金黄色葡萄球菌食物中毒的多重PCR快速检测及产肠毒素耐药分析

Rapid detection of food-borne disease caused by Staphylococcus aureus by multiplex PCR and analysis of enterotoxin resistance

目的对一起疑似金黄色葡萄球菌肠毒素引起的食物中毒事件进行实验室检测和鉴定。方法采集7份环境涂抹物,4份食品和1份病例样本,提取样品总DNA进行14种食源性致病菌的多重PCR检测,对样品进行葡萄球菌肠毒素的ELISA检测。同时按照《食品安全国家标准食品卫生微生物学检验》进行金黄色葡萄球菌的分离鉴定,分离得到菌株进行耐药分析和肠毒素基因分型(SEA-SEE)。结果多重PCR和ELISA检测台面涂抹物和病例样本中金黄色葡萄球菌和肠毒素均为阳性。分离出3株和10株金黄色葡萄球菌,分别携带肠毒素基因sea和seb。其中台面涂抹物和病例样本都有携带sea毒素基因的分离株,携带同种肠毒素基因的菌株耐药结果一致,所有分离株对氨苄西林和青霉素耐药。结论该起食物中毒由金黄色葡萄球菌肠毒素引起,并存在耐药现象,相关部门应加强对小型饭馆的卫生监督管理。

Objective A suspected food borne disease caused by Staphylococcus aureus enterotoxin were conducted for laboratory testing and identification. Methods Seven environmental smears,four copoies of food and one case sample were collected. The total DNA of the samples was extracted for the detection of 14 food-borne pathogens by multiplex PCR. Staphylococcal enterotoxins were detected in the samples by enzyme-linked immunosorbent assay（ ELISA） directly. S. aureus was isolated and identified based on the The National Standards for Food Safety Microbiological Examination of Food Hygiene. The isolated strains were conducted for the classification of enterotoxin gene by polymerase chain reaction（ SEA-SEE）,and antimicrobial resistance experiment was performed on the isolates. Results Multiplex PCR and ELISA results showed both S. aureus and enterotoxin were positive in operate table smear and case sample. 3 and 10 strains of S. aureus were respectively isolated and harbored enterotoxin genes of sea and seb. Among them,the operate table smear and case sample isolates carry sea enterotoxin gene at the same time. The drug susceptibility results of the S. aureus isolates carrying same enterotoxin genes were consistent,which were resistant to ampicillin and penicillin. Conclusion The food-borne diseases was caused by Staphylococcus aureus enterotoxin,in which the antimicrobial resistance exist,so the hygiene supervision and management of small restaurants should be strengthened by the relevant departments.