PRV-IPMA抗体检测试剂盒的研制及其应用

Development and application of immunoperoxidase monolayer assay kit for detection of antibody against porcine pseudorabies virus

本研究建立了一种免疫过氧化物酶单层细胞试验法(IPMA),用于猪伪狂犬病病毒(PRV)血清抗体检测。通过对IPMA反应条件的优化,组装成PRV-IPMA诊断试剂盒,并对该试剂盒检测的敏感性、特异性、重复性及保存期等进行了试验。结果表明,IPMA检测相对于SN的敏感性为96.2%,特异性为97.7%,两者的总符合率为96.9%;该试剂盒检测的重复性好,与其它病毒参考血清无交叉反应;试剂盒可在-20℃保存12个月,用该试剂盒检测PRV人工感染猪血清,于感染后2周抗体全部阳转,健康对照组猪血清抗体检测均为阴性结果。对来自黑龙江、吉林、上海、内蒙古、河北等地采集的5个猪场后备母猪150份血清样本进行检测,PRV血清抗体阳性检出率为16.7%～50%。检测结果表明这些猪场后备猪群仍需加强疫苗免疫。该试剂盒的研制为我国PRV流行病学调查和疫苗免疫效果的评价提供了技术手段。

A method for detecting antibodies of porcine pseudorabies virus （PRV） was developed by immunoperoxidase monolayer assay （IPMA）, and assembled as PRV-IPMA kit. Compared with the SN, the sensitivity of IPMA was 96.2 %, the specificity was 97.7 %, and the Coincident rate was 96.9 %. Repeatability and preservation of the kit were evaluated and the results showed that the PRV-IPMA kit was stable for 12 months stored at -20 ℃. There was no cross-reaction with other reference sera of porcine viruses. About 100 % positive rate could be detected by the assay for PRV-infected pig sera at the 2nd week post-inoculation. All sera of negative control pigs were detected to be negative. PRV antibody positive rate ranged from 16.7 % to 50 % through the survey of 150 serum samples from five farm replacement gilts, which collected from Heilongjiang, Jilin, Shanghai, Inner Mongolia and Hebei provinces. The PRV-IPMA kit offers a convenient tool for PRV epidemiology survey and evaluation of vaccine efficiency.