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Research Article

Real-time PCR Followed by High-Resolution Melting Analysis – a New Robust Approach to Evaluate SCCmec Typing of Methicillin-Resistant Staphylococcus Aureus

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Pages 155-164 | Received 26 Jun 2018, Accepted 26 Nov 2018, Published online: 22 Jan 2019
 

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 aureusstrains, 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.

Financial & competing interests disclosure

This research was based on an MSc thesis supported by Tehran University of Medical Sciences (grant no. 35167). The authors have no other financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Additional information

Funding

This research was based on an MSc thesis supported by Tehran University of Medical Sciences (grant no. 35167). The authors have no other financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed

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