Use of Hybridization Probes in a Real-Time PCR Assay on the LightCycler for the Detection of Methicillin-Resistant Staphylococcus aureus

The rapid and accurate identification of methicillin-resistant Staphylococcus aureus (MRSA) is of great importance for the affected patient, the involved ward, and the microbiological laboratory. Resistance to methicillin is encoded by the mecA gene in S. aureus . Because routine laboratory diagnostics may be time consuming and because species differentiation encounters a variety of difficulties, molecular techniques detecting both the mecA and a S. aureus -specific gene are used for rapid and accurate detection and identification of MRSA. Various protocols, including the manual extraction of DNA have been established. In this chapter, the identification of MRSA based on simultaneous detection of the mecA gene and the S. aureus -specific Sa442 DNA fragment using automated DNA extraction and real-time polymerase chain reaction is described. This method is an attractive alternative to labor-intensive manual protocols and can easily be incorporated into the diagnostic microbiology laboratory workflow, with the ability to obtain results within 4 h.