Development of rapid coagulase serotyping method by PCR and microplate hybridization.
We developed a novel PCR-based method for coagulase serotyping. Coagulase gene amplicons biotinylated by PCR were identified by microplate hybridization (MPH) using serotype-specific probes. The conventional serotyping method, which is strongly dependent on coagulase activity, may sometimes give a mistaken determination of the serotype, especially in cases where there is high coagulase activity. In contrast, the results of PCR-MPH are not affected by coagulase activity. Furthermore, once the isolated colonies are obtained, it only takes 3 h to perform PCR-MPH, and the interpretation of the results is entirely objective. We compared PCR-MPH with the conventional method for 90 strains of coagulase-producing Staphylococcus aureus. The same results were found for both the PCR-MPH and conventional methods, and thus, our results indicate that PCR-MPH is a rapid, objective, and reliable method for coagulase serotyping.
Kouguchi
Yoshihiro
Y
Fukuyama City Public Health Center, 2-11-22, Miyoshi-chou-minami, Fukuyama, Hiroshima 720-0032, Japan. shiken-kensa@city.fukuyama.hiroshima.jp
Fujiwara
Takako
T
Tamaoka
Yuko
Y
Teramoto
Miki
M
D015373:Bacterial Typing Techniques; D003030:Coagulase / Q000032:analysis* / Q000235:genetics; D017931:DNA Primers; D006225:Hand / Q000382:microbiology; D006801:Humans; D009693:Nucleic Acid Hybridization / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods*; D012703:Serotyping / Q000379:methods; D013211:Staphylococcus aureus / Q000145:classification* / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D013997:Time Factors
D003030:Coagulase; D017931:DNA Primers