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Journal of Clinical Microbiology, August 2000, p. 2829-2836, Vol. 38, No. 8
Public Health Research
Institute,1 and Memorial Sloan-Kettering
Cancer Center,2 New York, and
Wadsworth Center, New York State Health Department,
Albany,3 New York
Received 6 March 2000/Returned for modification 1 May 2000/Accepted 14 May 2000
Candida dubliniensis is an opportunistic fungal
pathogen that has been linked to oral candidiasis in AIDS patients,
although it has recently been isolated from other body sites. DNA
sequence analysis of the internal transcribed spacer 2 (ITS2) region of rRNA genes from reference Candida strains was used to
develop molecular beacon probes for rapid, high-fidelity identification of C. dubliniensis as well as C. albicans.
Molecular beacons are small nucleic acid hairpin probes that brightly
fluoresce when they are bound to their targets and have a significant
advantage over conventional nucleic acid probes because they exhibit a
higher degree of specificity with better signal-to-noise ratios. When applied to an unknown collection of 23 strains that largely contained C. albicans and a smaller amount of C. dubliniensis, the species-specific probes were 100% accurate in
identifying both species following PCR amplification of the ITS2
region. The results obtained with the molecular beacons were
independently verified by random amplified polymorphic DNA
analysis-based genotyping and by restriction enzyme analysis with
enzymes BsmAI and NspBII, which cleave
recognition sequences within the ITS2 regions of C. dubliniensis and C. albicans, respectively. Molecular
beacons are promising new probes for the rapid detection of
Candida species.
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Rapid Identification of Candida
dubliniensis Using a Species-Specific Molecular
Beacon
*
Corresponding author. Mailing address: Public Health
Research Institute, 455 First Ave., New York, NY 10016. Phone: (212) 578-0820. Fax: (212) 578-0804. E-mail:
perlin{at}phri.nyu.edu.
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