多重PCR检测无公害畜禽肉和水产品中4种致病菌

建立无公害畜禽肉和水产品中肠出血性大肠杆菌(EHEC)、沙门氏菌、副溶血性弧菌(VP)和单核细胞增生性李斯特氏菌(LM)的多重PCR检测方法,为这些致病菌的快速诊断提供实验依据。选择分别针对EHEC溶血素基因hlyAB、副溶血性弧菌属保守序列toxR基因、沙门氏菌侵袭基因invA和LM的iap基因特异的4对引物,先分别进行单重PCR扩增,再同时加入4对引物进行多重PCR扩增,扩增产物经测序验证。建立的多重PCR方法可简便、快速、灵敏地实现对EHEC、LM、沙门氏菌和VP的同时检测,在畜禽肉和水产品中的检测灵敏度达到10^3cfu/mL。

Simultaneous Detection of Common Food-borne Bacterial Pathogens in Innocuous Poultry and Seafood by Multiplex PCR Assay

To develop a multiplex PCR assay to detect Enterohemorrhagic Escherichia coli (EHEC), Vibrio parahaemolyticus (VP), Salmonella spp. and Listeria monocytogenes(LM) in innocuous poulthy and seafood simultaneously, four pairs of primers were designed from hemolysis gene (hlyAB) of EHEC, toxR gene of VP, invA gene of Salmonella spp. and iap gene of LM. They were detected by conventional PCR and multiplex PCR assay. DNA sequencing was used to examine PCR products. This multiplex PCR method was a highly specific, rapid and sensitive method which could simultaneously detect EHEC, VP, Salmonella spp. and LM. The detection level was 10~(3)cfu/mL.