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

作　　者：张超范[1] 刘长明[1] 危艳武[1] 田志军[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室猪传染病研究室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2008年第6期445-449,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家"948"计划(2006-Z6);国家科技支撑计划项目(2006BAD06A07)

摘　　要：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.

关 键 词：猪伪狂犬痛病毒 免疫过氧化物酶单层细胞试验 抗体检测试剂盒 

分 类 号：S852.43[农业科学—基础兽医学]