Journal List > J Bacteriol Virol > v.40(2) > 1033986

Hwang: Molecular Characterization of Clinical and Environmental Strains of Cryptococcus neoformans Isolated from Busan, Korea

Abstract

Cryptocococcus neoformans is an encapsulated yeast that can cause life-threatening infections in immunocompromised patients. In this study, the genetic variability and epidemiological relationships of clinical and environmental isolates of C. neoformans from Busan, Korea, 2000~2005 were investigated. A total of 12 strains of C. neoformans, 7 clinical and 5 environmental isolates were analyzed by random amplified polymorphic DNA (RAPD) using three different primers and PCR-fingerprinting with a minisatellite-specific core sequence of phage M13. All strains belonged to C. neoformans serotype A and mating type MATa. Two different RAPD profiles (I and II) and a single pattern by M13 PCR-fingerprinting were identified. The major RAPD profile was pattern I (8 of 12 strains) and pattern II was identified from 2 clinical and 2 environmental strains, which clearly distinguished among isolates. Clinical strains with pattern II were isolated from the patients with HIV positive. Taken together, molecular patterns provide a good characterization of strains of C. neoformans as a heterogeneous group and epidemiological relationships in clinical and environmental strains.

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Figure 1.
PCR products of C. neoformans isolates for determination of mating type α and a. Lane 1~5: clinical and environmental isolates, 6: negative control, M: molecular marker
jbv-40-91f1.tif
Figure 2.
RAPD profiles of C. neoformans isolates amplified with the primers. (A): Primer OPH-02, (B): Primer OPH-12, (C): Primer OPH-20. M: molecular marker.
jbv-40-91f2.tif
Figure 3.
PCR fingerprint profiles amplified with the M13 primer. M: molecular marker.
jbv-40-91f3.tif
Table 1.
Sources, serotypes, and mating types of C. neoformans isolates in Busan, Korea
No. Strain Type of isolates Source Year of isolation Serotype Mating type
1 C21 aC CSF 2000 A a
2 C22 C CSF 2001 A a
3 C28 C Blood 2002 A a
4 C36 C Blood 2003 A a
5 C40 C cCSF 2003 A a
6 C41 C Blood 2005 A a
7 C58 C CSF 2004 A a
8 C23 bE Pigeon excreta 2002 A a
9 C24 E Pigeon excreta 2002 A a
10 C25 E Pigeon excreta 2002 A a
11 C26 E Pigeon excreta 2002 A a
12 C27 E Pigeon excreta 2002 A a

a C; clincal source,

b E; environmental source,

c CSF; cerebrospinal fluid

Table 2.
RAPD patterns of C. neoformans isolates from Busan, Korea
Pattern RAPD profile Strain No.
Primer OPH-02 Primer OPH-12 Primer OPH-20
I 1 a A C21 C22 C28 C36 C40 C23 C26 C27
II 2 b B C41 C58 C24 C25
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