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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
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Validation Study
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 Korea1College 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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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.

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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
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