Rapid detection of hyper virulent strains of Clostridium difficile and application to direct detection from stool specimens

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A typing system for Clostridium difficile by sequencing the surface layer protein A gene (slpA) was evaluated, and clinical isolates in Japan were analyzed by this method. slpA sequence typing was found to have reliable typability and discriminatory power, and the typing results were highly reproducible and comparable. slpA sequence typing could be applied to direct typing successfully. The emerging strain, PCR ribotype 027 was recovered from one of the 160 specimens tested. Based on the sequence data of slpA, loop-mediated isothermal amplification (LAMP) method detecting slpA of PCR ribotype 027 (slpAgc8) was established. The LAMP assay detecting slpAgc8 appears to be a valuable tool for the rapid identification of this type.