Identification of IncA/C plasmid replication and maintenance genes and development of a plasmid multilocus sequence typing scheme

Hancock, Steven J., Phan, Minh-Duy, Peters, Kate M., Forde, Brian M., Chong, Teik Min, Yin, Wai-Fong, Chan, Kok-Gan, Paterson, David L., Walsh, Timothy Rutland ORCID: https://orcid.org/0000-0003-4315-4096, Beatson, Scott A. and Schembri, Mark A. 2017. Identification of IncA/C plasmid replication and maintenance genes and development of a plasmid multilocus sequence typing scheme. Antimicrobial Agents and Chemotherapy 61 (2) , e01740-16. 10.1128/AAC.01740-16

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Abstract

Plasmids of incompatibility group A/C (IncA/C) are becoming increasingly prevalent within pathogenic Enterobacteriaceae. They are associated with the dissemination of multiple clinically relevant resistance genes, including blaCMY and blaNDM. Current typing methods for IncA/C plasmids offer limited resolution. In this study, we present the complete sequence of a blaNDM-1-positive IncA/C plasmid, pMS6198A, isolated from a multidrug-resistant uropathogenic Escherichia coli strain. Hypersaturated transposon mutagenesis, coupled with transposon-directed insertion site sequencing (TraDIS), was employed to identify conserved genetic elements required for replication and maintenance of pMS6198A. Our analysis of TraDIS data identified roles for the replicon, including repA, a toxin-antitoxin system; two putative partitioning genes, parAB; and a putative gene, 053. Construction of mini-IncA/C plasmids and examination of their stability within E. coli confirmed that the region encompassing 053 contributes to the stable maintenance of IncA/C plasmids. Subsequently, the four major maintenance genes (repA, parAB, and 053) were used to construct a new plasmid multilocus sequence typing (PMLST) scheme for IncA/C plasmids. Application of this scheme to a database of 82 IncA/C plasmids identified 11 unique sequence types (STs), with two dominant STs. The majority of blaNDM-positive plasmids examined (15/17; 88%) fall into ST1, suggesting acquisition and subsequent expansion of this blaNDM-containing plasmid lineage. The IncA/C PMLST scheme represents a standardized tool to identify, track, and analyze the dissemination of important IncA/C plasmid lineages, particularly in the context of epidemiological studies.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Medicine
Subjects:	R Medicine > R Medicine (General)
Uncontrolled Keywords:	Uropathogenic E. coli; IncA/C plasmid; Functional genomics; New Delhi metallo-beta-lactamase; Plasmid multilocus sequence typing
Publisher:	American Society for Microbiology
ISSN:	0066-4804
Date of First Compliant Deposit:	25 August 2017
Date of Acceptance:	8 November 2016
Last Modified:	07 May 2023 10:38
URI:	https://orca.cardiff.ac.uk/id/eprint/100805

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