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Journal of Clinical Microbiology, October 2008, p. 3246-3258, Vol. 46, No. 10
0095-1137/08/$08.00+0     doi:10.1128/JCM.00136-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Two Different Panton-Valentine Leukocidin Phage Lineages Predominate in Japan

Xiao Xue Ma,^1,7 Teruyo Ito,^1,2* Yoko Kondo,² Moe Cho,² Yukio Yoshizawa,³ Jun Kaneko,⁴ Atsuo Katai,⁵ Masato Higashiide,⁶ Shanshuang Li,² and Keiichi Hiramatsu^1,2

Department of Bacteriology, Juntendo University, Tokyo, Japan,¹ Department of Infection Control Science, Graduate School of Juntendo University, Tokyo, Japan,² Radioisotope Research Facility, The Jikei University School of Medicine, Tokyo, Japan,³ Laboratory of Applied Microbiology, Department of Microbial Biotechnology, Graduate School of Agricultural Science, Tohoku University, Sendai, Japan,⁴ Division of Laboratory Medicine, Kinan General Hospital, Wakayama, Japan,⁵ Kotobiken Medical Laboratories, Inc., Ibaragi, Japan,⁶ Department of Medical Microbiology and Parasitology, China Medical University, Shenyang, China⁷

Received 24 January 2008/ Returned for modification 27 April 2008/ Accepted 14 July 2008

We determined the entire nucleotide sequence of Sa2958-carrying Panton-Valentine leukocidin (PVL) gene, which was lysogenized in a sequence type 5 staphylococcal cassette chromosome mec (SCCmec) type II strain of methicillin-resistant Staphylococcus aureus (MRSA). Based on the nucleotide sequences of PVL phages, we developed PCRs to discriminate among five PVL phages, with a preliminary classification into two morphological groups (elongated-head type and icosahedral-head type) with four PCRs, including two PCRs for identifying the gene lineage between lukS-PV and the tail gene. The phages were then classified into five types by four PCRs identifying each phage-specific structure. With these PCRs, we examined the PVL phage types of 67 MRSA strains isolated in Japan from 1979 through 1985 and since 2000 and found that two morphologically distinct phages were predominant in Japan. The icosahedral-head-type phage, represented by the 108PVL type, was identified for 39 of 53 strains isolated from 1979 through 1985. Of 26 other Japanese isolates, 25 belonged either definitively or presumably to elongated-head types as follows: 3 belonged to the Sa2958 type; 8 were determined to belong to an elongated-head type, but a determination of greater specificity was not made; and 14 belonged to a Sa2958-like phage of unknown type. We induced prophages by treatment with mitomycin C from six strains of the Sa2958 type or of Sa2958-like unknown-type phages; five of six strains carried intact PVL-carrying phages, which can infect other S. aureus strains and might generate novel PVL-positive strains of S. aureus. That various SCCmec elements were carried by different strains of the same phage type suggests that S. aureus strains might independently acquire PVL phages before they acquire various SCCmec elements.

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* Corresponding author. Mailing address: Department of Bacteriology, Juntendo University, Hongo 2-1-1, Bunkyo-ku, Tokyo 113-8421, Japan. Phone: 011-3-5802-1041. Fax: 011-3-5684-7830. E-mail: teruybac{at}med.juntendo.ac.jp

Published ahead of print on 6 August 2008.

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Journal of Clinical Microbiology, October 2008, p. 3246-3258, Vol. 46, No. 10
0095-1137/08/$08.00+0     doi:10.1128/JCM.00136-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

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