To confirm the presence of other pathogens in the Kagoshima sample, detection of agents causing diarrhea using our real-time PCR system, referred to as “Dembo-PCR,” was performed . This system can identify 19 species of pathogens, including virus, bacteria and protozoa. Briefly, viral DNA and RNA were extracted by high pure viral nucleic acid extraction kit (Roche Diagnostics GmbH, Mannheim, Germany) and bacteria and protozoa DNA were extracted by QIAamp Fast DNA stool mini kit (QIAGEN, Hilden, Germany). Nucleic acids extracted by each kit were subjected to Dembo-PCR, according to a previous report .