等温多自配引发扩增技术快速检测学校餐饮中三种食源性致病菌  

Rapid Detection of Three Foodborne Pathogenic Bacteria in School Catering by Isothermal Multiple Self-matching-initiated Amplification Technique

作　　者：焦强 陈万胜 周楠 张庆发 刘智勇 JIAO Qiang;CHEN Wansheng;ZHOU Nan;ZHANG Qingfa;LIU Zhiyong(Henan Province Food Inspection Research Institute,Key Laboratory of Food Safety Quick Testing and Smart Supervision Technology for State Regulation,Zhengzhou 450000,China;Guangzhou DI’AO Bio-technology Co.,Ltd.,Guangzhou 510507,China)

机构地区：[1]河南省食品检验研究院,国家市场监管重点实验室(食品安全快速检测与智慧监管技术),河南郑州450000 [2]广州迪澳生物科技有限公司,广东广州510507

出　　处：《食品工业科技》2023年第18期371-377,共7页Science and Technology of Food Industry

基　　金：2021年度市场监管总局科技计划项目(2021MK065)。

摘　　要：目的:本文研究了一种适合学校餐饮中快速鉴别三种食源性致病菌的检测方法。方法:基于等温多自配引发扩增技术(isothermal multiple self-matching-initiated amplification,IMSA),选择沙门氏菌的invA基因、大肠埃希氏菌O157:H7/NM的rfbE基因和单核细胞增生李斯特氏菌的prfA基因设计特异性引物,检测菌液灵敏度、特异性、最短增菌时间、最低含菌量检出限等指标;以食品安全国家标准食品微生物学检验(GB/T 4789.4-2016、GB/T 4789.30-2016、GB/T 4789.36-2016)为参考方法,比较两种方法的一致性。结果:该方法对沙门氏菌、大肠埃希氏菌O157:H7/NM和单核细胞增生李斯特氏菌的菌液灵敏度分别为2.14×10^(3)、2.79×10^(3)和3.62×10^(3)CFU/mL;特异性高达100%;人工污染最短的增菌时间分别为6、8和8 h;最低含菌量检出限分别为2.14、2.79和3.62 CFU/25 g;74例食品样本的IMSA法结果与国标法一致性达100%。结论:IMSA技术检测食源性致病菌的灵敏度高、特异性强、结果准确,可在短时间内完成食品中三种致病菌的检测,适合于学校餐饮中致病菌的快速鉴别。Objective:In this paper,a detection method suitable for rapid identification of three pathogenic bacteria from food in the school catering was studied.Method:Based on isothermal multiple self-matching-initiated amplification technology(IMSA),strain-specific primers were designed for Salmonella invA gene,Escherichia coli O157:H7/NM rfbE gene and Listeria monocytogenes prfA gene to test the sensitivity,specificity,shortest enrichment time,and minimum detection limit of bacterial content.Food microbiology inspection according to National Food Safety Standards(GB/T 4789.4-2016,GB/T 4789.30-2016,GB/T 4789.36-2016)were the reference methods,and compared the consistency of the two methods.Result:The results showed that the sensitivity of our method to Salmonella,Escherichia coli O157:H7/NM and Listeria monocytogenes were 2.14×10^(3),2.79×10^(3)and 3.62×10^(3)CFU/mL respectively,the specificity was 100%,the shortest enrichment time of artificial contamination were 6,8 and 8 hours respectively.The detection limits of the lowest bacteria content were 2.14,2.79 and 3.62 CFU/25 g respectively,and the consistency of the results of 74 food samples using IMSA method and the reference method was 100%.Conclusion:IMSA technology of pathogenic bacteria detection in food had the advantage of having high sensitivity,strong specificity and accurate results,which could complete the detection of three pathogenic bacteria in food in a short time and was suitable for the rapid identification of pathogenic bacteria in school catering.

关 键 词：沙门氏菌 大肠埃希氏菌 O157:H7/NM 单核细胞增生李斯特氏菌 等温多自配引发扩增技术 

分 类 号：TS201.3[轻工技术与工程—食品科学]