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Journal of Clinical Microbiology
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Mycology

Rapid Detection of Candida albicans in Clinical Samples by DNA Amplification of Common Regions from C. albicans-Secreted Aspartic Proteinase Genes

M. Flahaut, D. Sanglard, M. Monod, J. Bille, M. Rossier
M. Flahaut
Institut de Microbiologie and
Laboratoires, Hôpital de Zone de Morges, Morges, Switzerland
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D. Sanglard
Institut de Microbiologie and
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M. Monod
Laboratoire de Mycologie, Centre Hospitalier Universitaire Vaudois, Lausanne, and
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J. Bille
Institut de Microbiologie and
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M. Rossier
Laboratoires, Hôpital de Zone de Morges, Morges, Switzerland
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DOI: 10.1128/JCM.36.2.395-401.1998
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ABSTRACT

Laboratory diagnosis based on genomic amplification methods such as PCR may provide an alternative and more sensitive method than conventional culture for the early detection of deep-seated candidiasis, an increasing cause of morbidity and mortality among immunocompromised patients. A novel method of DNA extraction from clinical samples based on treatment with proteinase K and isolation of DNA on a silica membrane was developed. The targets used for DNA amplification were the Candida albicans-secreted aspartic proteinase (SAP) genes, a multiple-gene family of at least seven members in C. albicans. A single pair of primers was designed in order to detect six of these SAPgenes and, subsequently, to increase the sensitivity of the test. Detection of the PCR product by enzyme-linked immunosorbent assay was found to be as sensitive as Southern blotting with anSAP-labeled probe. The sensitivity of the assay was 1 cell/ml from serially diluted Candida cultures and 1 to 4 cells/ml from seeded blood specimens. The sensitivity and specificity of the present assay were tested in a retrospective study performed blindly with 156 clinical samples and were 100 and 98%, respectively, compared with the results of culture. For the subset of blood culture samples (n = 124), the sensitivity and the specificity were 100%. The two false-positive PCR samples came from patients treated with azole antifungal agents, indicating that PCR was probably able to detect damaged organisms that could not be recovered by culture.

  • Copyright © 1998 American Society for Microbiology
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Rapid Detection of Candida albicans in Clinical Samples by DNA Amplification of Common Regions from C. albicans-Secreted Aspartic Proteinase Genes
M. Flahaut, D. Sanglard, M. Monod, J. Bille, M. Rossier
Journal of Clinical Microbiology Feb 1998, 36 (2) 395-401; DOI: 10.1128/JCM.36.2.395-401.1998

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Rapid Detection of Candida albicans in Clinical Samples by DNA Amplification of Common Regions from C. albicans-Secreted Aspartic Proteinase Genes
M. Flahaut, D. Sanglard, M. Monod, J. Bille, M. Rossier
Journal of Clinical Microbiology Feb 1998, 36 (2) 395-401; DOI: 10.1128/JCM.36.2.395-401.1998
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KEYWORDS

Aspartic Acid Endopeptidases
Candida albicans
candidiasis
DNA, Fungal
polymerase chain reaction

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