Journal article

Comprehensive antibiotic-linked mutation assessment by resistance mutation sequencing (RM-seq)

Romain Guerillot, Lucy Li, Sarah Baines, Brian Howden, Mark B Schultz, Torsten Seemann, Ian Monk, Sacha J Pidot, Wei Gao, Stefano Giulieri, Anders Goncalves da Silva, Anthony D'Agata, Takehiro Tomita, Anton Y Peleg, Timothy P Stinear, Benjamin P Howden

GENOME MEDICINE | BMC | Published : 2018

Abstract

Mutation acquisition is a major mechanism of bacterial antibiotic resistance that remains insufficiently characterised. Here we present RM-seq, a new amplicon-based deep sequencing workflow based on a molecular barcoding technique adapted from Low Error Amplicon sequencing (LEA-seq). RM-seq allows detection and functional assessment of mutational resistance at high throughput from mixed bacterial populations. The sensitive detection of very low-frequency resistant sub-populations permits characterisation of antibiotic-linked mutational repertoires in vitro and detection of rare resistant populations during infections. Accurate quantification of resistance mutations enables phenotypic screeni..

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Grants

Awarded by National Health and Medical Research Council (NHMRC), Australia


Funding Acknowledgements

This work was supported by the National Health and Medical Research Council (NHMRC), Australia project grant (GNT1066791) and Research Fellowship to TPS (GNT1008549) and Practitioner Fellowship to BPH (GNT1105905). Doherty Applied Microbial Genomics is funded by the Department of Microbiology and Immunology at The University of Melbourne.