新型隐球菌格鲁比变种的多位点微卫星分型

Multilocus microsatellite typing for Cryptococcus neoformans var. grubii.

作者信息

Hanafy Ahmed, Kaocharoen Sirada, Jover-Botella Alejandro, Katsu Masakazu, Iida Soji, Kogure Takahisa, Gonoi Tohru, Mikami Yuzuru, Meyer Wieland

机构信息

Medical Mycology Research Center (former Research Center for Pathogenic Fungi and Microbial Toxicoses), Chiba University, Chiba, Japan.

出版信息

Med Mycol. 2008 Nov;46(7):685-96. doi: 10.1080/13693780802027062.

DOI:10.1080/13693780802027062

PMID:18608927

Abstract

Fifteen randomly selected microsatellites (simple sequence repeats; SSRs), from the H99 Cryptococcus neoformans var. grubii (serotype A) genome, were sequenced, characterized and applied to sequence 87 clinical and environmental C. neoformans var. grubii isolates from 12 different countries based on Multilocus Microsatellite Typing (MLMT). Among the 15 SSR loci, three (designated CNG1, CNG2 and CNG3) were polymorphic, while the remaining 12 SSR loci showed no variations. The specific PCR primers of the polymorphic microsatellites, i.e., CNG1, CNG2 and CNG3, amplified those loci only from strains of C. neoformans (C. neoformans var. grubii, C. neoformans var. neoformans and the AD hybrid) but not from Cryptococcus gattii, suggesting a species-specific association. The three polymorphic microsatellites are useful markers for strain genotyping, population genetic analysis, epidemiological studies, and may be helpful for the diagnosis of cryptococcosis due to C. neoformans.

摘要

从新型隐球菌格鲁比变种（血清型A）H99基因组中随机选取15个微卫星（简单序列重复；SSRs）进行测序、特征分析，并基于多位点微卫星分型（MLMT）应用于对来自12个不同国家的87株临床和环境新型隐球菌格鲁比变种分离株进行测序。在这15个SSR位点中，有3个（命名为CNG1、CNG2和CNG3）具有多态性，而其余12个SSR位点未显示出变异。多态性微卫星的特异性PCR引物，即CNG1、CNG2和CNG3，仅能从新型隐球菌菌株（新型隐球菌格鲁比变种、新型隐球菌新型变种和AD杂交种）中扩增出这些位点，而不能从格特隐球菌中扩增出，表明存在种特异性关联。这三个多态性微卫星是菌株基因分型、群体遗传分析、流行病学研究的有用标记，可能有助于新型隐球菌所致隐球菌病的诊断。