应用多重PCR检测鉴别对虾白斑综合征病毒和桃拉病毒  

Detection of White Spot Syndrome Virus and Taura Syndrome Virus of Penaeid Shrimp by Multiplex PCR

作　　者：谢芝勋[1] 庞耀珊[1] 何竞铭[1] 邓显文[1] 唐小飞[1] 刘加波[1] 谢志勤[1] 廖敏[1] 文雪 

机构地区：[1]广西壮族自治区兽医研究所,广西南宁530001 [2]广西壮族自治区海洋研究所,广西北海536000

出　　处：《中国兽医学报》2005年第1期13-15,共3页Chinese Journal of Veterinary Science

基　　金：广西科技攻关项目 (桂科攻 0 3 2 2 0 0 6-3 A)部分内容

摘　　要：研究建立了一种可同时检测对虾白斑综合征病毒 (WSSV)和桃拉病毒 (TSV)的多重聚合酶链式反应 (多重PCR)技术。根据 WSSV和 TSV基因序列 ,设计合成了 2对分别与 WSSV和 TSV某段基因序列互补的引物 ,用这 2对引物对同一样品中的 WSSV DNA和 TSV RNA模板进行多重 PCR扩增 ,结果均同时得到了 2条特异的与实验设计相符的 30 6 bp(WSSV)和 2 31bp(TSV)多重 PCR扩增带 ,而对其他对虾病病原的 PCR扩增结果均为阴性 ;敏感性测定结果表明 ,该多重 PCR技术能检出 10 pg的 WSSV DNA和 1pg的 TSVA multiplex polymerase chain reaction was optimized to simultaneously detect two pathogens of WSSV and TSV in this study.Two sets of specific primers were designed according to the sequences of WSSV and TSV gene fragement.It was shown that all samples which contained WSSV and TSV,could be amplified by the multiplex PCR using these two sets primers,yielding two specific bands of WSSV 306 bp and TSV 231 bp,but no specific band was amplified from other penaeid shrimp pathogenic virus and bacteria.As little as 10 pg of WSSV DNA and 1 pg of TSV RNA were detected using gel electrophoresis in this multiplex PCR.

关 键 词：WSSV 桃拉病 对虾白斑综合征病毒 引物 多重聚合酶 虾病 多重PCR检测 扩增 鉴别 RNA 

分 类 号：R378.21[医药卫生—病原生物学] S945[医药卫生—基础医学]