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Related Experiment Videos

Polymorphic internal transcribed spacer region 1 DNA sequences identify medically important yeasts.

Y C Chen1, J D Eisner, M M Kattar

  • 1Department of Laboratory Medicine, University of Washington, Seattle, Washington 98195, USA.

Journal of Clinical Microbiology
|October 30, 2001
PubMed
Summary
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Analyzing internal transcribed spacer 1 (ITS1) and internal transcribed spacer 2 (ITS2) regions accurately identifies clinically significant yeasts. Combining ITS1 and ITS2 polymorphisms reliably identifies 40 yeast species, aiding in diagnosing unusual clinical isolates.

Area of Science:

  • Medical Mycology
  • Molecular Diagnostics
  • Yeast Identification

Background:

  • Accurate identification of clinically significant yeasts is crucial for effective treatment.
  • Noncoding internal transcribed spacer 2 (ITS2) region polymorphisms are established markers for yeast identification.
  • The diagnostic utility of the internal transcribed spacer 1 (ITS1) region remains less explored.

Purpose of the Study:

  • To investigate the diagnostic value of internal transcribed spacer 1 (ITS1) noncoding regions for yeast identification.
  • To compare the discriminatory power of ITS1 and ITS2 regions, individually and in combination.
  • To assess the potential of ITS polymorphisms for identifying novel or unusual yeast pathogens.

Main Methods:

  • Amplification of ITS1 and ITS2 regions using polymerase chain reaction (PCR).

Related Experiment Videos

  • Automated capillary electrophoresis for rapid determination of PCR product length with single-base precision.
  • Restriction enzyme analysis for distinguishing yeasts with similar PCR product sizes.
  • DNA sequencing of ITS1 regions for detailed allelic analysis and phylogenetic comparisons.

    • Main Results:

    • ITS1 length polymorphisms distinguished 19 species, while ITS2 distinguished 16 species.
    • Combining ITS1 and ITS2 analyses identified 30 species (98% of clinical isolates).
    • Restriction enzyme analysis and ITS1 sequencing further resolved remaining species, enabling identification of all 40 target species.
    • Phylogenetic analysis using ITS sequences showed congruence with 26S rRNA gene trees, with ITS providing higher resolution for closely related species.

    Conclusions:

    • Analysis of ITS polymorphisms, particularly when combining ITS1 and ITS2 regions, provides a reliable method for identifying 40 clinically significant yeast species.
    • ITS1 sequencing offers species-specific alleles valuable for identifying unusual clinical isolates.
    • The established database of ITS sequences holds promise for the identification of potentially new pathogenic yeast species.