title: Detection and identification of probable endemic fungal pathogen, Cryptococcus gattii, and worldwide pathogen, Cryptococcus neoformans, by real-time PCR.
abstract: A real-time PCR method for detection and identification of Cryptococcus neoformans and Cryptococcus gattii was developed and evaluated using DNA from single-colony or koala nasal smears. Two TaqMan minor groove binder probes that distinguished between these species were designed corresponding to the internal sequences of the CAP59 gene for both species. The real-time PCR assay had 100% specificity, as assessed using 13 reference strains and 300 environmental strains. Twelve smear samples from healthy koalas were analyzed by direct real-time PCR. This method successfully detected C. gattii and C. neoformans in one and three koalas, respectively.
authors.affiliation: Teikyo University Institute of Medical Mycology, Graduate School of Medicine and Faculty of Medicine, Teikyo University, 359 Otsuka, Hachioji, Tokyo 192-0395, Japan.
authors.forename: Kazuo
authors.initials: K
authors.lastname: Satoh
authors.forename: Mari
authors.initials: M
authors.lastname: Maeda
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Umeda
authors.forename: Yoshiharu
authors.initials: Y
authors.lastname: Miyajima
authors.forename: Koichi
authors.initials: K
authors.lastname: Makimura
mesh_terms: D000818:Animals; D056285:Cryptococcus gattii / Q000235:genetics / Q000302:isolation & purification*; D003455:Cryptococcus neoformans / Q000235:genetics / Q000302:isolation & purification*; D004783:Environmental Microbiology; D005656:Fungal Proteins / Q000235:genetics; D009172:Mycology / Q000379:methods*; D009297:Nasal Mucosa / Q000382:microbiology; D015345:Oligonucleotide Probes / Q000235:genetics; D048368:Phascolarctidae / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: C087781:CAP59 protein, Cryptococcus neoformans; D005656:Fungal Proteins; D015345:Oligonucleotide Probes