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Journal of Clinical Microbiology, November 2002, p. 4308-4312, Vol. 40, No. 11
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.11.4308-4312.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Rapid Identification and Differentiation of Candida albicans and Candida dubliniensis by Capillary-Based Amplification and Fluorescent Probe Hybridization

Departments of Laboratory Medicine,¹ Medicine,² Microbiology, University of Washington Medical Center, Seattle, Washington 98195-7110³

Received 2 May 2002/ Returned for modification 2 July 2002/ Accepted 4 August 2002

We developed a rapid genotypic assay to differentiate the germ tube-positive yeasts Candida albicans and Candida dubliniensis. Fluorescently labeled nucleic acid probe binding and subsequent denaturation from the target site in the PCR amplicons produced characteristic peak melting temperatures (T_m) that identified each species. Peak T_ms of C. albicans (n = 69) and C. dubliniensis (n = 28) isolates produced in the presence of their respective probes were 61.04 ± 0.64°C and 60.52 ± 1.01°C (averages ± standard deviations). No signal was generated when the C. albicans or C. dubliniensis probes were tested against DNA from their counterparts. Both probes reacted with Candida tropicalis DNA, but the T_m was 51.85 ± 0.05°C with the C. albicans probe and 51.92 ± 0.10°C with the C. dubliniensis probe, differentiating C. tropicalis DNA from C. albicans and C. dubliniensis. A novel hybrid probe was designed to identify both species in a single reaction based on a 4°C difference in peak T_ms. Our assay is rapid (2 h) and allows reliable detection and differentiation of the two germ tube-positive Candida spp.

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