PRRSV-IPMA抗体检测试剂盒的研制及其应用

Application and development of immunoperoxidase monolayer assay kit for detection of antibody against porcine reproductive and respiratory syndrome virus

建立了一种检测猪生殖与呼吸综合征病毒（PRRSV）血清抗体的免疫过氧化物酶单层细胞试验法（IPMA），并组装成PRRSV-IPMA抗体检测试剂盒。对该试剂盒的敏感性、特异性、重复性及保存期等进行了试验，并与国外IDEXX ELISA试剂盒进行了比较。结果显示，用该试剂盒检测PRRSV人工接种猪血清样品，第1周抗体阳性检出率为50％，从第2周起抗体阳性检出率均为100％，而对照组猪血清抗体检测均为阴性。该试剂盒重复性好，-20℃保存18个月稳定，与其他病毒参考血清无交叉反应，与国外IDEXX ELISA试剂盒的符合率为83．3%。用该试剂盒对采自黑龙江、吉林、辽宁、河北、上海等地13个猪场的520份正常猪血清样品进行了抗体检测，结果，4个猪场阳性率高达90％以上，仅有2个猪场为阴性，其余猪场阳性率为10％～57%。

A kit for detection of antibody against porcine reproductive and respiratory syndrome virus （PRRSV） was prepared by immunoperoxidase monolayer assay（IPMA） and assembled as PRRSV-IPMA kit. Sensitivity, specificity, repeatability and preservation of the kit were evaluated. Using the PRRSVIPMA kit sera from PRRSV-infected pigs were detected to be 50% and 100% of positive rates at week 1 and 2 post-inoculation, respectively. The sera from control group pigs showed negative reaction. In comparison, parallel detections were done using the PRRSV-IPMA kit and the commercial PRRSV-ELISA kit from IDEXX Co. , and the agreement rate was 83.3%. The PRRSV-IPMA kit was stable for 18 months at -20℃, and didn＇t cross-react with the other virus reference sera. PRRSV antibodies in 520 serum samples from 13 farms of Heilongjiang, Jilin, Liaoning, Hebei and Shanghai were detected by the PRRSV- IPMA kit. The positive rates of four farms were more than 90%, and only two farms were negative, and 10%-57% in the rest of farms.