Development and validation of a recombinant nucleocapsid protein-based ELISA for detection of the antibody to porcine reproductive and respiratory syndrome virus.
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.
College of Veterinary Medicine, Chungnam National University, Daejeon, Republic of Korea.
Jia-Qi
JQ
Chu
Xu-Min
XM
Hu
Myung-Cheol
MC
Kim
Chang-Sik
CS
Park
Moo-Hyung
MH
Jun
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D008969:Molecular Sequence Data; D019590:Nucleocapsid Proteins / Q000276:immunology*; D019318:Porcine Reproductive and Respiratory Syndrome / Q000175:diagnosis* / Q000821:virology; D019316:Porcine respiratory and reproductive syndrome virus / Q000276:immunology* / Q000302:isolation & purification; D011237:Predictive Value of Tests; D012680:Sensitivity and Specificity; D013552:Swine
D000914:Antibodies, Viral; D019590:Nucleocapsid Proteins