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Methicillin-resistant Staphylococcus aureus (MRSA) continues to be a burden to the global healthcare system. Locally, the prevalence of MRSA in New Zealand has increased significantly over the last 10 years underscoring the urgency to control the colonisation and spread of MRSA. Following on from a previous multi-centre study looking at MRSA colonisation in the nares and throats of a New Zealand population using a PCR method, this study aims to determine the usefulness of DNA melting curve analysis in the differentiation of MRSA strains. Specifically investigated is a possible relationship between the isolate DNA melting temperature (TM) and MRSA strain. Bacterial typing can be used to confirm or refute the relatedness of isolates and to plan MRSA control programmes and while the determination of MRSA strains is currently done using staphylococcal protein A gene typing based upon repeat pattern analysis and pulse-field gel electrophoresis, these processes are often time consuming, require specialist skills, and are performed at reference laboratories. In this study, 10 MRSA isolates where detected from a total of 270 volunteers using the Roche LightCycler MRSA Advanced Test. It was found that the WSPP was the most frequently isolated strain. In addition, by using the DNA melting temperatures, an apparent link was found: the WSPP strain had the highest TM, followed by the AK3 strain, and lastly, the WK/AK1 strain had the lowest TM. Moreover, no variation in TM was observed between different volunteer isolates of the same strain type (WSPP, AK3, and WK/AK1 displayed consistent TM values of 59.6 o C, 58.1 o C, and 57.7 o C, respectively). The significance of swiftly determining the MRSA strain is somewhat understated. From an infection control/epidemiology perspective, it provides information on origin and mutation shift of the pathogen, helping in the control of MRSA epidemics, thus contributing to our knowledge of strain dynamics. More importantly perhaps, it provides information on determining the course of treatment for the patient as strains differ in their antimicrobial susceptibility patterns. While it is too early to draw conclusions from this data, it certainly warrants further investigation.

iosr journal of dental and medical sciences

Detection and simultaneous confirmation of Methicillin-Resistant Staphylococcus aureus by an automated Real-Time PCR method with strain differentiation by analysis of DNA melting curve characteristics