title: Development of a real-time PCR assay for the detection and identification of Staphylococcus capitis, Staphylococcus haemolyticus and Staphylococcus warneri.
abstract: Staphylococcus capitis, Staphylococcus haemolyticus and Staphylococcus warneri are coagulase-negative staphylococci. Each species has different characteristics, and a difference in pathology is also seen in compromised hosts. Therefore, the development of a species-specific simple detection method for the identification of these staphylococci is important. Here, a species-specific real-time PCR assay is reported that targets the superoxide dismutase A-encoding gene of these bacteria. Primers were designed with a base that was non-complementary with regard to the other bacteria. This base was at the 3' end of the primer (3' mismatch primer) and conferred high specificity. These primers were then evaluated using real-time PCR. They reacted only with the target bacterium. In addition, stable quantitative reactions were observed when experiments were performed using genomic DNA extracted from varying numbers of staphylococci cells (10(1)-10(7) cells). These results indicate that this method is useful for the identification and quantitative analysis of S. capitis, S. haemolyticus and S. warneri.
authors: Department of Microbiology II, Jikei University School of Medicine, 3-25-8 Nishi-Shinbashi, Minato-Ku, Tokyo 105-8461, Japan.
authors: Tadayuki
authors: T
authors: Iwase
authors: Department of Microbiology II, Jikei University School of Medicine, 3-25-8 Nishi-Shinbashi, Minato-Ku, Tokyo 105-8461, Japan.
authors: Keiko
authors: K
authors: Seki
authors: Department of Microbiology II, Jikei University School of Medicine, 3-25-8 Nishi-Shinbashi, Minato-Ku, Tokyo 105-8461, Japan.
authors: Hitomi
authors: H
authors: Shinji
authors: Department of Microbiology II, Jikei University School of Medicine, 3-25-8 Nishi-Shinbashi, Minato-Ku, Tokyo 105-8461, Japan.
authors: Yoshimitsu
authors: Y
authors: Mizunoe
authors: Department of Microbiology II, Jikei University School of Medicine, 3-25-8 Nishi-Shinbashi, Minato-Ku, Tokyo 105-8461, Japan.
authors: Shogo
authors: S
authors: Masuda
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D003030:Coagulase / Q000096:biosynthesis; D017931:DNA Primers / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013203:Staphylococcal Infections / Q000382:microbiology; D013210:Staphylococcus / Q000145:classification* / Q000302:isolation & purification*; D013482:Superoxide Dismutase / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D003030:Coagulase; D017931:DNA Primers; D013482:Superoxide Dismutase