PEDV流行株重组截短N蛋白间接ELISA检测方法的建立

Establishment of Indirect ELISA Method for Detecting Recombinant Truncated N Protein of Porcine Epidemic Diarrhea Virus

为了建立猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)抗体检测的间接ELISA方法,本研究以纯化的原核表达的PEDV截短N蛋白作为包被抗原,建立了PEDV抗体检测的间接ELISA方法,将该方法命名为rnPED-ELISA。该抗原不与其他常见的7种猪病的阳性血清发生交叉反应,批内和批间重复性试验的变异系数均小于13%;rnPED-ELISA相对于血清中和试验(SN)试验的敏感性为93.33%,特异性为90.00%;rnPED-ELISA与TSZ全病毒抗体检测试剂盒的符合率达91.67%。采用rnPED-ELISA方法检测200份临床样品,PEDV抗体阳性检出率为69.5%。本试验建立的rnPED-ELISA方法具有良好的敏感性和特异性,可为免疫猪群抗体监测和猪流行性腹泻流行病学调查提供一种快速、简便的血清学诊断方法。

In order to establish an indirect ELISA to detect antibody of porcine epidemic diarrhea virus（PEDV）.The experiment using the recombinant and purified truncated N protein as antigen expressed in E.coli BL21（DE3）,the indirect ELISA was named rnPED-ELISA.The recombinant truncated N protein antigen showed no cross-reaction with the positive sera of other 7kinds of swine diseases,CV%of intro-batch duplicativity test and inter-batch duplicativity test were less13%;Sensitivity and specificity of rnPED-ELISA relative to SN were 93.33% and 90.00%,respectively;rnPED-ELISA compared with TSZ PEDV antibody diagnosis Kit,91.67%concordance was obtained.200 serum samples were detected by this method,the total masculine ratio was69.5%.Therefore,this rnPED-ELISA based on recombinant truncated N protein antigen had good sensitivity and specificity,could afforded a simple and rapidmeans for assessment of vaccination in the field and investigation of PED epidemiology.