猪繁殖与呼吸综合征病毒荧光微球抗体检测方法的建立

Establishment of a Fluorescent Microsphere Immunoassay Assay for Detection of Porcine Reproductive and Respiratory Syndrome Virus

为建立一种荧光微球免疫层析方法用于猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV)抗体检测,将兔抗羊二抗和重组N蛋白分别作为质控线(C线)和检测线(T线),喷涂于硝酸纤维素膜上,通过优化反应条件,建立了PRRSV荧光微球抗体检测方法,采用ROC曲线确定本方法的判定临界值,并对其敏感性、特异性、重复性及符合率进行了分析。结果显示:本研究建立的PRRSV荧光微球抗体检测方法能够检测到32倍稀释的阳性血清,对猪伪狂犬病病毒、猪瘟病毒、猪细小病毒和猪圆环病毒2型阳性血清的检测结果均为阴性;重复性试验结果的变异系数为3.71%~8.99%;与美国爱德士试剂盒的符合率为91.30%,具有较高的一致性。结果表明,本研究建立的PRRSV荧光微球抗体检测方法性能稳定、结果可靠,为PRRSV抗体快速检测提供了新方法。

In order to establish a fluorescence microsphere immunoassay assay for detection of antibodies against porcine reproductive and respiratory syndrome virus(PRRSV),rabbit anti-sheep IgG(HL)and recombinant N protein were sprayed onto nitrocellulocellulose membranes as the quality control line(C line)and detection line(T line),respectively.After optimization of reaction conditions,a fluorescence microsphere immunoassay assay for detection of PRRSV antibodies was established,and the critical value was determined by the ROC curve,then its sensitivity,specificity,repeatability and coincidence rate were evaluated and analyzed.The results showed that the established method could be used to detect 32 times diluted positive serum,and negative to react with the positive serum of porcine pseudorabies virus(PRV),classical swine fever virus(CSFV),porcine parvovirus(PPV)and porcine circovirus type 2(PCV-2);the variable coefficient of repeatability test was 3.71%to 8.99%;and the coincidence rate with IDEXX kit was 91.30%,indicating a high consistency.In conclusion,a new tool was provided for rapid detection of antibodies against PRRSV by the established method with stable performance and reliable results.