Department of Microbiology and Immunology, The University of Melbourne at the Doherty Institute for Infection & Immunity, Melbourne, Victoria, Australia.
Doherty Applied Microbial Genomics, The University of Melbourne at the Peter Doherty Institute for Infection & Immunity, Melbourne, Victoria, Australia.
Genome Med. 2018 Aug 31;10(1):63. doi: 10.1186/s13073-018-0572-z.
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 screening of mutations conferring pleiotropic phenotypes such as in vivo persistence, collateral sensitivity or cross-resistance. RM-seq will facilitate comprehensive detection, characterisation and surveillance of resistant bacterial populations ( https://github.com/rguerillot/RM-seq ).
突变获取是细菌抗生素耐药性的主要机制，但对其特征的描述仍不充分。在这里，我们介绍了 RM-seq，这是一种新的基于扩增子的高通量深度测序工作流程，基于一种从低错误扩增子测序 (LEA-seq) 改编而来的分子条形码技术。RM-seq 允许从混合细菌群体中高通量检测和功能评估突变耐药性。对非常低频耐药亚群的敏感检测允许对体外抗生素相关突变库进行特征描述，并在感染期间检测罕见的耐药群体。耐药突变的准确定量能够对赋予表型表型（如体内持久性、附带敏感性或交叉耐药性）的突变进行表型筛选。RM-seq 将有助于全面检测、描述和监测耐药细菌群体（https://github.com/rguerillot/RM-seq）。
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