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Rapid identification of Streptococcus intermedius by PCR with the ily gene as a species marker gene

TAKATSUGU GOTO^*,, HIDEAKI NAGAMUNE^*, AIKO MIYAZAKI^*, YOSHIAKI KAWAMURA, OOKI OHNISHI^*, KANAKO HATTORI^*, KAZUTO OHKURA^*,||, KAZUAKI MIYAMOTO, SHIGERU AKIMOTO, TAKAYUKI EZAKI, KATSUHIKO HIROTA, YOICHIRO MIYAKE, TAKUYA MAEDA^* and HIROKI KOURAI^*

*Department of Biological Science and Technology, Faculty of Engineering, University of Tokushima, Minami-josanjima cho, Tokushima 770-8506, Department of Oral Microbiology, School of Dentistry, University of Tokushima, Kuramotocho, Tokushima 770-8504, Department of Microbiology, School of Medicine, Gifu University, Tsukasa-machi, Gifu 500-8705, Department of Microbiology, Wakayama Medical University, Kimiidera, Wakayama 641-0012 and ||Department of Applied Biological Sciences, Faculty of Agricultural Sciences, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan

Corresponding author: Dr H. Nagamune (e-mail: nagamune{at}bio.tokushima-u.ac.jp).

Received 10 April 2001; revised version accepted 16 August 2001.

Abstract

Streptococcus intermedius belongs to the anginosus group of streptococci (AGS) and is associated with endogenous infections leading to abscesses in the oral cavity and at deep-seated sites, such as the brain and liver. Two other species, S. anginosus and S. constellatus, and some presently unnamed taxa, are also classified as AGS. Recently, S. constellatus subsp. pharyngis, a new subspecies with biochemical characteristics similar to S. intermedius, was described with the potential for causing confusion when trying to identify isolates of these two species routinely with commercial identification kits, such as Rapid ID32 Strep and Fluo-Card Milleri. To correctly identify S. intermedius, this study attempted to develop an accurate PCR identification system with the ily gene as a species marker. This approach relies on amplification of an 819-bp fragment of the ily gene and its 3'-flanking region and is shown here to be specific for S. intermedius strains among all other streptococcal species. Moreover, this PCR system was applicable in direct rapid PCR with whole bacterial cells and TaKaRa Z-Taq^TM (TaKaRa), a highly efficient DNA polymerase, as the template and DNA amplification enzyme, respectively.

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