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目的了解温州地区不同来源蜡样芽胞杆菌毒力基因分布、生化分型和多位点序列分型(MLST)特征。方法参照GB 4789.14—2014对127株蜡样芽胞杆菌进行鉴定和生化分型,同时采用PCR方法检测10种毒力基因,并进行MLST基因分型。结果 127株蜡样芽胞杆菌同时携带Nhe A、Nhe B、Nhe C基因的菌株占94.5%(120/127),而同时携带hbl A、hbl C、hbl D基因的占9.4%(12/127),携带ces基因的占7.9%(10/127);127株菌株除10株不能进行生化分型外,其余117株分为2型(0.8%,1/127)、5型(3.9%,5/127)、8型(1.6%,2/127)、9型(63.8%,81/127)和10型(22.0%,28/127);通过MLST分析,72株菌株共分为28个ST序列型,ST26(18.1%,13/72)、ST144(15.3%,11/72)、ST92(6.9%,5/72)和ST164(6.9%,5/72)为常见ST型别。28个ST型聚类分析显示温州地区蜡样芽胞杆菌分为4个序列群和13个单态群。结论温州地区蜡样芽胞杆菌毒力基因携带率较高,遗传关系具有多样性。
Objective To understand the gene distribution, biochemical typing and multilocus sequence typing (MLST) of Bacillus cereus from different sources in Wenzhou area. Methods According to GB 4789.14-2014, 127 strains of Bacillus cereus were identified and biochemically typed. Ten virulence genes were detected by PCR and genotyped by MLST. Results The strains of Bacillus cereus carrying Nhe A, Nhe B and Nhe C genes accounted for 94.5% (120/127), while those carrying hbl A, hbl C and hbl D genes accounted for 9.4% (12/127) , 7.9% (10/127) carrying ces gene; 117 strains were divided into 2 types (0.8%, 1/127), 5 types (3.9%, 5 / 127), 8 (1.6%, 2/127), 9 (63.8%, 81/127) and 10 (22.0%, 28/127) ST26 (18.1%, 13/72), ST144 (15.3%, 11/72), ST92 (6.9%, 5/72) and ST164 (6.9%, 5/72) were the most common ST genotypes. 28 ST-type cluster analysis showed that Bacillus cereus in Wenzhou area is divided into four sequence groups and 13 singlet groups. Conclusion The Bacillus cereus virulence genes in Wenzhou have a higher carrying rate and genetic diversity.