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Antimicrobial Agents and Chemotherapy, January 2007, p. 264-274, Vol. 51, No. 1
0066-4804/07/$08.00+0     doi:10.1128/AAC.00165-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Combination of Multiplex PCRs for Staphylococcal Cassette Chromosome mec Type Assignment: Rapid Identification System for mec, ccr, and Major Differences in Junkyard Regions{triangledown}

Yoko Kondo,1 Teruyo Ito,1,2* Xiao Xue Ma,2 Shinya Watanabe,1 Barry N. Kreiswirth,3 Jerome Etienne,4 and Keiichi Hiramatsu1,2

Juntendo University, Graduate School of Medicine, Department of Infection Control Science, Tokyo, Japan,1 Juntendo University, Department of Bacteriology, Tokyo, Japan,2 Public Health Research Institute, Newark, New Jersey,3 Faculté de Médecine Laennec, Centre National de Référence des Staphylocoques, IFR62, INSERM E02030, 7 rue guillaume Paradin, 69008 Lyon, France4

Received 8 February 2006/ Returned for modification 7 April 2006/ Accepted 9 October 2006

Staphylococcal cassette chromosome mec (SCCmec) typing, in combination with genotyping of the Staphylococcus aureus chromosome, has become essential for defining methicillin-resistant S. aureus (MRSA) clones in epidemiological studies. We have developed a convenient system for SCCmec type assignment. The system consists of six multiplex PCRs (M-PCRs) for identifying the ccr gene complex (ccr), the mec gene complex (mec), and specific structures in the junkyard (J) regions: M-PCR with primer set 1 (M-PCR 1) identified five types of ccr genes; M-PCR 2 identified class A to class C mec; M-PCRs 3 and 4 identified specific open reading frames in the J1 regions of type I and IV and of type II, III, and V SCCmec elements, respectively; M-PCR 5 identified the transposons Tn554 and {Psi}Tn554 integrated into the J2 regions of type II and III SCCmec elements; and M-PCR 6 identified plasmids pT181 and pUB110 integrated into J3 regions. The system was validated with 99 MRSA strains carrying SCCmec elements of different types. The SCCmec types of 93 out of the 99 MRSA strains could be assigned. The SCCmec type assignments were identical to those made with a PCR system that uses numerous primer pairs to identify genes or gene alleles. Our system of six M-PCRs is thus a convenient and reliable method for typing SCCmec elements.

* Correspondingauthor. Mailing address: Department of Bacteriology, Juntendo University, 2-1-1 Hongo, Bunkyo-ku, Tokyo 113-8421, Japan. Phone: 81-3-5802-1041. Fax: 81-3-5684-7830. E-mail: teruybac{at}med.juntendo.ac.jp.

{triangledown} Published ahead of print on 16 October 2006.

Antimicrobial Agents and Chemotherapy, January 2007, p. 264-274, Vol. 51, No. 1
0066-4804/07/$08.00+0     doi:10.1128/AAC.00165-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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