Rapid and robust phylotyping of spa t003, a dominant MRSA clone in Luxembourg and other European countries

BMC Infectious Diseases

Jos Even¹  Paul Keim²  Joel Mossong¹  James M Schupp³  Frederic Decruyenaere¹  Jesse Trujillo³  Carl F Eberhard³  Jolene Bowers³  Elizabeth M Driebe³  Erin Kelley³  David M Engelthaler³ 

[1] Laboratoire National de Santé, Luxembourg, USA;Northern Arizona University, Flagstaff, AZ, USA;Translational Genomics Research Institute, 3051 W. Shamrell Blvd, 86001, Flagstaff, AZ, USA

关键词: Molecular epidemiology;    Phylotyping;    Luxembourg;    WGST;    SNP assays;    MRSA;    spa t003;   

Others  :  1146789
DOI  :  10.1186/1471-2334-13-339

 received in 2013-01-12, accepted in 2013-07-18,  发布年份 2013

【 摘 要 】

Background

spa typing is a common genotyping tool for methicillin-resistant Staphylococcus aureus (MRSA) in Europe. Given the high prevalence of dominant clones, spa-typing is proving to be limited in its ability to distinguish outbreak isolates from background isolates. New molecular tools need to be employed to improve subtyping of dominant local MRSA strains (e.g., spa type t003).

Methods

Phylogenetically critical, or canonical, SNPs (can-SNPs) were identified as subtyping targets through sequence analysis of 40 MRSA whole genomes from Luxembourg. Real-time PCR assays were designed around target SNPs and validated using a repository of 240 previously sub-typed and epidemiologically characterized Luxembourg MRSA isolates, including 153 community and hospital isolates, 69 isolates from long term care (LTC) facilities, and 21 prospectively analyzed MRSA isolates. Selected isolates were also analyzed by whole genome SNP typing (WGST) for comparison to the SNP assays and other subtyping techniques.

Results

Fourteen real-time PCR assays were developed and validated, including two assays to determine presence of spa t003 or t008. The other twelve assays successfully provided a high degree of resolution within the t003 subtype. WGST analysis of the LTC facility isolates provided greater resolution than other subtyping tools, identifying clusters indicative of ongoing transmission within LTC facilities.

Conclusions

canSNP-based PCR assays are useful for local level MRSA phylotyping, especially in the presence of one or more dominant clones. The assays designed here can be easily adapted for investigating t003 MRSA strains in other regions in Western Europe. WGST provides substantially better resolution than other typing methods.

【 授权许可】

   
2013 Engelthaler et al.; licensee BioMed Central Ltd.

【 图 表 】

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