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Journal of Clinical Microbiology, November 2004, p. 4925-4930, Vol. 42, No. 11
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.11.4925-4930.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Development of a PCR Method for Rapid Identification of New Streptococcus mutans Serotype k Strains

Kazuhiko Nakano,1 Ryota Nomura,1 Noriko Shimizu,1 Ichiro Nakagawa,2 Shigeyuki Hamada,2 and Takashi Ooshima1*

Departments of Pediatric Dentistry,1 Oral and Molecular Microbiology, Osaka University Graduate School of Dentistry, Suita, Osaka, Japan2

Received 19 March 2004/ Returned for modification 28 June 2004/ Accepted 6 July 2004

In a previous study, we isolated and characterized a new serotype k of Streptococcus mutans from human blood and oral cavities. Analysis of the genes involved in biosynthesis of the serotype-specific polysaccharide of serotype k strains revealed that the serotype k-specific nucleotide alignment was commonly present in the 5' region of the rgpF gene (350 bp from the initial sequence) compared to the reference strains, and then a method for rapid identification of serotype k strains was developed by use of PCR with primers designed on the basis of the sequence of the variable region. PCR assays with primers specific for amplification of serotype k strains showed a negative reaction with serotype c, e, and f strains and a positive reaction with serotype k strains, with the sensitivity for identification of the serotype k strains shown to range from 5 to 50 cells. Next, the frequency of positive reactions for serotype k-specific primers was surveyed with DNA taken from saliva samples from 200 subjects (2 to 18 years of age), and 10 of those showed a positive reaction, which was higher than the frequency in our previous survey with a serological method. In addition, all saliva samples from subjects with serotype k strains in our previous study were shown to be positive with the serotype k-specific primers. These results indicate that this new PCR method is effective for identification of subjects with S. mutans serotype k.

* Corresponding author. Mailing address: Department of Pediatric Dentistry, Osaka University Graduate School of Dentistry, 1-8 Yamada-oka, Suita, Osaka 565-0871, Japan. Phone: 81 6 6879 2961. Fax: 81 6 6879 2965. E-mail: ooshima{at}dent.osaka-u.ac.jp.

Journal of Clinical Microbiology, November 2004, p. 4925-4930, Vol. 42, No. 11
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.11.4925-4930.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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