Establishment and application of recombinase polymerase amplification combined with a lateral flow dipstick for the detection of mcr-1 in uncultured clinical samples

作者：Jun Feng, Zhen Xu, Yuan Zhuang, Jiayuan Luo, Yong Chen, Yitong Wu, Jiayi Fei, Mingxiang Liu, Jiahui Xia, Jing Zhang, Meihua Liu, Xie Xiaohong, Zhengan Yuan, Min Chen · 发表于：International Journal of Antimicrobial Agents · 年份：2024 · DOI：10.1016/j.ijantimicag.2024.107140 · 被引用次数：6 · 研究领域：Antibiotic Resistance in Bacteria、Escherichia coli research studies、Biosensors and Analytical Detection

OBJECTIVES: The rapid dissemination of the mcr-1 gene via plasmid-mediated transfer has raised concerns regarding the efficacy of colistin as a last-resort treatment for multidrug-resistant Gram-negative bacterial infections. Current mcr-1 gene detection methods mainly focus on cultured bacteria, which is a complex and time-consuming process requiring skilled personnel, making it unsuitable for field analysis. METHODS: A rapid detection technique combining recombinase polymerase amplification with a lateral flow dipstick targeting uncultured clinical samples was developed. RESULTS: This new method targeting the mcr-1 gene region (23 232-23 642 bp, no. KP347127.1) achieved a low detection limit of 10 copies/μL. The whole process was carried out with high specificity and was completed within 20 min. The evaluation assay was conducted using 45 human faecal samples; 16 strains yielded a 98% accuracy, closely matching antimicrobial susceptibility outcomes. CONCLUSIONS: The novel method integrates nucleic acid extraction, isothermal amplification, and a test assay, suggesting the potential for timely colistin resistance surveillance in frontline disease control and healthcare settings, supporting future prevention and clinical standardization efforts.