Whole-Genome Multilocus Sequence Typing of Extended-Spectrum-Beta-Lactamase-Producing Enterobacteriaceae

Molecular typing has become indispensable in the detection of nosocomial transmission of bacterial pathogens and the identification of sources and routes of transmission in outbreak settings, but current methods are labor-intensive, are difficult to standardize, or have limited resolution. Whole-gen...

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Bibliographic Details
Published in:	Journal of clinical microbiology 2016-12, Vol.54 (12), p.2919-2927
Main Authors:	Kluytmans-van den Bergh, Marjolein F Q, Rossen, John W A, Bruijning-Verhagen, Patricia C J, Bonten, Marc J M, Friedrich, Alexander W, Vandenbroucke-Grauls, Christina M J E, Willems, Rob J L, Kluytmans, Jan A J W
Format:	Article
Language:	eng
Subjects:	Bacteriology beta-Lactamases - genetics beta-Lactamases - metabolism Citrobacter - classification Citrobacter - genetics Citrobacter - isolation & purification DNA, Bacterial - genetics Enterobacter cloacae - classification Enterobacter cloacae - genetics Enterobacter cloacae - isolation & purification Enterobacteriaceae - classification Enterobacteriaceae - genetics Enterobacteriaceae - isolation & purification Enterobacteriaceae Infections - microbiology Escherichia coli - classification Escherichia coli - genetics Escherichia coli - isolation & purification Humans Klebsiella oxytoca - classification Klebsiella oxytoca - genetics Klebsiella oxytoca - isolation & purification Klebsiella pneumoniae - classification Klebsiella pneumoniae - genetics Klebsiella pneumoniae - isolation & purification Multilocus Sequence Typing - methods
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Summary:	Molecular typing has become indispensable in the detection of nosocomial transmission of bacterial pathogens and the identification of sources and routes of transmission in outbreak settings, but current methods are labor-intensive, are difficult to standardize, or have limited resolution. Whole-genome multilocus sequence typing (wgMLST) has emerged as a whole-genome sequencing (WGS)-based gene-by-gene typing method that may overcome these limitations and has been applied successfully for several species in outbreak settings. In this study, genus-, genetic-complex-, and species-specific wgMLST schemes were developed for Citrobacter spp., the Enterobacter cloacae complex, Escherichia coli, Klebsiella oxytoca, and Klebsiella pneumoniae and used to type a national collection of 1,798 extended-spectrum-beta-lactamase-producing Enterobacteriaceae (ESBL-E) isolates obtained from patients in Dutch hospitals. Genus-, genetic-complex-, and species-specific thresholds for genetic distance that accurately distinguish between epidemiologically related and unrelated isolates were defined for Citrobacter spp., the E. cloacae complex, E. coli, and K. pneumoniae wgMLST was shown to have higher discriminatory power and typeability than in silico MLST. In conclusion, the wgMLST schemes developed in this study facilitate high-resolution WGS-based typing of the most prevalent ESBL-producing species in clinical practice and may contribute to further elucidation of the complex epidemiology of antimicrobial-resistant Enterobacteriaceae wgMLST opens up possibilities for the creation of a Web-accessible database for the global surveillance of ESBL-producing bacterial clones.
ISSN:	0095-1137 1098-660X