Journal of Clinical Microbiology · 2007 · 66 citations · 15 references
Life SciencesMicrobiologyMolecular BiologyDiagnostic MicrobiologyMedical MicrobiologyMycologyFungal DiagnosticsFungal GeneticsFungal IdentificationFungal InfectionsFungal PathogenesisPathogen DetectionAntifungal Susceptibility TestingFungal Pathogen DetectionGel ElectrophoresisInternal Transcribed Spacer SequencingMolecular Diagnostics Of Fungal InfectionsPathogen DiagnosisPhenotypic Characterization Of IsolatesPulsed-field Gel Electrophoresis
Data on clinical isolates of Kodamaea (Pichia) ohmeri, an emerging fungal pathogen, are scarce. Over the past 5 years, we identified yeast isolates from nine patients with fungemia as K. ohmeri by using the API 20C system. Here, we reanalyzed these isolates first by sequencing the internal transcribed spacer 2 (ITS2) regions and then by growing the isolates on CHROMagar Candida medium and subjecting them to pulsed-field gel electrophoresis (PFGE). Based on their ITS2 sequences, six of the nine isolates were confirmed as K. ohmeri, while the others were identified as Candida haemulonii (n = 2) and Candida parapsilosis (n = 1). PFGE karyotyping of the K. ohmeri isolates revealed similar major bands, and their colonies showed a characteristic color change from pink to blue when grown on CHROMagar Candida medium for more than 48 h. For K. ohmeri, the ranges of MICs of fluconazole, voriconazole, caspofungin, and micafungin were 2 to 32 mug/ml, 0.03 to 0.5 mug/ml, 0.125 to 0.25 mug/ml, and 0.03 to 0.06 mug/ml, respectively. Restriction endonuclease analysis of genomic NotI-digested DNA (REAG-N) from isolates from different patients produced unique patterns, suggesting that the fungemia had occurred sporadically. This study determined that ITS2 sequence data, PFGE karyotypes, and CHROMagar Candida chromogenic culture medium are reliable diagnostic tools for identifying K. ohmeri while REAG-N is useful for genotyping the clinical isolates of K. ohmeri.
15
PCR protocols — A guide to methods and applications
Trends in Genetics · 1990 · 11.9K citations
Y. C. Chen, J Eisner, Mireille M. Kattar et al. · Journal of Clinical Microbiology · 2000 · 260 citations · Full text
Frank C. Odds, Mary Motyl, Roberto Andrade et al. · Journal of Clinical Microbiology · 2004 · 173 citations · Full text