Rapid detection of methicillin-resistant staphylococci by multiplex PCR

A multiplex PCR was developed to detect the coagulase gene (coa; pathognomic of Staphylococcus aureus) and the mecA gene (characteristically encoding for methicillin resistance in staphylococci) in a single, rapid test. Suitable primers for the gene targets and an internal, amplification control were incorporated into a multiplex PCR assay, which was then optimized on a capillary air thermal cycler to improve the turnaround time of the test to approximately 1.5 hours. The assay was evaluated with 111 fresh clinical isolates of staphylococci. The multiplex PCR correctly distinguished between isolates of S. aureus, which were sensitive to methicillin (MSSA) and those resistant to it (MRSA). It also correctly differentiated between similar isolates of coagulase negative staphylococci (MSSE and MRSE respectively). It was concluded that this multiplex PCR was a rapid and reliable method for the detection of methicillin-resistant staphylococci.