-0033-x

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Development and Validation of a Recombinant Nucleocapsid Protein-Based ELISA for Detection of the Antibody to Porcine Reproductive and Respiratory Syndrome Virus: Jia-Qi Chu ¹, Xu-Min Hu ¹, Myung-Cheol Kim ¹, Chang-Sik Park ², Moo-Hyung Jun ¹; Journal of Microbiology 2009;47(5):582-588.
DOI: https://doi.org/10.1007/s12275-009-0033-x
Published online: October 24, 2009

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

Corresponding author: Moo-Hyung Jun , Tel: 82-42-821-6753,

Received: 5 February 2009 • Accepted: 20 May 2009

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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.

Keywords: PRRSV;nucleocapsid protein;indirect ELISA

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Development and Validation of a Recombinant Nucleocapsid Protein-Based ELISA for Detection of the Antibody to Porcine Reproductive and Respiratory Syndrome Virus

J. Microbiol. 2009;47(5):582-588. Published online October 24, 2009

DOI: https://doi.org/10.1007/s12275-009-0033-x

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