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

Autor: Chenze Lu, Jingwen Wang, Leiming Pan, Xiuying Gu, Wenjing Lu, Di Chen, Cen Zhang, Qin Ye, Chaogeng Xiao, Pengpeng Liu, Yulong Tang, Biao Tang, Guangrong Huang, Jiehong Fang, Han Jiang
Jazyk: angličtina
Rok vydání: 2023
Předmět:
Zdroj: Frontiers in Microbiology, Vol 13 (2023)
Druh dokumentu: article
ISSN: 1664-302X
DOI: 10.3389/fmicb.2022.1062577
Popis: 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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