Rapid molecular identification of imported mycoses in Japan

点击翻译按钮获取中文摘要

Coupled with the large increase in Japanese internationalization, a number of Japanese people have had opportunities to go abroad for oversea travel or work purposes. However, frequently it has been reported that some of them had suffered from serious fungal endemics peculiar to the countries such as East Asia (Thailand, China, Indonesia), USA, Brazil and others. These infections are called as imported mycoses : coccidioidomycosis, histoplasmosis, penicilliosis mameffei and paracoccidioidomycosis. Since these causative agents are very dangerous to handle (class 3), simple and safe identification and diagnostic systems are requested in clinical laboratory.In this project, preparation of simple and safty molecular identification system for the etiological agents of these imported fungal infection (H.casulatum, P.mameffei, P.brasiliensis and C.neoformans) was aimed. 1) Phylogeny of twenty nine geographically diverse H.capsulatum isolates representing the three varieties, var.capsulatum, duboisii and farciminosum were evaluated using (ITS) region (ITS1-5.8S rDNA-ITS2) sequence analyses, and we found that the ITS region contained sufficient information to resolve the phylogenetic relationship among the fungal isolates. 2) Based on the ITS region sequence information, a new PCR primer (designed to amplify 418 bp) was prepared for the molecular identification of P.brasiliensis. 3) ITS region analysis of allowed us to distinguish all isolates of C.neoformans var.gattii into four geographic groups. 4) Usefulness of PCR primer pairs for P.mameffei (developed by our research group) was also confirmed.Thus newly prepared PCR primer pairs for each etiological agent of imported mycoses in Japan were simple and found to be useful for the molecular identification in clinical laboratory.