应用多重PCR检测鉴别对虾白斑综合征病毒和桃拉病毒

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306bp(WSSV)和231bp(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10pg的WSSV DNA和1pg的TSV RNA模板。

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

A 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.