Detection assay of polymyxin resistance coding mcr-1 gene based on CRISPR/Cas13a system
IntroductionPolymyxins are reserved as an ultimate defense against multidrug-resistant bacteria. The emergence of the polymyxin resistance gene mcr-1 poses a potential risk for the treatment of severe infections caused by Gram-negative bacteria. Timely detection and monitoring the mcr-1 gene are ess...
Saved in:
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2025-06-01
|
| Series: | Frontiers in Cellular and Infection Microbiology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fcimb.2025.1553681/full |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849471301862817792 |
|---|---|
| author | Yingjie Song Qiang Hu Yao Han Hongbo Liu Zhenyang Huang Mengwei Niu Xue Dong Kuocheng Yan Li Jin Hao Li Yansong Sun |
| author_facet | Yingjie Song Qiang Hu Yao Han Hongbo Liu Zhenyang Huang Mengwei Niu Xue Dong Kuocheng Yan Li Jin Hao Li Yansong Sun |
| author_sort | Yingjie Song |
| collection | DOAJ |
| description | IntroductionPolymyxins are reserved as an ultimate defense against multidrug-resistant bacteria. The emergence of the polymyxin resistance gene mcr-1 poses a potential risk for the treatment of severe infections caused by Gram-negative bacteria. Timely detection and monitoring the mcr-1 gene are essential for guiding anti-infective therapy and controlling the spread of polymyxin resistance. Quantitative real-time PCR (qPCR) is one of the common methods for detecting resistance genes. However, qPCR has equipment dependency, and is not feasible in primary healthcare settings. Currently, there remains a lack of a highly sensitive and portable method for detecting the mcr-1 gene.MethodsWe established and optimized detection assays of the mcr-1 gene based on CRISPR/Cas13a system and lateral flow strips. The detection method was preliminarily evaluated using clinical isolates from Escherichia coli, compared with qPCR.ResultsThe method for detecting the mcr-1 gene based on the CRISPR/Cas13a system and lateral flow strips was established, with a detection limit of 100 copies/mL. This method demonstrated high analytical specificity, with no cross-reactivity detected in non-mcr-1 and non-resistant strains. Among 36 clinical isolates, the method identified 31 strains as positive for the mcr-1 gene, and had a 100% concordance rate with the results of qPCR.ConclusionsWe established a detection method for the polymyxin resistance mcr-1 gene based on the CRISPR/Cas13a system. This method enables visual readouts without instruments, making it potentially applicable to primary healthcare settings and field surveillance. |
| format | Article |
| id | doaj-art-e17180d224b54d3daad2ca8ecc04c389 |
| institution | Kabale University |
| issn | 2235-2988 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular and Infection Microbiology |
| spelling | doaj-art-e17180d224b54d3daad2ca8ecc04c3892025-08-20T03:24:52ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882025-06-011510.3389/fcimb.2025.15536811553681Detection assay of polymyxin resistance coding mcr-1 gene based on CRISPR/Cas13a systemYingjie Song0Qiang Hu1Yao Han2Hongbo Liu3Zhenyang Huang4Mengwei Niu5Xue Dong6Kuocheng Yan7Li Jin8Hao Li9Yansong Sun10State Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, Beijing, ChinaState Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, Beijing, ChinaState Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, Beijing, ChinaChinese PLA Center for Disease Control and Prevention, Beijing, ChinaState Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, Beijing, ChinaState Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, Beijing, ChinaState Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, Beijing, ChinaState Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, Beijing, ChinaState Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, Beijing, ChinaState Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, Beijing, ChinaState Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, Beijing, ChinaIntroductionPolymyxins