Rapid detection of multiple resistance genes to last-resort antibiotics in Enterobacteriaceae pathogens by recombinase polymerase amplification combined with lateral flow dipstick.
Rapid detection of multiple resistance genes to last-resort antibiotics in Enterobacteriaceae pathogens by recombinase polymerase amplification combined with lateral flow dipstick.
复制标题
重组酶聚合酶扩增结合侧流试纸快速检测肠杆菌科病原体对最后手段抗生素的多种耐药基因
DOI：
10.3389/fmicb.2022.1062577
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发表时间：
2022
影响因子：
5.2
通讯作者：
Jiang, Han
中科院分区：
文献类型：
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作者：
Lu, Chenze;Wang, Jingwen;Pan, Leiming;Gu, Xiuying;Lu, Wenjing;Chen, Di;Zhang, Cen;Ye, Qin;Xiao, Chaogeng;Liu, Pengpeng;Tang, Yulong;Tang, Biao;Huang, Guangrong;Fang, Jiehong;Jiang, Han
关键词：
在整个食物链和相关环境中的食用动物和人类群体中出现了令人担忧的对最后手段抗生素的多重耐药基因，这在世界范围内得到了越来越多的报道。肠杆菌科病原体被认为是此类抗生素耐药基因（ARG）的最常见宿主。因此，迫切需要一种快速、高效、准确的检测方法来同时筛查和监测肠杆菌科病原菌中的ARGs。我们的研究开发了重组酶聚合酶扩增（RPA）检测结合侧流试纸条（LFD），用于同时检测肠杆菌科病原体对最后手段抗生素的主要耐药基因，包括mcr-1，blaNDM-1和泰特（X4）。在37°C下40 min内完成粗DNA提取、扩增和阅读的整个过程，mcr-1、blaNDM-1和泰特（X4）的检测限为101 copies/μl。灵敏度分析显示，使用试纸条读数器，颜色信号与肠杆菌科病原体中mcr-1、blaNDM-1和泰特（X4）基因的模板浓度存在明显相关性（R2分别= 0.9881、R2 = 0.9745和R2 = 0.9807），允许使用多重RPA-LFD检测进行定量检测。因此，RPA-LFD检测方法可以适当地帮助检测食源性病原体中的多种对最后手段抗生素的耐药基因，并具有潜在的应用前景。
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, blaNDM-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°C, and the detection limit is 101 copies/μl for mcr-1, blaNDM-1 and tet(X4). Sensitivity analysis showed obvious association of color signals with the template concentrations of mcr-1, blaNDM-1 and tet(X4) genes in Enterobacteriaceae pathogens using a test strip reader (R2 = 0.9881, R2 = 0.9745, and R2 = 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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影响因子：
6.1
作者：
Cavalera S;Colitti B;Rosati S;Ferrara G;Bertolotti L;Nogarol C;Guiotto C;Cagnazzo C;Denina M;Fagioli F;Di Nardo F;Chiarello M;Baggiani C;Anfossi L
通讯作者：
Anfossi L
影响因子：
5.2
作者：
Li, Jiali;Ma, Biao;Zhang, Mingzhou
通讯作者：
Zhang, Mingzhou
影响因子：
3.7
作者：
Lillis L;Lehman D;Singhal MC;Cantera J;Singleton J;Labarre P;Toyama A;Piepenburg O;Parker M;Wood R;Overbaugh J;Boyle DS
通讯作者：
Boyle DS
影响因子：
3.7
作者：
通讯作者：
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影响因子：
1.6
作者：
通讯作者：
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