Virulence gene detection and antimicrobial susceptibility analysis of Bacillus cereus fresh food isolates
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摘要: 目的:研究生鲜食品中蜡样芽孢杆菌的毒力基因及耐药性。方法:在成都市周边农贸市场和路边小摊采集各种生鲜食品共100份,采用MYP选择性培养基初步分离蜡样芽孢杆菌菌株,采用16S rRNA序列比对分析鉴定得到蜡样芽孢杆菌分离菌株,采用PCR检测分离鉴定菌株中蜡样芽孢杆菌13个毒力基因的携带情况,进一步采用纸片扩散法检测菌株对18种抗生素的耐药性。结果:从100份生鲜食品样本中,检出蜡样芽孢杆菌24株,检出率为24%,其中,路边小摊的检出率(70%)高于农贸市场(18.9%)。呕吐型基因ces和cer的检出率较低,仅在1株中发现;腹泻型基因nheC有24株检出;cytK有13株检出;bceT有12株检出;hblA有11株检出;nheB有10株检出;hblC、hblD、nheA各有9株检出;hly-Ⅱ有7株检出;hblB、entFM检出率为0;蜡样芽孢杆菌的4个看家基因rpoB、gyrB、groEL、vrrA在24株分离菌株中检出率为100%。同时发现,生鲜食品来源的蜡样芽孢杆菌分离菌株对杆菌肽B、磺胺甲亚唑、苯唑西林、青霉素表现出较高耐药性,耐药率大于96%;而对阿米卡星、氯霉素、庆大霉素、亚胺培南耐药率小于7%。结论:本研究通过分离鉴定生鲜食品源蜡样芽孢杆菌,研究其毒力基因携带及耐药性,为进一步评价生鲜食品中蜡样芽孢杆菌的安全风险提供参考数据。Abstract: Objective:To isolate and identify Bacillus cereus in food and further investigate the virulence genes and antimicrobial resistance of Bacillus cereus fresh food isolates.Methods:100 fresh food products samples were collected from food markets and food stands of Chengdu,respectively.Then,Bacillus cereus strains were identified by MYP selective medium. 16S rRNA sequencing analysis was used to further identify Bacillus cereus species. Virulence genes of Bacillus cereus were detected by PCR method. The susceptibility of Bacillus cereus to 18 antibiotics was detected by disk diffusion method. Results:24 strains of Bacillus cereus fresh food isolates were detected from 100 fresh food samples,the isolation rate was 24%.The detection rates of Bacillus cereus from food stands samples were 70%,much higher than that from food markets samples(18.9%). Among all the 13 virulence genes,both ces and cer were only found to be occurred in one isolate,with a very low detection rate,nheC occurred in 24 isolates,cytK occurred in 13 isolates,bceT occurred in 12 isolates,hblA occurred in 11 isolates,nheB occurred in 10 isolates,hblC,nheA and hblD occurred in 9 isolates,hly-Ⅱ occurred in 7 isolates,hblB and entFM were not found,the detection rates for4 housekeeping genes(rpoB,gyrB,groEL and vrrA)were100%. Bacillus cereus fresh food isolates showed high resistance to bacitracin B,sulfamethoxazole,oxacillin and penicillin,and the resistant rate was more than 96%. For amikacin,chloramphenicol,gentamicin,and amphenol,the resistance rates were less than 7%. Conclusion:The results would lay a foundation for further assessing the risk of Bacillus cereus in fresh food.
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[1] Bioengineering
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