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Journal of Clinical Microbiology, February 2000, p. 586-590, Vol. 38, No. 2
Medizinische Klinik, Abteilung
II,1 and
Hygieneinstitut,2 Abteilung Medizinische
Mikrobiologie, Eberhard-Karls-Universität, 72076 Tübingen, Germany
Received 6 August 1999/Returned for modification 14 October
1999/Accepted 10 November 1999
The Light Cycler technique combines rapid in vitro amplification of
DNA in glass capillaries with real-time species determination and
quantification of DNA load. We have established a quantitative PCR
protocol for two clinically important pathogens, Candida
albicans and Aspergillus fumigatus. The sensitivity
of the assay was comparable to those of previously described PCR
protocols (5 CFU/ml). Specific detection of C. albicans and
A. fumigatus could be achieved. The assay showed a high
reproducibility of 96 to 99%. The assay was linear in a range between
101 and 104 Aspergillus conidia. As
capillaries do not have to be reopened for post-PCR analysis, the risk
of carryover contaminations could be minimized. The Light Cycler
allowed quantification of the fungal loads in a limited number of
clinical specimens from patients with hematological malignancies and
histologically proven invasive fungal infections. Five of nine positive
samples had fungal loads between 5 and 10 CFU/ml of blood, two of nine
positive samples had fungal loads between 10 and 100 CFU/ml of blood,
and two of nine samples had fungal loads of more than 100 CFU/ml of
blood. All samples were also found to be PCR positive by
PCR-enzyme-linked immunosorbent assay analysis.
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Quantification of Fungal DNA by Using Fluorescence
Resonance Energy Transfer and the Light Cycler System
*
Corresponding author. Mailing address: Medizinische
Klinik, Abt. II, Labor Prof. Dr. med. H. Einsele, Otfried-Mueller-Str. 10, 72076 Tuebingen, Germany. Phone: 49 7071 2987355. Fax: 49 7071 293179. E-mail: juergen.loeffler{at}med.uni-tuebingen.de.
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