In this study, a rapid and simple one-tube multiplex PCR (M-PCR) method was developed for Staphylococcal Cassette Chromosome mec (SCCmec), primary screening. One hundred and eight methicillin-resistant Staphylococcus aureus (MRSA) isolates with SCC mec typing results using the Oliveira and de Lencastre method (Oliveira and de Lencastre. Antimicrob. Agents Chemother. 46:2155-2161, 2002), were evaluated for the efficacy of our method. Typing results were also compared to the recently published PCR protocol by Kondo et al., (Kondo et al., Antimicrob Agents Chemother. 51:264-74, 2007). The one-tube M-PCR method showed comparable efficacy to the other two methods. None of the isolates were classified as non-typable in Kondo's and our method. Two formerly assigned SCCmec type I isolates by Oliveira and de Lencastre were determined as SCCmec type IV by Kondo's and the method in this study. In conclusion, the one-tube M-PCR method developed in this study increased the rapidity and simplicity for primary screening SCCmec type I to V MRSA.