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Rapid molecular identification of imported mycoses in Japan

日本输入性真菌病的快速分子鉴定

基本信息

批准号：
11670259
负责人：
MIKAMI Yuzuru
金额：
$ 1.73万
依托单位：
Chiba University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670259/
关键词：
Imported mycoses gene analysis PCR primer genotypes ITS region 病原真菌遺伝子 PCRプライマー Paracoccidioides Cryptococcus

项目摘要

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.

随着日本国际化程度的大幅提高，不少日本人有机会出国旅游或工作。然而，经常有报道称，其中一些国家患有东亚（泰国、中国、印度尼西亚）、美国、巴西等国家特有的严重真菌流行病。这些感染被称为输入性真菌病：球孢子菌病、组织胞浆菌病、马梅菲青霉病和副球孢子菌病。由于这些病原体处理起来非常危险（第 3 类），因此临床实验室需要简单、安全的识别和诊断系统。本项目的目的是为这些输入性真菌感染的病原体（H.casulatum、P.mameffei、P.brasiliensis 和 C.neoformans）制备简单、安全的分子鉴定系统。 1）使用（ITS）区域（ITS1-5.8S rDNA-ITS2）序列分析对代表 var.capsulatum、duboisii 和farciminosum 三个品种的 29 个地理上不同的 H.capsulatum 分离株的系统发育进行评估，我们发现 ITS 区域包含足够的信息来解析真菌分离株之间的系统发育关系。 2）根据ITS区序列信息，制备新的PCR引物（设计扩增418 bp），用于巴西P.brasiliensis的分子鉴定。 3) ITS 区域分析使我们能够将新隐球菌 var.gattii 的所有分离株区分为四个地理组。 4) P.mameffei（由我们研究组开发）的PCR引物对的有用性也得到了证实。因此，针对日本输入性真菌病的每种病原体新制备的PCR引物对很简单，并且被发现可用于临床实验室的分子鉴定。