Abstract
Methicillin-resistant Staphylococcus aureus (MRSA) has become an important nosocomial pathogen, causing considerable morbidity and mortality. During the last 20 years, a variety of genotyping methods have been introduced for screening the prevalence of MRSA. In this study, we developed and evaluated an improved approach capillary gel electrophoresis based multilocus variable-number tandem-repeat fingerprinting (CGE/MLVF) for rapid MRSA typing. A total of 42 well-characterized strains and 116 non-repetitive clinical MRSA isolates collected from six hospitals in northeast China between 2009 and 2010 were tested. The results obtained by CGE/MLVF against clinical isolates were compared with traditional MLVF, spa typing, Multilocus sequence typing/staphylococcal cassette chromosome mec (MLST/SCCmec) and pulse field gel electrophoresis (PFGE). The discriminatory power estimated by Simpson’s index of diversity was 0.855 (28 types), 0.855 (28 patterns), 0.623 (11 types), 0.517 (8 types) and 0.854 (28 patterns) for CGE/MLVF, traditional MLVF, spa typing, MLST/SCCmec and PFGE, respectively. All methods tested showed a satisfied concordance in clonal complex level calculated by adjusted Rand’s coefficient. CGE/MLVF showed better reproducibility and accuracy than traditional MLVF and PFGE methods. In addition, the CGE/MLVF has potential to produce portable results. In conclusion, CGE/MLVF is a rapid and easy to use MRSA typing method with lower cost, good reproducibility and high discriminatory power for monitoring the outbreak and clonal spread of MRSA isolates.
Highlights
Methicillin-Resistant Staphylococcus aureus (MRSA) is a dangerous human pathogen, causing considerable morbidity and mortality worldwide
Traditional multilocus variable number tandem repeat fingerprinting (MLVF) was improved by combination multiplex PCR with capillary gel electrophoresis instead of traditional agar gel electrophoresis in order to obtain better discriminatory power and reproducibility
We demonstrated that Capillary Gel Electrophoresis (CGE)/MLVF was more suitable than traditional MLVF for typing Methicillin-resistant Staphylococcus aureus (MRSA)
Summary
Methicillin-Resistant Staphylococcus aureus (MRSA) is a dangerous human pathogen, causing considerable morbidity and mortality worldwide. Spa typing showed less discriminatory power than PFGE [11], its low cost, high reproducibility, appropriate stability, high throughput, and full data portability made this method the primary tool for characterization of MRSA isolates at the local and international level [3,12]. The discriminatory power of traditional MLVF was found to be comparable with that of PFGE and higher than that of MLST, other MLVA methods, spa typing and other PCR-based methods [11,14,15,16]. MRSA with newly designed set of primers for the same VNTR regions of traditional MLVF and combined it with micro-fluidic chip This new set of primers was considered to obtain more stable data. Criteria for clustering of CGE/MLVF patterns were proposed based on comparison with the data produced by spa typing, MLST and PFGE
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