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Genomic analysis of carbapenemase-producing Enterobacteriaceae in Queensland reveals widespread transmission of bla _IMP-4 on an IncHI2 plasmid.

ABSTRACT: Carbapenemase-producing Enterobacteriaceae (CPE) are an increasingly common cause of healthcare-associated infections and may occasionally be identified in patients without extensive healthcare exposure. bla _IMP-4 is the most frequently detected carbapenemase gene in Enterobacteriaceae within Australia, but little is known about the mechanisms behind its persistence. Here we used whole genome sequencing (WGS) to investigate the molecular epidemiology of bla _IMP-4 in Queensland, Australia. In total, 107 CPE were collected between 2014 and 2017 and sent for WGS on an Illumina NextSeq500. Resistance genes and plasmid types were detected using a combination of read mapping and nucleotide comparison of de novo assemblies. Six isolates were additionally sequenced using Oxford Nanopore MinION to generate long-reads and fully characterize the context of the bla _IMP-4 gene. Of 107 CPE, 93 carried the bla _IMP-4 gene; 74/107 also carried an IncHI2 plasmid, suggesting carriage of the bla _IMP-4 gene on an IncHI2 plasmid. Comparison of these isolates to a previously characterized IncHI2 plasmid pMS7884A (isolated from an Enterobacter hormaechei strain in Brisbane) suggested that all isolates carried a similar plasmid. Five of six representative isolates sequenced using Nanopore long-read technology carried IncHI2 plasmids harbouring the bla _IMP-4 gene. While the vast majority of isolates represented ?E. hormaechei, several other species were also found to carry the IncHI2 plasmid, including Klebsiella species, Escherichia coli and Citrobacter species. Several clonal groups of E. hormaechei were also identified, suggesting that persistence of bla _IMP-4 is driven by both presence on a common plasmid and clonal spread of certain E. hormaechei lineages.

SUBMITTER: Roberts LW

PROVIDER: S-EPMC7067041 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature

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Genomic analysis of carbapenemase-producing Enterobacteriaceae in Queensland reveals widespread transmission of blaIMP-4 on an IncHI2 plasmid.

Roberts Leah W LW Catchpoole Elizabeth E Jennison Amy V AV Bergh Haakon H Hume Anna A Heney Claire C George Narelle N Paterson David L DL Schembri Mark A MA Beatson Scott A SA Harris Patrick N A PNA

Microbial genomics 20191219 1

Carbapenemase-producing Enterobacteriaceae (CPE) are an increasingly common cause of healthcare-associated infections and may occasionally be identified in patients without extensive healthcare exposure. blaIMP-4 is the most frequently detected carbapenemase gene in Enterobacteriaceae within Australia, but little is known about the mechanisms behind its persistence. Here we used whole genome sequencing (WGS) to investigate the molecular epidemiology of bla< ...[more]

PMID: 31860437

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Project description:BackgroundCarbapenem-resistant Enterobacteriaceae (CRE) are responsible for worldwide outbreaks and antibiotic treatments are problematic. The polysaccharide poly-(?-1,6)-N-acetyl glucosamine (PNAG) is a vaccine target detected on the surface of numerous pathogenic bacteria, including Escherichia coli. Genes encoding PNAG biosynthetic proteins have been identified in two other main pathogenic Enterobacteriaceae, Enterobacter cloacae and Klebsiella pneumoniae. We hypothesized that antibodies to PNAG might be a new therapeutic option for the different pan-resistant pathogenic species of CRE.MethodsPNAG production was detected by confocal microscopy and its role in the formation of the biofilm (for E. cloacae) and as a virulence factor (for K. pneumoniae) was analysed. The in vitro (opsonophagocytosis killing assay) and in vivo (mouse models of peritonitis) activity of antibodies to PNAG were studied using antibiotic-susceptible and -resistant E. coli, E. cloacae and K. pneumoniae. A PNAG-producing strain of Pseudomonas aeruginosa, an organism that does not naturally produce this antigen, was constructed by adding the pga locus to a strain with inactive alg genes responsible for the production of P. aeruginosa alginate. Antibodies to PNAG were tested in vitro and in vivo as above.ResultsPNAG is a major component of the E. cloacae biofilm and a virulence factor for K. pneumoniae. Antibodies to PNAG mediated in vitro killing (>50%) and significantly protected mice against the New Delhi metallo-?-lactamase-producing E. coli (P?=?0.02), E. cloacae (P?=?0.0196) and K. pneumoniae (P?=?0.006), against K. pneumoniae carbapenemase (KPC)-producing K. pneumoniae (P?=?0.02) and against PNAG-producing P. aeruginosa (P?=?0.0013). Thus, regardless of the Gram-negative bacterial species, PNAG expression is the sole determinant of the protective efficacy of antibodies to this antigen.ConclusionsOur findings suggest antibodies to PNAG may provide extended-spectrum antibacterial protective activity.

Dataset Information

Genomic analysis of carbapenemase-producing Enterobacteriaceae in Queensland reveals widespread transmission of bla _IMP-4 on an IncHI2 plasmid.

Publications

Genomic analysis of carbapenemase-producing <i>Enterobacteriaceae</i> in Queensland reveals widespread transmission of <i>bla</i><sub>IMP-4</sub> on an IncHI2 plasmid.

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Dataset Information

Genomic analysis of carbapenemase-producing Enterobacteriaceae in Queensland reveals widespread transmission of bla IMP-4 on an IncHI2 plasmid.

Publications

Genomic analysis of carbapenemase-producing <i>Enterobacteriaceae</i> in Queensland reveals widespread transmission of <i>bla</i><sub>IMP-4</sub> on an IncHI2 plasmid.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Genomic analysis of carbapenemase-producing Enterobacteriaceae in Queensland reveals widespread transmission of bla _IMP-4 on an IncHI2 plasmid.