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Molecular identification of fungi of clinical relevance

AuthorsUnda, F.; Martínez-Martínez, Luis
Issue DateApr-2010
CitationECCMID (2010)
Abstract[Objectives]}: Culture and conventional identification remain the usual basis for diagnosis of fungal infections, but they have a long response time and a low sensitivity rate. The objectives of this study were to perform molecular identification of clinically relevant fungi not identified at the species level with a conventional approach (culture, API ID32C and morphologic criteria) and to compare the results of molecular detection of fungi in invasive samples (other than blood) in those of culture. [Methods]: In a preliminary phase, 10 strains of international collections were tested. After they were all correctly recognized, two prospective studies were done in parallel during one year (Sept. 2008 to Aug. 2009): (i) 36 cultures of difficult-to-identify fungi (12 yeasts and 24 moulds) obtained from different clinical samples were identified by molecular methods, and (ii) the presence and identification of fungi in 39 invasive samples other than blood (10 bronchoalveolar lavages, 5 bronchoalveolar brushes, 7 heart valves and 17 biopsies (joint: 8;lung: 7;brain: 2) were determined using a conventional approach and compared with the results of a molecular method based on sequencing the Internal Transcriber Spacer (ITS) regions 1 and 2, complemented with sequencing the beta-tubulin and the elongation factor genes, and the intergenic spacer (IGS) region. [Results]: All 36 organisms of objective (i) were identified by the molecular method as concrete species of genera Aspergillus (9), Candida (7), Trichosporon (5), Scedosporium (3), Alternaria (3), fusarium (3), Microsporum (2), Penicilium (2), Sporotrichum (1) or Acremonium (1). 35 out the 39 samples of objective (ii) were negative by both culture and molecular methods. Moulds were identified by the molecular approach in two cases in which the same organism grew in culture (a joint biopsy and a heart valve from the same patient yielding Scedosporium apiospermum). Additionally, the molecular approach identified an Aspergillus sidowii in a lung biopsy and an A. fumigatus in a bronchoalveolar brush in two culture-negative cases. The molecular method allowed identification of the organism (from either culture or clinical samples) in 48-72 hours. [Conclusions]: Molecular methods reduces the response time for identification of clinically relevant fungi (include those for which conventional identification is difficult). This approach should be considered for the diagnosis of fungal infections in the clinical laboratory.
DescriptionComunicación presentada al "20th European Congress of Clinical Microbiology and Infectious Diseases" celebrado en Viena (Austria) del 10 al 13 de abril de 2010.-- et al.
Publisher version (URL)http://www.congrex-switzerland.com/eccmid2010/
Appears in Collections:(IBBTEC) Comunicaciones congresos
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