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Evaluation of Pyrosequencing® technology for the identification of clinically relevant non-dematiaceous yeasts and related species

  • C. I. Montero
  • Y. R. Shea
  • P. A. Jones
  • S. M. Harrington
  • N. E. Tooke
  • F. G. Witebsky
  • P. R. Murray
Article

Abstract

Pyrosequencing was used to identify 133 isolates of clinically relevant non-dematiaceous yeasts. These included 97 ATCC strains (42 type strains), seven UAMH strains, and 29 clinical isolates. Isolates belonged to the following genera: Candida (18 species), Trichosporon (10), Cryptococcus (7), Malassezia (3), Rhodotorula (2), Geotrichum (1), Blastoschizomyces (1), and Kodamaea (1). Amplicons of a hyper-variable ITS region were obtained and analyzed using Pyrosequencing technology. The data were evaluated by a BLAST search against the GenBank database and correlated with data obtained by conventional cycle sequencing of the ITS1–5.8S–ITS2 region. Cycle sequencing identified 78.9% of the isolates to the species level. Pyrosequencing technology identified 69.1%. In 90.1% of all of the strains tested, the identification results of both sequencing methods were identical. Most Candida isolates can be identified to the species level by Pyrosequencing. Trichosporon species and some Cryptococcus species cannot be differentiated at the species level. Pyrosequencing can be used for the reliable identification of most commonly isolated non-dematiaceous yeasts, with a reduction of cost per identification compared to conventional sequencing.

Keywords

Clinical Isolate Invasive Fungal Infection ITS2 Region Conventional Sequencing Pyrosequencing Analysis 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

We thank Patricia S. Conville, Department of Laboratory Medicine, Warren Grant Magnuson Clinical Center, NIH, for her technical assistance. This research was supported by the Intramural Research Program of the NIH Clinical Center. The views expressed herein are those of the authors and should not be construed as those of the U.S. Department of Health and Human Services.

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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • C. I. Montero
    • 1
  • Y. R. Shea
    • 1
  • P. A. Jones
    • 1
  • S. M. Harrington
    • 2
  • N. E. Tooke
    • 3
  • F. G. Witebsky
    • 1
  • P. R. Murray
    • 1
  1. 1.Microbiology Service, Department of Laboratory Medicine, Clinical Center, National Institutes of HealthU.S. Department of Health and Human ServicesBethesdaUSA
  2. 2.Department of Clinical MicrobiologyAlbany Medical CenterAlbanyUSA
  3. 3.Biotage ABUppsalaSweden

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