ArticleJournal of clinical microbiology2000
Identification of medically important yeasts using PCR-based detection of DNA sequence polymorphisms in the internal transcribed spacer 2 region of the rRNA genes.
Article in Journal of clinical microbiology, 2000. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 82 papers.
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Who cites it
82 citing papers in PubMed.
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- Epidemiology and Prevalence of Oral Candidiasis in HIV Patients From Chad in the Post-HAART Era.Frontiers in microbiology · 2022Article
- Host Age and Denture Wearing Jointly Contribute to Oral Colonization with Intrinsically Azole-Resistant Yeasts in the Elderly.Microorganisms · 2021Article
- Chronic incurable mastitis in sheep: prevalence, identification of predisposing factors, and genotyping of fungal causative species using PCR-RFLP.Tropical animal health and production · 2021Article
- The Role of Fatty Acid Metabolites in Vaginal Health and Disease: Application to Candidiasis.Frontiers in microbiology · 2021Review
- Oral Colonization by Candida Species in Patients with Multiple Sclerosis.Mycopathologia · 2020Article
- In vitro antifungal resistance profile of Candida strains isolated from Saudi women suffering from vulvovaginitis.European journal of medical research · 2020Article
- Genomic and transcriptomic analysis ofBiotechnology for biofuels · 2020Article
- Hyper-production of taxol fromBiotechnology reports (Amsterdam, Netherlands) · 2019Article
- Single-tube, dual channel pentaplexing for the identification of Candida strains associated with human infection.Scientific reports · 2019Article
- Advances in Chemical and Biological Methods to Identify Microorganisms-From Past to Present.Microorganisms · 2019Review
22 more citing papers are in PubMed but not listed here.
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Authors and funding
8 authors.
Funding
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Abstract
Identification of medically relevant yeasts can be time-consuming and inaccurate with current methods. We evaluated PCR-based detection of sequence polymorphisms in the internal transcribed spacer 2 (ITS2) region of the rRNA genes as a means of fungal identification. Clinical isolates (401), reference strains (6), and type strains (27), representing 34 species of yeasts were examined. The length of PCR-amplified ITS2 region DNA was determined with single-base precision in less than 30 min by using automated capillary electrophoresis. Unique, species-specific PCR products ranging from 237 to 429 bp were obtained from 92% of the clinical isolates. The remaining 8%, divided into groups with ITS2 regions which differed by </=2 bp in mean length, all contained species-specific DNA sequences easily distinguishable by restriction enzyme analysis. These data, and the specificity of length polymorphisms for identifying yeasts, were confirmed by DNA sequence analysis of the ITS2 region from 93 isolates. Phenotypic and ITS2-based identification was concordant for 427 of 434 yeast isolates examined using sequence identity of >/=99%. Seven clinical isolates contained ITS2 sequences that did not agree with their phenotypic identification, and ITS2-based phylogenetic analyses indicate the possibility of new or clinically unusual species in the Rhodotorula and Candida genera. This work establishes an initial database, validated with over 400 clinical isolates, of ITS2 length and sequence polymorphisms for 34 species of yeasts. We conclude that size and restriction analysis of PCR-amplified ITS2 region DNA is a rapid and reliable method to identify clinically significant yeasts, including potentially new or emerging pathogenic species.
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