【摘要】 目的建立一种快速、简便、准确,适合于基层应用的检测鉴定Lm的方法。方法选取iap基因作为靶序列设计一对引物,用该引物对4株标准株及4株无关菌进行PCR扩增,得到进一步验证。采用PCR和常规方法同时对人工感染的牛奶进行检测鉴定。结果4株标准株获得了预期700bp左右的扩增片段,而同属的L.welshimeri、L.grayi、粪肠球菌和蜡样芽孢杆菌均没有这一特异性条带。检测人工感染牛奶20份,检出率为90%,高于传统分离培养法(60%)。结论建立的PCR法可用于单增李斯特氏菌的快速、特异、敏感诊断。更多还原

【Abstract】 Objective To establish a quick,convenient and concise testing method adaptable for detection of Listeria monocytogenes(Lm)by using PCR technology.Methods A pair of primers were designed by taking the iap gene as objective sequences and using these primers to amplify the genes of 6 reference Lm and 4 reference bacteria.The sequences of bacteria in artificial infected milk were identified after amplification by PCR and conventional method and the results were compared.Results Expected 700bp fragments were obtained from 4 reference strains,while no such specific fragments were identified in the L.Weshimeri,L.grayi,Enterococcus faecalis and B.dereus after amplification.90% of the 20 artificially infected milk samples were detected by PCR,higher than that of the conventional one(60%).Conclusion The established PCR technique can be used for quick,specific,sensitive diagnosis of Listeria monocytogenes in MJME.更多还原