Rapid culture-independent loop-mediated isothermal amplification detection of antimicrobial resistance markers from environmental water samples.
Microb Biotechnol
; 16(5): 977-989, 2023 05.
Article
in English
| MEDLINE | ID: covidwho-2296253
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.
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Main subject:
Escherichia coli Proteins
/
Anti-Bacterial Agents
Type of study:
Diagnostic study
/
Observational study
Limits:
Animals
/
Humans
Language:
English
Journal:
Microb Biotechnol
Year:
2023
Document Type:
Article
Affiliation country:
1751-7915.14227
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