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04 May 2007

Significant discrepancy between real-time PCR identification and hospital identification of C. albicans from Lebanese patients

Suha Yazbek, Ghyda Barada, Raida Basma, Jad Mahfouz, Roy A. Khalaf

Med Sci Monit 2007; 13(5): MT7-12 :: ID: 484614

Abstract

Background: The fungus C. albicans is among the leading agents causing death in immunocompromised patients. Most hospitals rely on conventional morphological techniques such as the germ-tube assay
and the API system for correct identifi cation. This technique is subjective and hence error prone. Recently, more and more molecular techniques for correct identifi cation have been developed. The latest is the LightCycler real-time PCR technique coupled with melting curve analysis.
Material/Methods: One hundred hospital isolates presumed to be C. albicans from four major Lebanese hospitals were tested using the real-time PCR technique. The results were compared with a germ-tube test.
Furthermore, all real-time PCR-positive samples were replica plated on growth media at 30°C and 45°C to differentiate between C. albicans and C. dubliniensis. Finally, all PCR-negative samples were identifi ed using the API 20C AUX yeast identifi cation system.
Results: Twenty-four hospital isolates were non-albicans by PCR (ppC. albicans-negative isolates were misidentifi ed by the API 20C AUX. None of the C. albicans real-time PCR-positive samples failed to grow at 45°C, the
dubliniensis non-permissive temperature.
Conclusions: Considering the impact of false identifi cation on the general public health through the use of wrong antifungal drug treatment and the emergence of novel drug-resistant strains, hospitals should update their classifi cation methods using molecular techniques.

Keywords: Candidiasis - drug therapy, Candida albicans - genetics, Antifungal Agents - therapeutic use, Drug Resistance, Fungal, Hospitals, Lebanon, Mycological Typing Techniques, Polymerase Chain Reaction - methods

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