猪圆环病毒1型抗体IPMA检测方法的建立及应用  

Development and application of an immunoperoxidase monolayer assay for detection of antibodies against porcine circovirus type 1

作　　者：黄立平[1] 刘长明[1] 危艳武[1] 张朝霞[1] 陆月华[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：《中国兽医科学》2008年第9期738-742,共5页Chinese Veterinary Science

基　　金：国家引进国际先进农业科学技术计划(948)项目(2006-Z6);国家科技支撑计划项目(2006BAD06A07)

摘　　要：建立了一种用于检测猪圆环病毒1型(PCV1)血清抗体的免疫过氧化物酶单层细胞试验(IPMA)方法。对该方法的反应条件、特异性、敏感性及重复性等进行了系统试验,并与用昆虫杆状病毒表达的PCV1-Cap重组蛋白抗原建立的间接ELISA的检测结果进行了比较。结果显示,用IPMA法对已知PCV1和猪圆环病毒2型(PCV2)参考阳性血清进行检测,血清稀释度≥1∶100即可消除2种病毒间的低水平交叉反应,与其他几种猪病毒参考阳性血清无交叉反应;与用昆虫杆状病毒表达的PCV1-Cap重组蛋白抗原建立的间接ELISA的检测符合率为88.9%。用该方法对黑龙江省、吉林省、河北省、上海市、辽宁省等地猪场送检的257份血清样品进行检测,PCV1和PCV2血清抗体阳性检出率分别为19.1%和80.5%,PCV1抗体阳性猪中有95.9%为PCV2阳性,表明我国猪群已存在PCV1感染,在临床上PCV1与PCV2多以混合感染形式存在。建立的PCV1-IPMA方法具有操作简便、特异和敏感等特点,不失为PCV1流行病学调查及血清抗体检测的有效技术手段。An immunoperoxidase monolayer assay（IPMA） was developed for detection of antibodies against porcine cireovirus type 1 （PCV1）. The reaction conditions, specificity, sensitivity, and repeatability of IPMA were optimized, evaluated, and compared with those of ELISA based on the baculovirus-expressed PCV1 recombinant capsid protein as coating antigen. Different dilutions of PCV1 and PCV2 positive reference sera were assayed by IPMA,and the low level of cross-reaction of antibody against PCV1 with that against PCV2 could be eliminated by serum dilution of more than 1 ： 100. There was no cross-reaction with reference sera against other porcine viruses. In comparison,88.9% agreement rate was obtained between IPMA and ELISA. A total of 257 serum samples from Heilongjiang, Jilin, Hebei, Shanghai, Liaoning provinces were detected using PCVI-IPMA and PCV2qPMA. The positive rate was 19.1 % and 80.5 % for PCV1 and PCV2, respectively. There were antibodies against PCV2 in approximately 95. 9% of PCV1- positive samples, indicating that mixed infectious of PCV1 with PCV2 existed in the above-mentioned provinces. The results of comparison showed that the developed PCVI-IPMA offered a convenient tool for the epidemiological investigation of PCV1 infections and evaluation of antibodies against the virus.

关 键 词：猪圆环病毒1型 免疫过氧化物酶单层细胞试验 抗体检测 

分 类 号：S852.659.2[农业科学—基础兽医学] R446.61[农业科学—兽医学]