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目的建立一种基于细菌RNA聚合酶β亚基编码基因(rpo B)的克罗诺杆菌种鉴定PCR方法,对不同来源的克罗诺杆菌进行种水平鉴定。方法通过rpo B基因设计针对克罗诺杆菌7个种的引物,以9株参考菌株作为参照,对261株2012—2014年收集的我国婴儿配方食品和谷基辅食类食品及临床腹泻病例来源的克罗诺杆菌进行种水平鉴定。结果 9株参考菌株PCR结果与对应大小一致,261株克罗诺杆菌中的179株为阪崎克罗诺杆菌,56株为丙二酸盐克罗诺杆菌,13株为尤尼沃斯克罗诺杆菌,11株为都柏林克罗诺杆菌,2株为苏黎世克罗诺杆菌,分别占总菌株数的68.58%、21.46%、4.98%、4.21%和0.77%。结论建立的克罗诺杆菌PCR法种水平鉴定具有特异、便捷、灵敏等优势,可为我国食品安全克罗诺杆菌的风险监测和控制提供技术支持。
OBJECTIVE: To establish a method for identification of Cronobacter species based on the rpo B gene of the bacterial RNA polymerase subunit, and to identify species of Cronobacter from different sources. Methods The primers of seven species of Cronobacter were designed according to the rpo B gene. Nine strains of reference strains were used as reference. 261 strains of infant formula and cereal-based food and clinical diarrhea collected in 2012-2014 Cronobacter species identification. Results The PCR results of nine reference strains were consistent with those of the corresponding strains. Among the 261 strains of Cronobacteria, 179 strains were Crossessia kosakii, 56 strains were Corynebacterium malonate and 13 strains were Univaspor Bacillus nocturnalis, 11 strains were Corynebacterium clones and 2 strains were Corynebacterium zuriensis, accounting for 68.58%, 21.46%, 4.98%, 4.21% and 0.77% of the total strains, respectively. Conclusion The identification of Cronobacter pylori PCR method has the advantages of specificity, convenience and sensitivity. It can provide technical support for the risk monitoring and control of Corynebacterium fumigatus in China.