Rapid detection of multiple resistance genes to last-resort antibiotics in Enterobacteriaceae pathogens by recombinase polymerase amplification combined with lateral flow dipstick

文献类型： 外文期刊

第一作者： Lu, Chenze

作者： Lu, Chenze;Wang, Jingwen;Huang, Guangrong;Fang, Jiehong;Jiang, Han;Pan, Leiming;Gu, Xiuying;Lu, Wenjing;Chen, Di;Zhang, Cen;Ye, Qin;Xiao, Chaogeng;Liu, Pengpeng;Tang, Yulong;Tang, Biao;Tang, Biao

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关键词： recombinase polymerase amplification; lateral flow dipstick; Enterobacteriaceae; rapid detection; resistance genes to last-resort antibiotics

期刊名称：FRONTIERS IN MICROBIOLOGY （ 影响因子：5.2； 五年影响因子：6.2 ）

ISSN：

年卷期： 2023 年 13 卷

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收录情况： SCI

摘要： The worrying emergence of multiple resistance genes to last-resort antibiotics in food animals and human populations throughout the food chain and relevant environments has been increasingly reported worldwide. Enterobacteriaceae pathogens are considered the most common reservoirs of such antibiotic resistance genes (ARGs). Thus, a rapid, efficient and accurate detection method to simultaneously screen and monitor such ARGs in Enterobacteriaceae pathogens has become an urgent need. Our study developed a recombinase polymerase amplification (RPA) assay combined with a lateral flow dipstick (LFD) for simultaneously detecting predominant resistance genes to last-resort antibiotics of Enterobacteriaceae pathogens, including mcr-1, bla(NDM-1) and tet(X4). It is allowed to complete the entire process, including crude DNA extraction, amplification as well as reading, within 40 min at 37 & DEG;C, and the detection limit is 10(1) copies/mu l for mcr-1, bla(NDM-1) and tet(X4). Sensitivity analysis showed obvious association of color signals with the template concentrations of mcr-1, bla(NDM-1) and tet(X4) genes in Enterobacteriaceae pathogens using a test strip reader (R-2 = 0.9881, R-2 = 0.9745, and R-2 = 0.9807, respectively), allowing for quantitative detection using multiplex RPA-LFD assays. Therefore, the RPA-LFD assay can suitably help to detect multiple resistance genes to last-resort antibiotics in foodborne pathogens and has potential applications in the field.

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