PRRSV全病毒包被抗原间接ELISA抗体检测方法的建立与应用OA

Porcine reproductive and respiratory syndrome virus(PRRSV)infection seriously has endangered the sustainable and healthy development of the pigs industry in China.The anti-PRRSV antibody detection is one of the key means for disease prevention and control.To improve the accuracy of PRRSV antibody detection methods,the whole virus protein antigen of PRRSV and Marc-145 cell antigen were puri-fied by molecular sieve techniques and ultracentrifugation in this study.The SDS-PAGE and Western blot results showed that the purified PRRSV antigens mainly contained M protein and N protein,while the cell-simulated antigens did not react with the anti-PRRSV antibody.By optimizing reaction conditions,an indirect ELISA method for detecting PRRSV antibodies was established.The critical S/P value for de-termining positive and negative antibodies was 0.4,with a sensitivity of 96.47%and a specificity of 96.36%.This method showed no cross-reactivity with porcine epidemic diarrhea virus(PEDV),porcine deltacoronavirus(PDCoV),porcine circovirus type 2(PCV2),and por-cine pseudorabies virus(PRV).The repeatability tests showed that both intra-assay and inter-assay coefficients of variation were below 10%.Based on the detection results of 788 clinical serum samples,the concordance tests demonstrated that the Kappa value was 0.91,com-pared with IFA;and 0.72,compared with the commercial kit(IDEXX).In conclusion,this method is highly sensitive and specific,with high accuracy and reliability in practical applications.It would be a potential application for anti-PRRSV antibody detection and epidemiolog-ical investigation,and disease eradication.