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根据GenBank上已发表链球菌cps2J毒力基因序列设计合成了一对可扩增长度为1 152 bp目的片段的引物,成功建立了特异性PCR方法用于食源性溶血链球菌毒力基因cps2J的检测。该方法的特异性和灵敏性试验结果显示,对大肠杆菌、沙门氏菌、金黄色葡萄球菌、霍乱弧菌、副溶血性弧菌、绿脓杆菌、粪肠杆菌、变形杆菌以及藤黄微球菌的PCR检测结果均呈阴性,对分离的9株食源性溶血链球菌菌株进行了检测,电泳结果均呈阳性;检测目的菌株DNA的最低敏感度可达3.2×10~(-1)ng/m L。结果表明,此法特异性强,灵敏性高,能准确地检测肉制品中携带毒力基因cps2J的溶血链球菌,可作为食源性溶血链球菌快速诊断和流行病学调查的手段。
According to the published sequences of cps2J virulence genes of Streptococcus in GenBank, a pair of primers was designed to amplify the target fragment of 1 152 bp in length. A specific PCR method was successfully established for the virulence gene cps2J of S. hemolyticus Detection. Specificity and sensitivity of the method The results of the test showed that PCR against Escherichia coli, Salmonella, Staphylococcus aureus, Vibrio cholerae, Vibrio parahemolyticus, Pseudomonas aeruginosa, Enterobacter faecalis, Proteus and Micrococcus luteus The test results were negative. The 9 strains of isolated hemolytic Streptococcus were tested and the results of electrophoresis were all positive. The lowest susceptibility of the tested strains to DNA was 3.2 × 10 ~ (-1) ng / m L . The results show that this method is highly specific and sensitive, and can accurately detect hemolytic streptococcus carrying cps2J in meat products, which can be used as a rapid diagnosis and epidemiological investigation of food-borne hemolytic streptococcus.