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Establishment and application of recombinase polymerase amplification combined with a lateral flow dipstick for the detection of mcr-1 in uncultured clinical samples.
- Source :
-
International journal of antimicrobial agents [Int J Antimicrob Agents] 2024 May; Vol. 63 (5), pp. 107140. Date of Electronic Publication: 2024 Mar 13. - Publication Year :
- 2024
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Abstract
- 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.<br />Methods: A rapid detection technique combining recombinase polymerase amplification with a lateral flow dipstick targeting uncultured clinical samples was developed.<br />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.<br />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.<br /> (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)
- Subjects :
- Recombinases
Humans
Feces microbiology
DNA-Directed DNA Polymerase
Colistin pharmacology
Anti-Bacterial Agents pharmacology
Nucleic Acid Amplification Techniques
Enterobacteriaceae drug effects
Enterobacteriaceae genetics
Enterobacteriaceae isolation & purification
Ethanolaminephosphotransferase analysis
Ethanolaminephosphotransferase genetics
Bacterial Proteins analysis
Bacterial Proteins genetics
Drug Resistance, Bacterial genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1872-7913
- Volume :
- 63
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- International journal of antimicrobial agents
- Publication Type :
- Academic Journal
- Accession number :
- 38490574
- Full Text :
- https://doi.org/10.1016/j.ijantimicag.2024.107140