Molecular Identification of CTX-M and BlaOXY/K1 Beta-lactamase Genes in Enterobacteriaceae by Sequencing of Universal M13-sequence Tagged PCR-amplicons

Overview

Journal BMC Infect Dis

Publisher Biomed Central

Specialty Infectious Diseases

Date 2009 Jan 24

PMID 19161622

Citations 11

Authors

Hans-Jurg Monstein

Maria Tarnberg

Lennart E Nilsson

Affiliations

Soon will be listed here.

Abstract

Background: Plasmid encoded blaCTX-M enzymes represent an important sub-group of class A beta-lactamases causing the ESBL phenotype which is increasingly found in Enterobacteriaceae including Klebsiella spp. Molecular typing of clinical ESBL-isolates has become more and more important for prevention of the dissemination of ESBL-producers among nosocomial environment.

Methods: Multiple displacement amplified DNA derived from 20 K. pneumoniae and 34 K. oxytoca clinical isolates with an ESBL-phenotype was used in a universal CTX-M PCR amplification assay. Identification and differentiation of blaCTX-M and blaOXY/K1 sequences was obtained by DNA sequencing of M13-sequence-tagged CTX-M PCR-amplicons using a M13-specific sequencing primer.

Results: Nine out of 20 K. pneumoniae clinical isolates had a blaCTX-M genotype. Interestingly, we found that the universal degenerated primers also amplified the chromosomally located K1-gene in all 34 K. oxytoca clinical isolates. Molecular identification and differentiation between blaCTX-M and blaOXY/K1-genes could only been achieved by sequencing of the PCR-amplicons. In silico analysis revealed that the universal degenerated CTX-M primer-pair used here might also amplify the chromosomally located blaOXY and K1-genes in Klebsiella spp. and K1-like genes in other Enterobacteriaceae.

Conclusion: The PCR-based molecular typing method described here enables a rapid and reliable molecular identification of blaCTX-M, and blaOXY/K1-genes. The principles used in this study could also be applied to any situation in which antimicrobial resistance genes would need to be sequenced.

Citing Articles

Evaluation of Phenotypic Tests to Detect Extended-Spectrum β-Lactamase (ESBL)-Producing Klebsiella oxytoca Complex Strains.

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ROCker Models for Reliable Detection and Typing of Short-Read Sequences Carrying β-Lactamase Genes.

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Duration of travel-associated faecal colonisation with ESBL-producing Enterobacteriaceae - A one year follow-up study.

OstholmBalkhed A, Tarnberg M, Nilsson M, Nilsson L, Hanberger H, Hallgren A PLoS One. 2018; 13(10):e0205504.

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Survey of Gene Frequency in Extended-Spectrum Beta-Lactamase-Producing Isolates Using the Combination Disk and PCR Methods in Ahvaz, Iran.

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