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J. Clin. Microbiol. doi:10.1128/JCM.00514-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Efficient identification of clinically relevant yeasts combining a panfungal loop-mediated isothermal DNA amplification (LAMP) with hybridization to species-specific oligonucleotide probes

Centro de Recursos Microbiológicos (CREM), Department of Life Sciences, Faculty of Sciences and Technology, New University of Lisbon, 2829-516 Caparica, Portugal; STAB Vida Lda., Apartado 89, 2781-601 Oeiras, Portugal

^* To whom correspondence should be addressed. Email: ism{at}fct.unl.pt.

	Abstract

The occurrence of invasive mycoses has progressively increased in recent years. Yeasts of the genus Candida remain the leading etiologic agents of those infections. Early identification of opportunistic yeasts may contribute significantly to improved disease management and the selection of appropriate antifungal therapy. We developed a rapid and reliable molecular identification system for clinically relevant yeasts which makes use of non-specific primers to amplify a region of the 26S rRNA gene, followed by reverse hybridization of the digoxigenin-labelled products to a panel of species-specific oligonucleotide probes arranged on a nylon membrane macroarray format. DNA amplification was achieved by the recently developed Loop-Mediated Isothermal DNA Amplification (LAMP) technology, a promising option for the development of improved laboratory diagnostic kits. The newly developed method was successful in discriminating among the major clinically relevant yeasts associated with bloodstream infections, using simple, rapid and cost-effective procedures and equipment.