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Journal of Clinical Microbiology, June 1999, p. 1846-1851, Vol. 37, No. 6
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Rapid Identification of Fungi by Using the ITS2 Genetic Region and an Automated Fluorescent Capillary Electrophoresis System

Christine Y. Turenne,1,2,* Steven E. Sanche,3 Daryl J. Hoban,1,4 James A. Karlowsky,1,2,4 and Amin M. Kabani1,2,5

Department of Medical Microbiology, Faculty of Medicine,1 and Faculty of Pharmacy,4 University of Manitoba, and Departments of Clinical Microbiology2 and Medicine,5 Health Sciences Centre, Winnipeg, Manitoba R3A 1R9, Canada, and Division of Infectious Diseases, Royal University Hospital, Saskatoon, Saskatchewan,3 Canada

Received 28 December 1998/Returned for modification 26 January 1999/Accepted 17 March 1999

Invasive fungal disease often plays an important role in the morbidity and mortality of immunocompromised patients. The poor sensitivity of current fungal blood culture and histological practices has led to the development of highly sensitive and specific molecular techniques, such as the PCR. Sequence variability of the internal transcribed spacer 2 (ITS2) region of fungi is potentially useful in rapid and accurate diagnosis of clinical fungal isolates. PCR with fungus-specific primers targeted toward conserved sequences of the 5.8S and 28S ribosomal DNA (rDNA) results in amplification of the species-specific ITS2 regions, which are variable in amplicon length. We have made use of the ABI PRISM 310 genetic analyzer and the ABI PRISM 310 GeneScan analysis software for the determination of variable size differences of the ITS2 region of clinically important fungi, including Candida and non-Candida yeasts, Aspergillus species, and a variety of dermatophytes. No cross-reaction occurred when samples were tested against human and bacterial genomic DNA. We have found that most clinically significant fungal isolates can be differentiated by this method, and it therefore serves to be a promising tool for the rapid (<7 h) diagnosis of fungemia and other invasive fungal infections.

* Corresponding author. Mailing address: Department of Clinical Microbiology, Health Sciences Centre, MS6-820 Sherbrook Street, Winnipeg, Manitoba R3A 1R9, Canada. Phone: (204) 787-4696. Fax: (204) 787-4699. E-mail: cturenne{at}hc-sc.gc.ca.

Journal of Clinical Microbiology, June 1999, p. 1846-1851, Vol. 37, No. 6
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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