水貂肠炎病毒双抗体夹心ELISA检测方法的建立  

Establishment of double antibody sandwich ELISA for detection of mink enteritis virus

作　　者：王建科[1] 程世鹏[1] 易立[1] 杨莘[1] 罗彬[1] 许红丽[1] 闫喜军[1] 武华[1] 

机构地区：[1]中国农业科学院特产研究所,吉林吉林132109

出　　处：《中国兽医科学》2011年第2期183-187,共5页Chinese Veterinary Science

基　　金：吉林省科技发展计划项目(20080213;20090531)

摘　　要：以抗水貂肠炎病毒(mink enteritis virus,MEV)单克隆抗体为捕获抗体、兔抗MEV多克隆抗体为检测抗体,建立了MEV双抗体夹心ELISA检测方法。该方法用于MEV抗原的检测。经过试验,单克隆抗体的最适稀释度为1∶20,兔抗MEV多克隆抗体的最适稀释度为1∶3 200。用该ELISA方法分别检测MEV、水貂阿留申病病毒、犬瘟热病毒样品。结果表明,ELISA方法具有良好的特异性。用该ELISA和PCR同时检测158份临床样品,其中ELISA检测40份为阳性,PCR检测44份为阳性,该ELISA的特异性和敏感性分别为95.6%和79.5%,这2种方法的符合率为91.1%。该方法的建立为MEV的检测及水貂病毒性肠炎的流行病学调查提供了工具。In order to establish double antibody sandwich enzyme-linked immunosorbent assay（DAS-ELISA） for detection of mink enteritis virus（MEV）,anti-MEV monoclonal antibody and rabbit anti-MEV polyclonal antibody were used as the capture antibody and detecting antibody,respectively.The optimal dilution of the capture antibody and the detecting antibody capable of detecting MEV antigens were 1∶20 and 1∶3 200 in the check-board titration respectively.Positive samples with MEV,Aleutian mink disease virus and canine distemper virus were examined by the established ELISA respectively.In result,the deve-loped DAS-ELISA had good specificity.A total of 158 samples were tested both by the developed DAS-ELISA and by polymerase chain reaction（PCR）.40 of the tested samples were positive by the developed ADS-ELISA and 44 by PCR.The specificity and sensitivity of the developed DAS-ELISA were 95.6% and 79.5%,respectively.The coincidental rate between the developed DAS-ELISA and PCR was 91.1%.These results indicated that the developed DAS-ELISA was suitable for rapid detection and epidemiological investigation of MEV infection in mink.

关 键 词：水貂肠炎病毒 酶联免疫吸附试验 检测 单克隆抗体 

分 类 号：S852.659.2[农业科学—基础兽医学]