基于高分辨率溶解曲线分析鉴别食品中的3种李斯特氏菌

Identification of Three Listeria Species in Foods based on High Resolution Melting Curve Analysis

本研究利用高分辨率熔解曲线的方法,以iap基因为靶标新设计一对引物同时鉴别单增李斯特氏菌、伊氏李斯特氏菌和英诺克李斯特氏菌,其余14种常见食源性病原微生物扩增为阴性结果,单增李斯特氏菌和伊氏李斯特氏菌的检出限为10个拷贝,英诺克李斯特氏菌的检测限为50个拷贝。本研究对78份样品进行HRM-realtimePCR法、GB4789.30-2016和SN/T1870-2016(荧光PCR法)的检测,并对3种方法的检测结果进行统计学分析。结果显示:HRM法和国标方法、HRM法和荧光PCR法的检测结果之间存在统计学差异,国标方法和荧光PCR法检测结果之间无统计学差异。本研究建立的基于HRM-real time PCR法检测3种李斯特氏菌的方法快速高效、特异性好、成本低,适用于食品中李斯特氏菌的日常检测和监管。

In this study,a new pair of primers was designed to identify Listeria monocytogenes,Listeria evanescens and Listeria innominata at the same time with iap gene as the target by using high-resolution fusion curve method.The amplification of the other 14 common foodborne pathogens was negative.The detection limit of Listeria monocytogenes and Listeria evanescens was 10 copies,and that of Listeria innominata was 50 copies.In this study,78 samples were tested by HRM-real time PCR method,GB4789.30-2016 and SN/T 1870-2016(real-time PCR),and the results of three methods were analyzed statistically.The results showed that there were statistical differences between the detection results of HRM method and national standard method,HRM method and fluorescent PCR method,and there was no statistical difference between the results of national standard method and fluorescent PCR method.The method based on HRM-real time PCR for the detection of three Listeria species in this study is fast,efficient,specific,and low cost.It is suitable for the daily detection and supervision of Listeria species in food.