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Multiplex loop-mediated isothermal amplification (multi-LAMP) assay for rapid detection of mcr-1 to mcr-5 in colistin-resistant bacteria

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Zhong, Lan-Lan, Zhou, Qian, Tan, Cui-yan, Roberts, Adam, Ahmed, Mohamed Abd El-Gawad El-Sayed, Chen, Guanping, DaiMin, Min, Yang, Fan, Xia, Yong, Liao, Kang, Liang, Yingjian, Yang, Yongqiang, Feng, Siyuan, Zheng, Xiaobin and Tian, Guo-Bao (2019) 'Multiplex loop-mediated isothermal amplification (multi-LAMP) assay for rapid detection of mcr-1 to mcr-5 in colistin-resistant bacteria'. Infection and Drug Resistance, Vol 2019, Issue 12, pp. 1877-1887.

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Abstract

Purpose:
The discovery of the plasmid-mediated colistin resistancegenes, mcr,revealed a mechanism of transmission of colistin resistance, which is a major, globalpublic health concern especially among individuals infected with carbapenem-resistant Gram-negative bacteria. To monitor the spread and epidemiology of mcr genes, a convenient and reliable method to detect mcr genes in clinical isolates is needed, especially in the primary care institutions. This study aimed to establish a restriction endonuclease-based multiplex loop-mediated isothermal amplification (multi-LAMP) assay to detect mcr genes (mcr-1 to mcr-5) harbored by colistin-resistant bacteria.
Methods:
A triple-LAMP assay for mcr-1, mcr-3, and mcr-4 and a double-LAMP assay for mcr-2and mcr-5 were established. The sensitivity and specificity of the LAMP reactions were determined via electrophoresis and visual detection.
Results:
The sensitivity of the LAMP assay was 10-fold greater than that of PCR, with high specificity among the screened primers. Specific mcr genes were distinguished in accordance with band numbers and the fragment length of the digested LAMP amplification products. Furthermore, the LAMP assay was confirmed as a rapid and reliable diagnostic technique upon application for clinical samples, and the results were consistent with those of conventional PCR assay.
Conclusion:
The multi-LAMP assay is a potentially promising method to detect mcr genes and will, if implemented,help prevent infections by drug-resistant bacteria in primary-care hospitals due to rapid and reliable surveillance.To our knowledge, this is the first study to report the application of LAMP to detect mcr-2 to mcr-5genes and the first time that multi-LAMP has been applied to detect mcrgenes.

Item Type: Article
Subjects: QU Biochemistry > Genetics > QU 470 Genetic structures
QU Biochemistry > Genetics > QU 550 Genetic techniques. PCR. Chromosome mapping
QV Pharmacology > Anti-Bacterial Agents. Tissue Extracts > QV 350 Anti-bacterial agents (General or not elsewhere classified)
QW Microbiology and Immunology > QW 51 Morphology and variability of microorganisms. Microbial genetics.
Faculty: Department: Biological Sciences > Department of Tropical Disease Biology
Digital Object Identifer (DOI): https://doi.org/10.2147/IDR.S210226
Depositing User: Stacy Murtagh
Date Deposited: 17 Jul 2019 09:26
Last Modified: 30 Jul 2019 09:10
URI: https://archive.lstmed.ac.uk/id/eprint/11014

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