J Bacteriol Virol.  2013 Jun;43(2):111-119. 10.4167/jbv.2013.43.2.111.

Molecular Typing of Cryptococcus neoformans Isolated from Korean Patients

• Park SH
• Cho SI
• Hwang SM

• KMID: 2168680
• DOI: http://doi.org/10.4167/jbv.2013.43.2.111

Abstract

Cyptococcosis is generally caused by Cryptococcus neoformans, the opportunistic agent which has two species such as C. neoformans and C. gattii. Both C. neoformans and C. gattii species contain a number of genetically diverse subgroups that can be differentiated by various molecular typing methods. We conducted a molecular epidemiological analysis of 30 clinical isolates of the C. neoformans from cryptococcosis patients who had been hospitalized between 2008 and 2010 in medical centers located in Seoul and Busan in Korea. To determine the genetic diversity, 30 strains of C. neoformans were typed using PCR fingerprinting with the microsatellite specific primer of the phage M13 and the restriction fragment length polymorphism (RFLP) of orotidine monophosphosphate pyrophosphorylase (URA5) gene. All isolates were identified as serotype A, mating type MATa and molecular type VNI. The random amplified polymorphic DNA (RAPD) profiles obtained by using two primers revealed a single pattern. Our study shows that 30 strains of clinical C. neoformans are genetically homogeneous, with all of the isolates were molecular type VN1, serotype A, mating type MATa.

Keyword

Cryptococcus neoformans; Molecular type; PCR-fingerprinting; URA5-RFLP; RAPD

MeSH Terms

Bacteriophage M13
Cryptococcosis
Cryptococcus
Cryptococcus neoformans
Dermatoglyphics
DNA
Genetic Variation
Humans
Korea
Microsatellite Repeats
Molecular Typing
Polymerase Chain Reaction
Polymorphism, Restriction Fragment Length
Uridine
DNA
Uridine

Figure

• 

  Figure 1. PCR-fingerprint profiles amplified with the primer M13. Lane #1∼#4, C. neoformans molecular type reference strains VNI, VNII, VNIII, and VNIV; Lane #5∼#16, selected clinical strains (sh98, sh99, sh100, sh107, sh108, sh109, sh110, sh111, sh104, sh105, sh106, and sh118); M, molecular marker (100 bp DNA ladder).

• 

  Figure 2. URA5-RFLP profiles identified after double digestion with the restriction enzymes HhaI and Sau 96I. Lane #1∼#4, C. neoformans molecular type reference strains VNI, VNII, VNIII, and VNIV; Lane #5∼#16, selected clinical strains (sh98, sh99, sh100, sh107, sh108, sh109, sh110, sh111, sh104, sh105, sh106, and sh118); M, molecular marker (100 bp DNA ladder).

• 

  Figure 3. RAPD profiles amplified with the primer OPH-02 (A) and OPH-12 (B). Lane #1, C. neoformans molecular type reference strains VNI; Lane #2∼#16, selected clinical strains (sh98, sh99, sh100, sh107, sh108, sh109, sh110, sh111, sh112, sh114, sh115, sh104, sh105, sh106, and sh118); M, molecular marker (100 bp DNA ladder).

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