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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 TSV RNA模板
A multiplex polymerase chain reaction (multiplex PCR) technique was developed for the simultaneous detection of white spot syndrome virus (WSSV) and taura virus (TSV). According to the sequences of WSSV and TSV, two pairs of primers complementary to the sequences of WSSV and TSV were designed and synthesized. Multiplex PCR amplification of WSSV DNA and TSV RNA templates in the same sample was carried out by using the two pairs of primers. Two specific PCR amplification bands of 30 6 bp (WSSV) and 2 31 bp (TSV) were obtained, which were consistent with the experimental design. However, the results of PCR amplification of other WSS pathogens were negative. The results of sensitivity test showed that, This multiplex PCR assay detected 10 pg of WSSV DNA and 1 pg of TSV RNA template