Rapid culture‐independent loop‐mediated isothermal amplification detection of antimicrobial resistance markers from environmental water samples
Abstract Environmental water is considered one of the main vehicles for the transmission of antimicrobial resistance (AMR), posing an increasing threat to humans and animals health. Continuous efforts are being made to eliminate AMR; however, the detection of AMR pathogens from water samples often r...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2023-05-01
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| Series: | Microbial Biotechnology |
| Online Access: | https://doi.org/10.1111/1751-7915.14227 |
| _version_ | 1797839405183926272 |
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| author | Marwa M. Hassan Arnoud H. M. vanVliet Owen Higgins Liam P. Burke Alexandra Chueiri Louise O'Connor Dearbháile Morris Terry J. Smith Roberto M. La Ragione |
| author_facet | Marwa M. Hassan Arnoud H. M. vanVliet Owen Higgins Liam P. Burke Alexandra Chueiri Louise O'Connor Dearbháile Morris Terry J. Smith Roberto M. La Ragione |
| author_sort | Marwa M. Hassan |
| collection | DOAJ |
| description | Abstract Environmental water is considered one of the main vehicles for the transmission of antimicrobial resistance (AMR), posing an increasing threat to humans and animals health. Continuous efforts are being made to eliminate AMR; however, the detection of AMR pathogens from water samples often requires at least one culture step, which is time‐consuming and can limit sensitivity. In this study, we employed comparative genomics to identify the prevalence of AMR genes within among: Escherichia coli, Klebsiella, Salmonella enterica and Acinetobacter, using publicly available genomes. The mcr‐1, blaKPC (KPC‐1 to KPC‐4 alleles), blaOXA‐48, blaOXA‐23 and blaVIM (VIM‐1 and VIM‐2 alleles) genes are of great medical and veterinary significance, thus were selected as targets for the development of isothermal loop‐mediated amplification (LAMP) detection assays. We also developed a rapid and sensitive sample preparation method for an integrated culture‐independent LAMP‐based detection from water samples. The developed assays successfully detected the five AMR gene markers from pond water within 1 h and were 100% sensitive and specific with a detection limit of 0.0625 μg/mL and 10 cfu/mL for genomic DNA and spiked bacterial cells, respectively. The integrated detection can be easily implemented in resource‐limited areas to enhance One Health AMR surveillances and improve diagnostics. |
| first_indexed | 2024-04-09T15:57:26Z |
| format | Article |
| id | doaj.art-896ff65799f64d7eab991536d3eb958c |
| institution | Directory Open Access Journal |
| issn | 1751-7915 |
| language | English |
| last_indexed | 2024-04-09T15:57:26Z |
| publishDate | 2023-05-01 |
| publisher | Wiley |
| record_format | Article |
| series | Microbial Biotechnology |
| spelling | doaj.art-896ff65799f64d7eab991536d3eb958c2023-04-25T14:53:17ZengWileyMicrobial Biotechnology1751-79152023-05-0116597798910.1111/1751-7915.14227Rapid culture‐independent loop‐mediated isothermal amplification detection of antimicrobial resistance markers from environmental water samplesMarwa M. Hassan0Arnoud H. M. vanVliet1Owen Higgins2Liam P. Burke3Alexandra Chueiri4Louise O'Connor5Dearbháile Morris6Terry J. Smith7Roberto M. La Ragione8Department of Comparative Biomedical Sciences, School of Veterinary Medicine, Faculty of Health and Medical Sciences University of Surrey Guildford UKDepartment of Comparative Biomedical Sciences, School of Veterinary Medicine, Faculty of Health and Medical Sciences University of Surrey Guildford UKMolecular Diagnostics Research Group, School of Biological and Chemical Sciences University of Galway Galway IrelandAntimicrobial Resistance and Microbial Ecology Group, School of Medicine University of Galway Galway IrelandAntimicrobial Resistance and Microbial Ecology Group, School of Medicine University of Galway Galway IrelandAntimicrobial Resistance and Microbial Ecology Group, School of Medicine University of Galway Galway IrelandAntimicrobial Resistance and Microbial Ecology Group, School of Medicine University of Galway Galway IrelandMolecular Diagnostics Research Group, School of Biological and Chemical Sciences University of Galway Galway IrelandDepartment of Comparative Biomedical Sciences, School of Veterinary Medicine, Faculty of Health and Medical Sciences University of Surrey Guildford UKAbstract Environmental water is considered one of the main vehicles for the transmission of antimicrobial resistance (AMR), posing an increasing threat to humans and animals health. Continuous efforts are being made to eliminate AMR; however, the detection of AMR pathogens from water samples often requires at least one culture step, which is time‐consuming and can limit sensitivity. In this study, we employed comparative genomics to identify the prevalence of AMR genes within among: Escherichia coli, Klebsiella, Salmonella enterica and Acinetobacter, using publicly available genomes. The mcr‐1, blaKPC (KPC‐1 to KPC‐4 alleles), blaOXA‐48, blaOXA‐23 and blaVIM (VIM‐1 and VIM‐2 alleles) genes are of great medical and veterinary significance, thus were selected as targets for the development of isothermal loop‐mediated amplification (LAMP) detection assays. We also developed a rapid and sensitive sample preparation method for an integrated culture‐independent LAMP‐based detection from water samples. The developed assays successfully detected the five AMR gene markers from pond water within 1 h and were 100% sensitive and specific with a detection limit of 0.0625 μg/mL and 10 cfu/mL for genomic DNA and spiked bacterial cells, respectively. The integrated detection can be easily implemented in resource‐limited areas to enhance One Health AMR surveillances and improve diagnostics.https://doi.org/10.1111/1751-7915.14227 |
| spellingShingle | Marwa M. Hassan Arnoud H. M. vanVliet Owen Higgins Liam P. Burke Alexandra Chueiri Louise O'Connor Dearbháile Morris Terry J. Smith Roberto M. La Ragione Rapid culture‐independent loop‐mediated isothermal amplification detection of antimicrobial resistance markers from environmental water samples Microbial Biotechnology |
| title | Rapid culture‐independent loop‐mediated isothermal amplification detection of antimicrobial resistance markers from environmental water samples |
| title_full | Rapid culture‐independent loop‐mediated isothermal amplification detection of antimicrobial resistance markers from environmental water samples |
| title_fullStr | Rapid culture‐independent loop‐mediated isothermal amplification detection of antimicrobial resistance markers from environmental water samples |
| title_full_unstemmed | Rapid culture‐independent loop‐mediated isothermal amplification detection of antimicrobial resistance markers from environmental water samples |
| title_short | Rapid culture‐independent loop‐mediated isothermal amplification detection of antimicrobial resistance markers from environmental water samples |
| title_sort | rapid culture independent loop mediated isothermal amplification detection of antimicrobial resistance markers from environmental water samples |
| url | https://doi.org/10.1111/1751-7915.14227 |
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