Evaluation of high-resolution melting analysis for spa-typing of methicillin-resistant and -susceptible Staphylococcus aureus isolates.

Abstract

Molecular characterization of Staphylococcus aureus isolates from nosocomial and community-acquired infections using accurate, reproducible, and rapid typing methods is essential for the fast identification of prevalent and epidemic strains. Although sequence-based spa typing is highly effective, PCR-based techniques (such as high-resolution melting curve analysis, HRM) are simpler, less expensive, faster, and can be performed in a single and closed-tube assay format, thereby reducing the risk of contamination. A total of 51 methicillin-resistant S. aureus (MRSA) (n = 26) and methicillin-sensitive S. aureus (MSSA) (n = 25) isolates from Karaj (n = 10) and Yasuj (n = 41), Iran, were subjected to HRM. All selected isolates were identified by the standard spa-typing method. Among the 51 tested isolates, 11 genotype profiles were distinguished from 12 spa types. Strains t1077 and t1816 exhibited the highest and lowest melting temperatures (81.8°C and 79.4°C), with 46.7% and 39.8% G + C contents, respectively. Strains t706 and t1816, with almost identical G + C contents, had the same HRM genotypes, but their curves differed due to different G + C distributions. Four standard spa types (strains t030, t037, t701 and t5598) were differentiated correctly and their melting temperatures were 81.2°C, 81.4°C, 80.4°C and 80.1°C, respectively. We demonstrated that HRM profiling is a rapid method which enables the accurate screening of certain strains (especially the endemic ones), and may be used for bacterial surveillance. However, it cannot replace sequence-based spa typing, especially for newly emerging spa types, and therefore cannot be used as a standardized global method.