Rapid identification of Streptococcus intermedius by PCR with the ily gene as a species marker gene.
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 deepseated 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 (TaKaRa), a highly efficient DNA polymerase, as the template and DNA amplification enzyme, respectively.
Goto
Takatsugu
T
*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.
Nagamune
Hideaki
H
*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.
Miyazaki
Aiko
A
*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.
Kawamura
Yoshiaki
Y
*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.
Ohnishi
Ooki
O
*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.
Hattori
Kanako
K
*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.
Ohkura
Kazuto
K
*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.
Miyamoto
Kazuaki
K
*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.
Akimoto
Shigeru
S
*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.
Ezaki
Takayuki
T
*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.
Hirota
Katsuhiko
K
*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.
Miyake
Yoichiro
Y
*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.
Maeda
Takuya
T
*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.
Kourai
Hiroki
H
*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.
D001426:Bacterial Proteins; D001430:Bacteriocins; D003603:Cytotoxins / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013291:Streptococcus / Q000302:isolation & purification*
D001426:Bacterial Proteins; D001430:Bacteriocins; D003603:Cytotoxins; C403910:intermedilysin protein, Streptococcus intermedius