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Performance characteristics of next-generation sequencing for antimicrobial resistance gene detection in genomes and metagenomes

Ashley M. Rooney, Amogelang R. Raphenya, Roberto G. Melano, Christine Seah, Noelle R. Yee, Derek R. MacFadden, Andrew G. McArthur, Pierre H.H. Schneeberger, Bryan Coburn
doi: https://doi.org/10.1101/2021.06.25.449921
Ashley M. Rooney
aDepartment of Laboratory Medicine and Pathobiology, Faculty of Medicine, University of Toronto, Toronto, ON, Canada
bUniversity Health Network, Division of Infectious Diseases and Toronto General Hospital Research Institute, Toronto, ON, Canada
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Amogelang R. Raphenya
cDavid Braley Centre for Antibiotic Discovery, McMaster University, Hamilton, ON, Canada
dMichael G. DeGroote Institute for Infectious Disease Research, McMaster University, Hamilton, ON, Canada
eDepartment of Biochemistry and Biomedical Science, McMaster University, Hamilton, ON, Canada
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Roberto G. Melano
aDepartment of Laboratory Medicine and Pathobiology, Faculty of Medicine, University of Toronto, Toronto, ON, Canada
fPublic Health Ontario Laboratory, Toronto, ON, Canada
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Christine Seah
fPublic Health Ontario Laboratory, Toronto, ON, Canada
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Noelle R. Yee
bUniversity Health Network, Division of Infectious Diseases and Toronto General Hospital Research Institute, Toronto, ON, Canada
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Derek R. MacFadden
gOttawa Hospital Research Institute, Ottawa, ON, Canada
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Andrew G. McArthur
cDavid Braley Centre for Antibiotic Discovery, McMaster University, Hamilton, ON, Canada
dMichael G. DeGroote Institute for Infectious Disease Research, McMaster University, Hamilton, ON, Canada
eDepartment of Biochemistry and Biomedical Science, McMaster University, Hamilton, ON, Canada
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Pierre H.H. Schneeberger
aDepartment of Laboratory Medicine and Pathobiology, Faculty of Medicine, University of Toronto, Toronto, ON, Canada
bUniversity Health Network, Division of Infectious Diseases and Toronto General Hospital Research Institute, Toronto, ON, Canada
hDepartment of Medical Parasitology and Infection Biology, Swiss Tropical and Public Health Institute, University of Basel, Basel, Switzerland
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  • For correspondence: pierre.schneeberger@swisstph.ch
Bryan Coburn
aDepartment of Laboratory Medicine and Pathobiology, Faculty of Medicine, University of Toronto, Toronto, ON, Canada
bUniversity Health Network, Division of Infectious Diseases and Toronto General Hospital Research Institute, Toronto, ON, Canada
iDepartment of Medicine, Faculty of Medicine, University of Toronto, Toronto, ON, Canada
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  • For correspondence: bryan.coburn@uhn.ca
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Abstract

Short-read sequencing provides a culture-independent method for the detection of antimicrobial resistance (AMR) genes from single bacterial genomes and metagenomic samples. However, the performance characteristics of these approaches have not been systematically characterized. We compared assembly- and read-based approaches to determine sensitivity, positive predictive value, and sequencing limits of detection required for AMR gene detection in an Escherichia coli ST38 isolate spiked into a synthetic microbial community at varying abundances. Using an assembly-based method the limit of detection was 15X genome coverage. We are confident in AMR gene detection at target relative abundances of 100% to 1%, where a target abundance of 1% would require assembly of approximately 30 million reads to achieve 15X target coverage. Recent studies assessing AMR gene content in metagenomic samples may be inadequately sequenced to achieve high sensitivity. Our study informs future sequencing projects and analytical strategies for genomic and metagenomic AMR gene detection.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted June 26, 2021.
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Performance characteristics of next-generation sequencing for antimicrobial resistance gene detection in genomes and metagenomes
Ashley M. Rooney, Amogelang R. Raphenya, Roberto G. Melano, Christine Seah, Noelle R. Yee, Derek R. MacFadden, Andrew G. McArthur, Pierre H.H. Schneeberger, Bryan Coburn
bioRxiv 2021.06.25.449921; doi: https://doi.org/10.1101/2021.06.25.449921
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Performance characteristics of next-generation sequencing for antimicrobial resistance gene detection in genomes and metagenomes
Ashley M. Rooney, Amogelang R. Raphenya, Roberto G. Melano, Christine Seah, Noelle R. Yee, Derek R. MacFadden, Andrew G. McArthur, Pierre H.H. Schneeberger, Bryan Coburn
bioRxiv 2021.06.25.449921; doi: https://doi.org/10.1101/2021.06.25.449921

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