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Medical Mycology Volume 46, 2008 - Issue 7
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Original Article

Multilocus microsatellite typing for Cryptococcus neoformans var. grubii

Ahmed Hanafy Medical Mycology Research Center (former Research Center for Pathogenic Fungi and Microbial Toxicoses), Chiba University, Chiba, Japan
,
Sirada Kaocharoen Molecular Mycology Research Laboratory, Westmead Millennium Institute, Department of Medicine, University of Sydney Western Clinical School, ICPMR, CIDM, Westmead Hospital, Westmead, New South Wales, Australia; Mycology Unit, Department of Microbiology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand
,
Alejandro Jover-Botella Molecular Mycology Research Laboratory, Westmead Millennium Institute, Department of Medicine, University of Sydney Western Clinical School, ICPMR, CIDM, Westmead Hospital, Westmead, New South Wales, Australia
,
Masakazu Katsu Medical Mycology Research Center (former Research Center for Pathogenic Fungi and Microbial Toxicoses), Chiba University, Chiba, Japan
,
Soji Iida Medical Mycology Research Center (former Research Center for Pathogenic Fungi and Microbial Toxicoses), Chiba University, Chiba, Japan
,
Takahisa Kogure Medical Mycology Research Center (former Research Center for Pathogenic Fungi and Microbial Toxicoses), Chiba University, Chiba, Japan
,
Tohru Gonoi Medical Mycology Research Center (former Research Center for Pathogenic Fungi and Microbial Toxicoses), Chiba University, Chiba, Japan
,
Prof Yuzuru Mikami Medical Mycology Research Center (former Research Center for Pathogenic Fungi and Microbial Toxicoses), Chiba University, Chiba, JapanCorrespondence[email protected]
&
Wieland Meyer Molecular Mycology Research Laboratory, Westmead Millennium Institute, Department of Medicine, University of Sydney Western Clinical School, ICPMR, CIDM, Westmead Hospital, Westmead, New South Wales, Australia
 show all
Pages 685-696 | Received 04 Apr 2007, Published online: 09 Jul 2009
 

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.

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