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Journal of Clinical Microbiology, February 1998, p. 395-401, Vol. 36, No. 2
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

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

M. Flahaut,1,2 D. Sanglard,1 M. Monod,3 J. Bille,1 and M. Rossier2,*

Institut de Microbiologie1 and Laboratoire de Mycologie,3 Centre Hospitalier Universitaire Vaudois, Lausanne, and Laboratoires, Hôpital de Zone de Morges, Morges,2 Switzerland

Received 5 June 1997/Returned for modification 1 October 1997/Accepted 4 November 1997

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 SAP genes 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 an SAP-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.

* Corresponding author. Mailing address: Laboratoires, Hôpital de Zone de Morges, Chemin du Cret 2, CH 1110 Morges, Switzerland. Phone: 41 21 804 20 18. Fax: 41 21 804 20 12. E-mail: Marjorie.Flahaut{at}chuv.hospvd.ch.

Journal of Clinical Microbiology, February 1998, p. 395-401, Vol. 36, No. 2
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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