are reserved as an ultimate defense against multidrug-resistant bacteria. The emergence of the polymyxin resistance gene mcr-1 poses a potential risk for the treatment of severe infections caused by Gram-negative bacteria. Timely detection and monitoring the mcr-1 gene are essential for guiding anti-infective therapy and controlling the spread of polymyxin resistance. Quantitative real-time PCR (qPCR) is one of the common methods for detecting resistance genes. However, qPCR has equipment dependency, and is not feasible in primary healthcare settings. Currently, there remains a lack of a highly sensitive and portable method for detecting the mcr-1 gene.MethodsWe established and optimized detection assays of the mcr-1 gene based on CRISPR/Cas13a system and lateral flow strips. The detection method was preliminarily evaluated using clinical isolates from Escherichia coli, compared with qPCR.ResultsThe method for detecting the mcr-1 gene based on the CRISPR/Cas13a system and lateral flow strips was established, with a detection limit of 100 copies/mL. This method demonstrated high analytical specificity, with no cross-reactivity detected in non-mcr-1 and non-resistant strains. Among 36 clinical isolates, the method identified 31 strains as positive for the mcr-1 gene, and had a 100% concordance rate with the results of qPCR.ConclusionsWe established a detection method for the polymyxin resistance mcr-1 gene based on the CRISPR/Cas13a system. This method enables visual readouts without instruments, making it potentially applicable to primary healthcare settings and field surveillance.https://www.frontiersin.org/articles/10.3389/fcimb.2025.1553681/fullmcr-1polymyxinantibiotic resistanceCRISPR/Cas13alateral flow strip |
| spellingShingle | Yingjie Song Qiang Hu Yao Han Hongbo Liu Zhenyang Huang Mengwei Niu Xue Dong Kuocheng Yan Li Jin Hao Li Yansong Sun Detection assay of polymyxin resistance coding mcr-1 gene based on CRISPR/Cas13a system Frontiers in Cellular and Infection Microbiology mcr-1 polymyxin antibiotic resistance CRISPR/Cas13a lateral flow strip |
| title | Detection assay of polymyxin resistance coding mcr-1 gene based on CRISPR/Cas13a system |
| title_full | Detection assay of polymyxin resistance coding mcr-1 gene based on CRISPR/Cas13a system |
| title_fullStr | Detection assay of polymyxin resistance coding mcr-1 gene based on CRISPR/Cas13a system |
| title_full_unstemmed | Detection assay of polymyxin resistance coding mcr-1 gene based on CRISPR/Cas13a system |
| title_short | Detection assay of polymyxin resistance coding mcr-1 gene based on CRISPR/Cas13a system |
| title_sort | detection assay of polymyxin resistance coding mcr 1 gene based on crispr cas13a system |
| topic | mcr-1 polymyxin antibiotic resistance CRISPR/Cas13a lateral flow strip |
| url | https://www.frontiersin.org/articles/10.3389/fcimb.2025.1553681/full |
| work_keys_str_mv | AT yingjiesong detectionassayofpolymyxinresistancecodingmcr1genebasedoncrisprcas13asystem AT qianghu detectionassayofpolymyxinresistancecodingmcr1genebasedoncrisprcas13asystem AT yaohan detectionassayofpolymyxinresistancecodingmcr1genebasedoncrisprcas13asystem AT hongboliu detectionassayofpolymyxinresistancecodingmcr1genebasedoncrisprcas13asystem AT zhenyanghuang detectionassayofpolymyxinresistancecodingmcr1genebasedoncrisprcas13asystem AT mengweiniu detectionassayofpolymyxinresistancecodingmcr1genebasedoncrisprcas13asystem AT xuedong detectionassayofpolymyxinresistancecodingmcr1genebasedoncrisprcas13asystem AT kuochengyan detectionassayofpolymyxinresistancecodingmcr1genebasedoncrisprcas13asystem AT lijin detectionassayofpolymyxinresistancecodingmcr1genebasedoncrisprcas13asystem AT haoli detectionassayofpolymyxinresistancecodingmcr1genebasedoncrisprcas13asystem AT yansongsun detectionassayofpolymyxinresistancecodingmcr1genebasedoncrisprcas13asystem |