食品与发酵工业 >

2023 , Vol. 49 >Issue 22: 279 - 286

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.035031

分析与检测

基于双重DARQ-LAMP方法同时检测单增李斯特菌和金黄色葡萄球菌

  • 陈清莹 ,
  • 刘丹 ,
  • 张杏果 ,
  • 韦锦源 ,
  • 钟青萍
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  • (广东省食品质量安全重点实验室,华南农业大学 食品学院,广东 广州,510642)
第一作者：硕士研究生(钟青萍教授为通信作者,E-mail:zhongqp@scau.edu.cn)

收稿日期: 2023-02-06

  修回日期: 2023-02-20

  网络出版日期: 2023-12-25

基金资助

广东省基础与应用基础研究基金项目(2021A1515011083);国家自然科学基金面上项目(31972046);国家重点研发计划重点专项(2017YFC1601203);广东省科技计划项目(2020B1212060059)

收起

Simultaneous detection of Listeria monocytogenes and Staphylococcus aureus based on dual DARQ-LAMP method

  • CHEN Qingying ,
  • LIU Dan ,
  • ZHANG Xingguo ,
  • WEI Jinyuan ,
  • ZHONG Qingping
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  • (Guangdong Provincial Key Laboratory of Food Quality and Safety, College of Food Science, South China Agricultural University, Guangzhou 510642, China)

Received date: 2023-02-06

  Revised date: 2023-02-20

  Online published: 2023-12-25

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摘要

环介导等温扩增技术(loop-mediated isothermal amplification,LAMP)是快速检测食源性致病菌的常用方法。该文分别针对单增李斯特菌和金黄色葡萄球菌的特异性基因hlyA和nuc设计引物,通过优化反应条件,建立了一种可同时检测单增李斯特菌和金黄色葡萄球菌的基于淬灭基团释放环介导等温扩增检测方法(detection of amplification by release of quenching-LAMP,DARQ-LAMP),并评估了该方法的特异性和灵敏度。结果表明该方法可在60 min内完成对单增李斯特菌和金黄色葡萄球菌的同时检测,检测限分别为7.3×102 copies/mL和2.3×102 copies/mL,特异性良好、灵敏度高,在实际样品检测中检测限为2.8×104 CFU/mL和2.4×104 CFU/mL。该双重DARQ-LAMP方法能同时检测2种食源性致病菌,缩短检测时间,提高了检测效率,为多重LAMP检测食源性致病菌的研究提供了新的思路。

关键词： 食源性致病菌; 单增李斯特菌; 金黄色葡萄球菌; 基于淬灭基团释放环介导等温扩增技术; 快速检测

本文引用格式

陈清莹 , 刘丹 , 张杏果 , 韦锦源 , 钟青萍 . 基于双重DARQ-LAMP方法同时检测单增李斯特菌和金黄色葡萄球菌[J]. 食品与发酵工业, 2023 , 49(22) : 279 -286 . DOI: 10.13995/j.cnki.11-1802/ts.035031

Abstract

Loop-mediated isothermal amplification (LAMP) is an effective method for rapid detection of foodborne pathogens. In this study, DARQ-LAMP (detection of amplification by release of quenching and loop-mediated isothermal amplification) method was established for simultaneous detection of Listeria monocytogenes and Staphylococcus aureus. The primers were designed for the specific genes hlyA of L. monocytogenes and nuc of S. aureus, respectively, and the reaction conditions were optimized. The specificity and sensitivity of the method were also evaluated. Results showed that L. monocytogenes and S. aureus could be detected simultaneously in 60 minutes, and the detection limits were 7.3×102 and 2.3×102 copies/mL, respectively. The method was of good specificity and sensitivity, and the detection limits of 2.8×104 and 2.4×104 CFU/mL could be achieved in real samples. This dual DARQ-LAMP assay could detect two kinds of foodborne pathogens simultaneously, which would shorten the detection time and improve the detection efficiency, providing a new idea for the study of multiple LAMP detection of foodborne pathogens.

Key words： foodborne pathogens; Listeria monocytogenes; Staphylococcus aureus; detection of amplification by release of quenching and loop-mediated isothermal amplification (DARQ-LAMP); rapid detection

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