双重PCR快速检测副溶血弧菌

Rapid detection of Vibrio parahaemolyticus with Duplex-PCR

目的:建立一种快速、敏感、特异的双重PCR方法检测副溶血弧菌。方法:根据GenBank收录的副溶血弧菌tdh、trh、tlh基因序列用vtII软件分别设计引物,应用多重PCR技术同时扩增副溶血弧菌的特异性基因tdh、trh、tlh。结果:副溶血弧菌标准株和实验菌株均能扩增一条或两条目的带,扩增产物tdh、trh、tlh片段大小分别为224,466,381 bp。结论:tdh、trh基因分别存在于不同副溶血弧菌株中,而tlh基因在不同副溶血弧菌中都广泛存在,具有种属特异性,因此可以用来特异快速的检测副溶血弧菌。

Objective：To develop a Duplex-PCR for the rapid,sensitive and specific detection of Vibrio parahaemolyticus.Methods：With primers designed according to the gene sequences of tdh,trh and tlh published in GenBank,Duplex-PCR was applied to amplify the genes of tdh,trh and tlh of Vibrio parahaemolyticus.Results：Products of tdh gene,trh gene and tlh gene of Vibrio parahaemolyticus strains with duplex PCR amplification was of one or two Electrophoresis strips.The amplified fragment tdh,trh and tlh was 224,466 and 381 bp.Conclusion：The tdh gene and the trh gene are in different Vibrio parahaemolyticus stains.While the tlh gene is widely in Vibrio parahaemolyticus and has genus characteristics so we can amplify the tdh gene as a rapid detection method for Vibrio parahaemolyticus.