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.