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为了研究副溶血弧菌双重PCR检测方法,试验采用tlh和toxR基因序列设计2对特异性引物进行双重PCR检测,扩增出450 bp和368 bp目的片段。结果表明:在同一PCR反应体系中仅副溶血弧菌可同时扩增出2种基因片段,4株对照菌无任何扩增条带;该双重PCR检测方法最低能检测1.660 7×103cfu/mL菌体浓度的副溶血弧菌。说明试验所建立的双重PCR检测方法特异性强、敏感性高、方法简单、用时短,可用于副溶血弧菌引起的水产动物疾病的诊断与分子流行病学调查。
In order to study the method of dual PCR detection of Vibrio parahaemolyticus, two pairs of specific primers were designed for double PCR detection using tlh and toxR gene sequences, and 450 bp and 368 bp fragments were amplified. The results showed that only two Vibrio parahaemolyticus strains could amplify two kinds of gene fragments in the same PCR reaction system without any amplification bands. The double PCR method could detect 1.660 7 × 103 cfu / mL bacteria Body concentration of Vibrio parahaemolyticus. It shows that the double PCR method established by the experiment is specific, sensitive, simple, and short in time. It can be used in the diagnosis and molecular epidemiological investigation of aquatic animal diseases caused by Vibrio parahaemolyticus.