Real-time PCR followed by high-resolution melting analysis – a new robust approach to evaluate SCCmec typing of methicillin-resistant Staphylococcus aureus

Abstract

Aim: We designed a novel approach based on real-time PCR followed by high-resolution melting (HRM) analysis to determine the Staphylococcal cassette chromosome mec (SCCmec) types of methicillin-resistant Staphylococcus aureus  strains, which we compared against the results of conventional multiplex PCR SCCmec typing. Methods: Multiplex PCR (for ccr and mec gene complexes) was carried out as conventional method. The HRM analysis was then designed using standard strains of each SCCmec type. Results: The M-PCR results included types III (33.33%), IV (43.33%) and V (23.33%). HRM analysis was able to distinguish all five types, which were used to set up the protocol with a sensitivity and specificity of 100% compared with the conventional method. Conclusion: This novel method for SCCmec typing has high specificity and sensitivity and can be conducted in a shorter period of time at lower costs.