Quantitative PCR versus metagenomics for monitoring antibiotic resistance genes: balancing high sensitivity and broad coverage.
gene quantification
limit of detection
limit of quantification
metagenomics sequencing
qPCR
sensitivity
Journal
FEMS microbes
ISSN: 2633-6685
Titre abrégé: FEMS Microbes
Pays: England
ID NLM: 9918227365806676
Informations de publication
Date de publication:
2023
2023
Historique:
received:
27
07
2022
revised:
24
02
2023
accepted:
05
03
2023
medline:
19
6
2023
pubmed:
19
6
2023
entrez:
19
6
2023
Statut:
epublish
Résumé
The widespread occurrence of clinically relevant antibiotic resistance within humans, animals, and environment motivates the development of sensitive and accurate detection and quantification methods. Metagenomics and quantitative PCR (qPCR) are amongst the most used approaches. In this study, we aimed to evaluate and compare the performance of these methods to screen antibiotic resistance genes in animal faecal, wastewater, and water samples. Water and wastewater samples were from hospital effluent, different treatment stages of two treatment plants, and of the receiving river at the discharge point. The animal samples were from pig and chicken faeces. Antibiotic resistance gene coverage, sensitivity, and usefulness of the quantitative information were analyzed and discussed. While both methods were able to distinguish the resistome profiles and detect gradient stepwise mixtures of pig and chicken faeces, qPCR presented higher sensitivity for the detection of a few antibiotic resistance genes in water/wastewater. In addition, the comparison of predicted and observed antibiotic resistance gene quantifications unveiled the higher accuracy of qPCR. Metagenomics analyses, while less sensitive, provided a markedly higher coverage of antibiotic resistance genes compared to qPCR. The complementarity of both methods and the importance of selecting the best method according to the study purpose are discussed.
Identifiants
pubmed: 37333442
doi: 10.1093/femsmc/xtad008
pii: xtad008
pmc: PMC10117749
doi:
Types de publication
Journal Article
Langues
eng
Pagination
xtad008Informations de copyright
© The Author(s) 2023. Published by Oxford University Press on behalf of FEMS.
Déclaration de conflit d'intérêts
None declared.
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