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J Med Microbiol 52 (2003), 229-238; DOI: 10.1099/jmm.0.05049-0
© 2003 Society for General Microbiology
ISSN 0022-2615


DIAGNOSTICS, TYPING AND IDENTIFICATION

Detection of seven Candida species using the Light-Cycler system

P. Lewis White, Anjali Shetty and Rosemary A. Barnes

Department of Medical Microbiology and PHLS, University Hospital Wales, Heath Park, Cardiff CF14 4XN, UK

Correspondence P. Lewis White lewis.white{at}phls.wales.nhs.uk


Received 16 August 2002 Accepted 12 November 2002

Due to the limitations of classical methods for the detection of systemic fungal infections and the high mortality rates associated with these infections, it has become essential to develop a quick, sensitive and specific detection assay. By using the Idaho Technologies Light-Cycler system, a qualitative real-time PCR system has been developed for the detection of the leading causes of systemic infection within the genus Candida. The sensitivity of the assay was comparable to previously described PCR methods (1–5 c.f.u. ml-1) and, by the use of a single Candida probe, it was able to detect, but not differentiate between, seven species of Candida (Candida albicans, Candida dubliniensis, Candida glabrata, Candida kefyr, Candida krusei, Candida parapsilosis and Candida tropicalis). Single-round amplification on the Light-Cycler allowed rapid turn-around of clinical samples (within one working day) and it was shown to be more sensitive than classical procedures, exposing 39 possible systemic infections that were not detected by blood culture.


Abbreviations: FRET, fluorescence resonance energy transfer; NASBA, nucleic acid sequence-based amplification; RFLP, restriction fragment length polymorphism.




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