畜牧与兽医2026，Vol.58Issue(4)：115-121,7.
PRRSV全病毒包被抗原间接ELISA抗体检测方法的建立与应用
Development and application of an indirect ELISA method for the detection of antibodies against PRRSV using whole virus coated antigens
摘要
Abstract
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.关键词
猪繁殖与呼吸综合征病毒/ELISA/全病毒抗原Key words
PRRSV/ELISA/whole virus protein antigen分类
农业科技引用本文复制引用
孙海凤,孙杨杨,白娟,姜平..PRRSV全病毒包被抗原间接ELISA抗体检测方法的建立与应用[J].畜牧与兽医,2026,58(4):115-121,7.基金项目
江苏省农业科技自主创新资金项目(CX(22)1011) （CX(22）
国家现代农业产业技术体系专项(CARS-35) （CARS-35）
