猪圆环病毒2型免疫过氧化物酶单层细胞试验抗体检测试剂盒的研制及应用

Development and application of an immunoperoxidase monolayer assay based kit for detection of porcine circovirus type 2

本研究建立了一种免疫过氧化物酶单层细胞试验(Immunoperoxidase monolayer assay,IPMA),用于猪圆环病毒2型血清抗体检测,通过对IPMA反应条件的优化,组装了诊断试剂盒。研究结果表明,用IPMA检测猪圆环病毒2型人工感染猪血清,于感染后3周抗体阳转,第3周～10周抗体阳性检出率为92.8%(52/56),对照组猪血清抗体检测均为阴性(33/33)。试剂盒在-20℃稳定保存18个月与其他几种猪病毒参考血清无交叉反应,与用重组蛋白抗原建立的rcELISA符合率为89.2%。对来自黑龙江、吉林、河北、上海、内蒙古、云南、江西等地猪场健康成年猪血清480份和发病猪血清424份进行了检测,抗体检出率分别为91.7%和79.2%,表明我国猪群中猪圆环病毒2型污染相当严重。该试剂盒的研制为我国PCV2流行病学调查和疫苗免疫效果的评价提供了技术手段。

An immunoperoxidase monolayer assay （IPMA） based diagnostic kit was developed for antibody detection of porcine circovirus type 2 （PCV2） under optimized conditions This kit could detected 92.8 % （52/56） positive sera from PCV2-infected pigs 3 to 10 weeks post-inoculation, and all control pigs were detected negative. There was no cross-reaction with reference sera against other porcine viruses. The results showed 89.2 % consistency with that of rcELISA using recombinant protein as antigen. A tootal of 480 sera from normal pig herds and 424 sera from PMWS-infected pig herds collected from Heilongjiang, Jilin, Hebei, Shanghai, Inner Mongolia, Yunnan, Jiangxi in China were tested. PCV2 antibody was detected in 91.7 % sera of normal pig herds and 79.2 % from the PMWS-infected pig herds. The results showed that these pig herds were highlycontaminated with PCV2. The IPMA kit offers a convenient tool for PCV2 epidemiology survey and evaluation of vaccine efficiency.