Rapid detection of methicillin-resistant staphylococci by multiplex PCR

A multiplex PCR was developed to detect the coagulase gene (coa; pathognomi c of Staphylococcus aureus) and the mecA gene (characteristically encoding for methicillin resistance in staphylococci) in a single, rapid test. Suita ble primers for the gene targets and an internal, amplification control wer e incorporated into a multiplex PCR assay which was then optimized on a cap illary air thermal cycler to improve the turnaround time of the test to app roximately 1.5 hours. The assay was evaluated with 111 fresh clinical isola tes of staphylococci. The multiplex PCR correctly distinguished between iso lates of S. aureus, which were sensitive to methicillin (MSSA) and those re sistant to it (MRSA). It also correctly differentiated between similar isol ates of coagulase negative staphylococci (MSSE and MRSE respectively). It w as concluded that this multiplex PCR was a rapid and reliable method for th e detection of methicillin-resistant staphylococci. (C) 1999 The Hospital I nfection Society.