禽四种病毒多重PCR诊断技术的建立和应用  

Establishment and Application of a Multiplex PCR Diagnostic TecHnology for Four Virus

作　　者：张琳[1,2] 胡北侠[1,2] 杨少华[1,2] 许传田[1,2] 陈正涛[1,3] 郝明飞[1,3] 张秀美[1,2] 

机构地区：[1]山东省畜禽疫病防治与繁育重点实验室,山东济南250100 [2]山东省农业科学院畜牧兽医研究所,山东济南250100 [3]山东农业大学动物科技学院,山东泰安271018

出　　处：《家畜生态学报》2011年第4期71-74,共4页Journal of Domestic Animal Ecology

基　　金：现代农业产业技术体系建设专项资金

摘　　要：根据禽呼肠孤病毒(ARV)、禽流感病毒(AIV)、新城疫病毒(NDV)和鸡传染性支气管炎病毒(IBV)的基因保守区设计了4对特异性引物,建立了针对四种病毒的多重PCR检测体系,并对该体系进行了条件优化及特异性、敏感性试验。该体系扩增的四种病毒基因片断大小分别为199bp(ARV)、264bp(AIV)、362bp(NDV)和459bp(IBV),且特异性、敏感性良好,能够检出1pg AIV和10pg ARV、NDV、IBV的RNA。临床应用结果证明,该体系具有很高的实用价值和应用前景。According to the conserved region sequence of avian reovirus（ARV）, avian influenza virus （AIV）, Newcastle disease virus（NDV） and infectious bronchitis virus（IBV）, four pairs of specific primers were designed to develop a multiplex PCR. Then the multiplex PCR was optimized and the specificity and sensitivity were tested. The gene fragments of 199 bp（ARV）, 264 bp（AIV）, 362 bp（NDV）, 459 bp （IBV） were amplified from the RNA mixture which contained even little as 1 pg RNA of AIV and 10 pg RNA of ARV, NDV and IBV. The clinical samples detection showed that the developed multiplex PCR could rapidly and simultaneously diagnose ARV, AIV, NDV and IBV in chicken.

关 键 词：多重PCR 禽呼肠孤病毒 禽流感病毒 新城疫病毒 鸡传染性支气管炎病毒 

分 类 号：S811.5[农业科学—畜牧学]