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Journal of Clinical Microbiology, April 2007, p. 1152-1158, Vol. 45, No. 4
0095-1137/07/$08.00+0     doi:10.1128/JCM.02061-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Molecular Identification of Unusual Pathogenic Yeast Isolates by Large Ribosomal Subunit Gene Sequencing: 2 Years of Experience at the United Kingdom Mycology Reference Laboratory{triangledown}

Christopher J. Linton,1* Andrew M. Borman,1 Grace Cheung,2 Ann D. Holmes,1 Adrien Szekely,1 Michael D. Palmer,1 Paul D. Bridge,3 Colin K. Campbell,1 and Elizabeth M. Johnson1

Mycology Reference Laboratory, Health Protection Agency, Bristol,1 Department of Pathology and Microbiology, University of Bristol, Bristol,2 British Antarctic Survey, Cambridge, United Kingdom3

Received 6 October 2006/ Returned for modification 7 November 2006/ Accepted 13 January 2007

Rapid identification of yeast isolates from clinical samples is particularly important given their innately variable antifungal susceptibility profiles. We present here an analysis of the utility of PCR amplification and sequence analysis of the hypervariable D1/D2 region of the 26S rRNA gene for the identification of yeast species submitted to the United Kingdom Mycology Reference Laboratory over a 2-year period. A total of 3,033 clinical isolates were received from 2004 to 2006 encompassing 50 different yeast species. While more than 90% of the isolates, corresponding to the most common Candida species, could be identified by using the AUXACOLOR2 yeast identification kit, 153 isolates (5%), comprised of 47 species, could not be identified by using this system and were subjected to molecular identification via 26S rRNA gene sequencing. These isolates included some common species that exhibited atypical biochemical and phenotypic profiles and also many rarer yeast species that are infrequently encountered in the clinical setting. All 47 species requiring molecular identification were unambiguously identified on the basis of D1/D2 sequences, and the molecular identities correlated well with the observed biochemical profiles of the various organisms. Together, our data underscore the utility of molecular techniques as a reference adjunct to conventional methods of yeast identification. Further, we show that PCR amplification and sequencing of the D1/D2 region reliably identifies more than 45 species of clinically significant yeasts and can also potentially identify new pathogenic yeast species.

* Corresponding author. Mailing address: Mycology Reference Laboratory, Health Protection Agency South-West Regional Laboratory, Myrtle Road, Bristol BS2 8EL, United Kingdom. Phone: 117 926 8683. Fax: 117 922 6611. E-mail: Chris.Linton{at}bris.ac.uk

{triangledown} Published ahead of print on 24 January 2007.

Journal of Clinical Microbiology, April 2007, p. 1152-1158, Vol. 45, No. 4
0095-1137/07/$08.00+0     doi:10.1128/JCM.02061-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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