Luminex x-TAG技术在金黄色葡萄球菌肠毒素快速分型中的应用  

A rapid screening system for the detection of Staphylococcal enterotoxin based on Luminex x-TAG technology

作　　者：叶硕[1] 郑健 周伟艳[1] 高红[1] YE Shuo;ZHENG Jian;ZHOU Wei-yan;GAO Hong(Microbiology Laboratory,Ningbo Municipal Center for Disease Control and Prevention,Ningbo,Zhejiang 315010,China)

机构地区：[1]宁波市疾病预防控制中心微生物检验所,浙江宁波315010

出　　处：《中国卫生检验杂志》2020年第21期2564-2567,2571,共5页Chinese Journal of Health Laboratory Technology

基　　金：浙江省医药卫生科技计划项目(2017KY602);宁波市科技惠民项目(2017C50047)。

摘　　要：目的基于Luminex悬浮芯片系统,建立多种金黄色葡萄球菌肠毒素快速筛查体系。方法菌株来源为本中心2012年-2018年食品污染物检测中分离的158株金黄色葡萄球菌。分别设计金黄色葡萄球菌的通用引物及SEA、SEB、SEC、SED、SEE 5种特异性肠毒素分型引物构建成多重引物体系。并通过Luminex200平台进行分析。结果本体系能特异性鉴别金黄色葡萄球菌并对5种肠毒素进行分型,检出限达到103cfu/ml。实际菌株样品5种肠毒素基因总检出率为43.8%(70/160),sea阳性菌株最多(17.5%,28/160)。共发现16种肠毒素基因组合,sea(15株)、sec(9株)、sea+sec(8株)3种基因组合居于前三位。结论本液相芯片方法快速分型了本地区2012年-2018年食品污染物检测中分离的158株金黄色葡萄球菌,发现SEA型仍然是地区优势的毒力因子,为食品风险的监测和食物中毒的诊断提供有效保障。Objective To establish a rapid screening system based on Luminex suspension chip technology for the detection of Staphylococcal enterotoxins.Methods A total of 158 strains of Staphylococcus aureus used in this study were isolated from the surveillance of food pollutant in Ningbo from 2012 to 2018.Universal primers for the identification of Staphylococcus aureus and specific primers for the typing of SEs were designed.Luminex200 system was used for the analysis.Results Our system could specifically identify Staphylococcus aureus and typing five kinds of SEs.The detection limits could reach to 103 cfu/ml.The total detection rate of the SEs was 43.8%(70/160),while strains carrying sea accounting for the most(17.5%,28/160).A total of 16 SEs combinations were found,while sea(15 strains),sec(9 strains)and sea+sec(8 strains)were the top three.Conclusion 158 strains of Staphylococcus aureus were screened by our rapid typing system during this study.And we found SEA still the dominant virulence factor of Staphylococcus aureus in Ningbo.It is believed that our system would perform well in food safety risk surveillance and diagnosis of food poisoning after further improvement.

关 键 词：液相芯片 金黄色葡萄球菌肠毒素 LUMINEX 多重PCR 

分 类 号：R446.5[医药卫生—诊断学]