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Strategy for rapid detection of carbapenemase-producing Enterobacteriaceae.

ABSTRACT: A prospective survey was conducted on 862 Enterobacteriaceae isolates with reduced susceptibility to carbapenems. The Carba NP test, UV spectrophotometry, and a DNA microarray were used to detect carbapenemase producers, and the results were compared to those from PCR and sequencing. The 172 carbapenemase producers were detected using the Carba NP test and UV spectrophotometry, whereas the DNA microarray failed to detect IMI producers. The use of the Carba NP test as a first screening, followed by the use of molecular techniques, has been determined to be an efficient strategy for identifying carbapenemase-producing Enterobacteriaceae.

SUBMITTER: Dortet L

PROVIDER: S-EPMC4023735 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature

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Strategy for rapid detection of carbapenemase-producing Enterobacteriaceae.

Dortet Laurent L Bréchard Ludivine L Cuzon Gaëlle G Poirel Laurent L Nordmann Patrice P

Antimicrobial agents and chemotherapy 20140127 4

A prospective survey was conducted on 862 Enterobacteriaceae isolates with reduced susceptibility to carbapenems. The Carba NP test, UV spectrophotometry, and a DNA microarray were used to detect carbapenemase producers, and the results were compared to those from PCR and sequencing. The 172 carbapenemase producers were detected using the Carba NP test and UV spectrophotometry, whereas the DNA microarray failed to detect IMI producers. The use of the Carba NP test as a first screening, followed ...[more]

PMID: 24468779

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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

Strategy for rapid detection of carbapenemase-producing Enterobacteriaceae.

Publications

Strategy for rapid detection of carbapenemase-producing Enterobacteriaceae.

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