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Rapid Detection of Multi-Resistance Strains Carrying mcr-1 Gene Using Recombinase-Aided Amplification Directly on Clinical Samples

With the increasingly severe problem of bacterial resistance, colistin, as the last line of defense, has attracted attention again. Mobile colistin resistance (mcr-1) gene is involved in the horizontal transmission of colistin resistance in Gram-negative bacteria (GNB), which is a serious threat to...

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Autores principales: Fan, Zheng, Feng, Yanling, Xu, Wenjian, Feng, Junxia, Yan, Chao, Fu, Tongtong, Zhao, Hanqing, Cui, Jinghua, Gan, Lin, Liu, Shiyu, Du, Shuheng, Zhang, Rui, Xu, Ziying, Li, Nannan, Xue, Guanhua, Yuan, Jing
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9009391/
https://www.ncbi.nlm.nih.gov/pubmed/35432248
http://dx.doi.org/10.3389/fmicb.2022.852488
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author Fan, Zheng
Feng, Yanling
Xu, Wenjian
Feng, Junxia
Yan, Chao
Fu, Tongtong
Zhao, Hanqing
Cui, Jinghua
Gan, Lin
Liu, Shiyu
Du, Shuheng
Zhang, Rui
Xu, Ziying
Li, Nannan
Xue, Guanhua
Yuan, Jing
author_facet Fan, Zheng
Feng, Yanling
Xu, Wenjian
Feng, Junxia
Yan, Chao
Fu, Tongtong
Zhao, Hanqing
Cui, Jinghua
Gan, Lin
Liu, Shiyu
Du, Shuheng
Zhang, Rui
Xu, Ziying
Li, Nannan
Xue, Guanhua
Yuan, Jing
author_sort Fan, Zheng
collection PubMed
description With the increasingly severe problem of bacterial resistance, colistin, as the last line of defense, has attracted attention again. Mobile colistin resistance (mcr-1) gene is involved in the horizontal transmission of colistin resistance in Gram-negative bacteria (GNB), which is a serious threat to human health. Therefore, rapid detection of mcr-1 gene presence in clinical samples is crucial. In this study, a Recombinase-aided amplification(RAA) method for mcr-1 was successfully constructed, with sensitivity of 20 copies/reaction. In addition, amplification signal could only be detected in the strain containing mcr-1 gene among 14 different bacterial species. The method was then used to test a total of 672 clinical samples from a pediatric hospital in Beijing. Five strains harbored mcr-1 genes were isolated from mcr-1-positive clinical samples and identified as Escherichia coli. Multi-locus sequence typing (MLST) analysis showed that the five E. coli belonged to different ST types. Notably, the mcr-1 gene from the isolates could be transferred conjugately to the recipient strain E. coli J53, with highest transfer efficiency up to 57–58%, suggesting that the mcr-1 gene was located on the plasmid. These findings showed that the RAA assay has potential to be a rapid and sensitive mcr-1 gene screening test for clinical samples, and mcr-1 could be transmitted vertically and horizontally between and within bacterial species in a plasmid-mediated manner.
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spelling pubmed-90093912022-04-15 Rapid Detection of Multi-Resistance Strains Carrying mcr-1 Gene Using Recombinase-Aided Amplification Directly on Clinical Samples Fan, Zheng Feng, Yanling Xu, Wenjian Feng, Junxia Yan, Chao Fu, Tongtong Zhao, Hanqing Cui, Jinghua Gan, Lin Liu, Shiyu Du, Shuheng Zhang, Rui Xu, Ziying Li, Nannan Xue, Guanhua Yuan, Jing Front Microbiol Microbiology With the increasingly severe problem of bacterial resistance, colistin, as the last line of defense, has attracted attention again. Mobile colistin resistance (mcr-1) gene is involved in the horizontal transmission of colistin resistance in Gram-negative bacteria (GNB), which is a serious threat to human health. Therefore, rapid detection of mcr-1 gene presence in clinical samples is crucial. In this study, a Recombinase-aided amplification(RAA) method for mcr-1 was successfully constructed, with sensitivity of 20 copies/reaction. In addition, amplification signal could only be detected in the strain containing mcr-1 gene among 14 different bacterial species. The method was then used to test a total of 672 clinical samples from a pediatric hospital in Beijing. Five strains harbored mcr-1 genes were isolated from mcr-1-positive clinical samples and identified as Escherichia coli. Multi-locus sequence typing (MLST) analysis showed that the five E. coli belonged to different ST types. Notably, the mcr-1 gene from the isolates could be transferred conjugately to the recipient strain E. coli J53, with highest transfer efficiency up to 57–58%, suggesting that the mcr-1 gene was located on the plasmid. These findings showed that the RAA assay has potential to be a rapid and sensitive mcr-1 gene screening test for clinical samples, and mcr-1 could be transmitted vertically and horizontally between and within bacterial species in a plasmid-mediated manner. Frontiers Media S.A. 2022-03-31 /pmc/articles/PMC9009391/ /pubmed/35432248 http://dx.doi.org/10.3389/fmicb.2022.852488 Text en Copyright © 2022 Fan, Feng, Xu, Feng, Yan, Fu, Zhao, Cui, Gan, Liu, Du, Zhang, Xu, Li, Xue and Yuan. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Fan, Zheng
Feng, Yanling
Xu, Wenjian
Feng, Junxia
Yan, Chao
Fu, Tongtong
Zhao, Hanqing
Cui, Jinghua
Gan, Lin
Liu, Shiyu
Du, Shuheng
Zhang, Rui
Xu, Ziying
Li, Nannan
Xue, Guanhua
Yuan, Jing
Rapid Detection of Multi-Resistance Strains Carrying mcr-1 Gene Using Recombinase-Aided Amplification Directly on Clinical Samples
title Rapid Detection of Multi-Resistance Strains Carrying mcr-1 Gene Using Recombinase-Aided Amplification Directly on Clinical Samples
title_full Rapid Detection of Multi-Resistance Strains Carrying mcr-1 Gene Using Recombinase-Aided Amplification Directly on Clinical Samples
title_fullStr Rapid Detection of Multi-Resistance Strains Carrying mcr-1 Gene Using Recombinase-Aided Amplification Directly on Clinical Samples
title_full_unstemmed Rapid Detection of Multi-Resistance Strains Carrying mcr-1 Gene Using Recombinase-Aided Amplification Directly on Clinical Samples
title_short Rapid Detection of Multi-Resistance Strains Carrying mcr-1 Gene Using Recombinase-Aided Amplification Directly on Clinical Samples
title_sort rapid detection of multi-resistance strains carrying mcr-1 gene using recombinase-aided amplification directly on clinical samples
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9009391/
https://www.ncbi.nlm.nih.gov/pubmed/35432248
http://dx.doi.org/10.3389/fmicb.2022.852488
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