全基因组测序技术用于深圳地区一起食物中毒中金黄色葡萄球菌遗传特征分析

doi:10.12183/j.scjpm.2024.1104

摘要/Abstract

摘要： 目的对深圳地区发生的一起金黄色葡萄球菌（简称金葡菌）食物中毒菌株进行基因特征分析,了解金葡菌的肠毒素类型,对引起食物中毒致病菌来源进行溯源。方法采集食物中毒病例呕吐物、肛拭子、可疑食品和环境涂抹样本,对菌株进一步分离和生化鉴定,用荧光PCR方法检测肠毒素基因,ELISA方法检测肠毒素,用微量肉汤稀释法对分离到的金葡菌进行药敏试验。用全基因组测序方法对菌株进行测序,将菌株序列与Resfinder、CARD、MLST和VFDB数据库比对,分别获得耐药基因、ST型和毒力因子。利用生物信息分析平台绘制系统发育树。结果 13份样品中检出金葡菌,其中病例肛拭子2份,呕吐物1份,食品5份,环境涂抹样5份,样品阳性率65%。阳性样本中共挑取可疑菌落36株菌,均未检出sea、seb、sec、sed及see肠毒素基因,ELISA方法检出24份样品产E型肠毒素,VFDB数据库结果显示36株菌携带新型肠毒素基因seg和类肠毒素基因selm、seln、selo。所有菌株对青霉素耐药（100%）,对其他抗生素敏感（0%）。携带β-内酰胺类耐药基因的菌株表现出100%的表型耐药,而携带四环素类、喹诺酮类、氨基糖苷类等耐药基因的菌株没有表现出相应的表型耐药。36株菌均为ST-1281型,菌株的SNP突变位点差距较小,菌株间SNP突变位点矩阵为0~13位。结论本次事件是由ST-1281型金葡菌感染引起,而且是较为少见的新型肠毒素引起的食物中毒。

关键词: 金黄色葡萄球菌, 肠毒素, 全基因组测序, 微量肉汤稀释法, ST型

Abstract: Objective To analyze the genetic characteristics of a food poisoning strain of Staphylococcus aureus (SA) in Shenzhen and understand the types of enterotoxins of SA, and to trace the source of the pathogenic bacteria causing by food poisoning. Methods Vomitus, anal swabs, suspicious foods, and environmental smear samples of the food poisoning were collected, and the strains were further isolated and biochemically identified. The enterotoxin genes were detected by fluorescence PCR, enterotoxins were detected by ELISA, and the isolated SA were tested by microbroth dilution. The whole genome sequencing method was used to sequence the strains, and the strain sequences were compared with the Resfinder, CARD, MLST, and VFDB databases to obtain drug resistance genes, ST types, and virulence factors, respectively. The phylogenetic tree was drawn by the bioinformatics analysis platform. Results SA was detected in 13 samples, with a positive rate of 65%, including 2 anal swabs from patients, 1 vomit sample, 5 food samples, and 5 environmental smear samples. A total of 36 strains of suspicious colonies were selected from the positive samples, none of them were detected with sea, seb, sec, sed, and see enterotoxin genes, 24 samples were detected by ELISA to produce type E enterotoxin, and the VFDB database results showed that 36 strains of SA carried the novel enterotoxin gene seg and enterotoxoid genes selm, seln, and selo. All strains were resistant to penicillin (100%) and sensitive to other antibiotics (0%). The strains carrying β-lactam resistance genes showed 100% phenotypic resistance, while the strains carrying tetracycline, quinolones, aminoglycosides, and other resistance genes did not show corresponding phenotypic resistance. All 36 strains were ST-1281 type, the SNP mutation site gap of the strains was small, and the SNP mutation site matrix ranging from 0 to 13. Conclusion This incident was caused by ST-1281 type SA, and it is a relatively rare food poisoning caused by a new type of enterotoxin.

Key words: Staphylococcus aureus, Enterotoxin, Whole genome sequencing, Microbroth dilution method, ST type

中图分类号:

R155

何娇明, 杨东芳, 马佳智, 何欣儿, 鞠长燕. 全基因组测序技术用于深圳地区一起食物中毒中金黄色葡萄球菌遗传特征分析[J]. 华南预防医学, 2024, 50(12): 1104-1108.

HE Jiaoming, YANG Dongfang, MA Jiazhi, HE Xiner, JU Changyan. Whole genome sequencing for the analysis of the genetic characteristics of Staphylococcus aureus in a food poisoning of Shenzhen[J]. South China Journal of Preventive Medicine, 2024, 50(12): 1104-1108.

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