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Sub-Typing of Extended-Spectrum-beta-Lactamase-Producing Isolates from a Nosocomial Outbreak: Application of a 10-Loci Generic Escherichia coli Multi-Locus Variable Number Tandem Repeat Analysis

Journal article
Authors Nahid Karami
Lisa Helldal
Christina Welinder-Olsson
Christina Åhrén
Edward R.B. Moore
Published in Plos One
Volume 8
Issue 12
Pages e83030
ISSN 1932-6203
Publication year 2013
Published at Institute of Biomedicine, Department of Infectious Medicine
Pages e83030
Language en
Links dx.doi.org/10.1371/journal.pone.008...
Keywords FIELD GEL-ELECTROPHORESIS, RAPID DETECTION, ENTEROBACTERIACEAE, STRAINS, CLONE, EPIDEMIOLOGY, COLLECTION, HOSPITALS, INFECTION
Subject categories Microbiology, Infectious Medicine

Abstract

Extended-spectrum beta-lactamase producing Escherichia coli (ESBL-E. coli) were isolated from infants hospitalized in a neonatal, post-surgery ward during a four-month-long nosocomial outbreak and six-month follow-up period. A multi-locus variable number tandem repeat analysis (MLVA), using 10 loci (GECM-10), for 'generic' (i.e., non-STEC) E. coli was applied for sub-species-level (i.e., sub-typing) delineation and characterization of the bacterial isolates. Ten distinct GECM-10 types were detected among 50 isolates, correlating with the types defined by pulsed-field gel electrophoresis (PFGE), which is recognized to be the 'gold-standard' method for clinical epidemiological analyses. Multi-locus sequence typing (MLST), multiplex PCR genotyping of bla(CTX-M), bla(TEM), bla(OXA) and bla(SHV) genes and antibiotic resistance profiling, as well as a PCR assay specific for detecting isolates of the pandemic O25b-ST131 strain, further characterized the outbreak isolates. Two clusters of isolates with distinct GECM-10 types (G06-04 and G07-02), corresponding to two major PFGE types and the MLST-based sequence types (STs) 131 and 1444, respectively, were confirmed to be responsible for the outbreak. The application of GECM-10 sub-typing provided reliable, rapid and cost-effective epidemiological characterizations of the ESBL-producing isolates from a nosocomial outbreak that correlated with and may be used to replace the laborious PFGE protocol for analyzing generic E. coli.

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