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Journal of Clinical Microbiology, September 2001, p. 3402-3408, Vol. 39, No. 9
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.9.3402-3408.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Colorimetric Assay for Antifungal Susceptibility Testing of Aspergillus Species

Joseph Meletiadis,1 Johan W. Mouton,2 Jacques F. G. M. Meis,3 Bianca A. Bouman,1 J. Peter Donnelly,3 Paul E. Verweij,1,* and EUROFUNG Networkdagger

Departments of Medical Microbiology1 and Hematology,3 University Medical Center Nijmegen, and Department of Medical Microbiology and Regional Public Health Laboratory, Canisius Wilhelmina Hospital,2 Nijmegen, The Netherlands

Received 20 February 2001/Returned for modification 11 April 2001/Accepted 27 June 2001

A colorimetric assay for antifungal susceptibility testing of Aspergillus species (Aspergillus fumigatus, Aspergillus flavus, Aspergillus terreus, Aspergillus nidulans, and Aspergillus ustus) is described based on the reduction of the tetrazolium salt 2,3-bis(2-methoxy-4-nitro-5-[(sulphenylamino)carbonyl]-2H-tetrazolium-hydroxide (XTT) in the presence of menadione as an electron-coupling agent. The combination of 200 µg of XTT/ml with 25 µM menadione resulted in a high production of formazan within 2 h of exposure, allowing the detection of hyphae formed by low inocula of 102 CFU/ml after 24 h of incubation. Under these settings, the formazan production correlated linearly with the fungal biomass and less-variable concentration effect curves for amphotericin B and itraconazole were obtained.

* Corresponding author. Mailing address: Department of Medical Microbiology, University Medical Center Nijmegen, P.O. Box 9101, 6500 HB Nijmegen, The Netherlands. Phone: 31-24-3613514. Fax: 31-24-3540216. E-mail: p.verweij{at}mmb.azn.nl.

dagger EC-TMR-EUROFUNG network (ERBFMXR-CT970145). The Eurofung Network consists of the following participants: Emmanuel Roilides and Nicos Maglaveras, Aristotle University, Thessaloniki, Greece; Tore Abrahamsen and Peter Gaustad, Rikshospitalet National Hospital, Oslo, Norway; David W. Denning, University of Manchester, Manchester, United Kingdom; Paul E. Verweij and Jacques F. G. M. Mcis, University of Nijmegen, Nijmegen, The Netherlands; Juan L. Rodriguez-Tudela, Instituto de Salud Carlos III, Madrid, Spain; and George Petrikkos, Athens University, Athens, Greece.

Journal of Clinical Microbiology, September 2001, p. 3402-3408, Vol. 39, No. 9
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.9.3402-3408.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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