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Journal of Clinical Microbiology, May 2001, p. 1731-1737, Vol. 39, No. 5
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.5.1731-1737.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Genetic Polymorphism of Aspergillus fumigatus in Clinical Samples from Patients with Invasive Aspergillosis: Investigation Using Multiple Typing Methods

S. Bertout,1 F. Renaud,2 R. Barton,3 F. Symoens,4 J. Burnod,5 M.-A. Piens,6,8 B. Lebeau,5 M.-A. Viviani,6,8 F. Chapuis,7 J.-M. Bastide,1 R. Grillot,5 M. Mallié,1,* and THE EUROPEAN RESEARCH GROUP ON BIOTYPE AND GENOTYPE OF ASPERGILLUSdagger

Laboratoire d'Immunologie et de Parasitologie, MNERT-EA 2413, Université de Montpellier, 34060 Montpellier Cedex 2,1 Centre d'Étude sur le Polymorphisme des Microorganismes, CEPM UMR CNRS-IRD 9926, IRD, 34032 Montpellier Cedex 1,2 Interactions Cellulaires Parasite-Hôte, MENRT-EA 2940 ER CNRS 2014, Université Joseph Fourier, Grenoble,5 Service de Parasitologie-Mycologie Médicale, Univeresité Claude Bernard, Lyon,6 and Laboratoire d'Information Médicale, Université Lyon, Lyon,7 France; Istituto di Igiene e Medecina Preventiva, Universita degli Studi di Milano, IRCCS Ospedale Maggiore, Milan, Italy8; Division of Microbiology, University of Leeds, Leeds, United Kingdom3; and Scientific Institute of Public Health, Brussels, Belgium4

Received 13 November 2000/Returned for modification 11 January 2001/Accepted 13 February 2001

The genotypes of 52 strains of Aspergillus fumigatus isolated from 12 patients with invasive aspergillosis were investigated using three typing methods (random amplified polymorphic DNA, sequence-specific DNA polymorphism, and microsatellite polymorphism) combined with multilocus enzyme electrophoresis. Isolates were from patients hospitalized in three different geographic areas (Lyon, France; Grenoble, France; and Milan, Italy). In each case, the genetic polymorphism of several colonies (two to five) within the first respiratory clinical sample was studied. For the 52 isolates tested, random amplified polymorphic DNA identified 8 different genotypes, sequence-specific DNA polymorphism identified 9 different types, and microsatellite polymorphism identified 14 types. A combination of these results with multilocus enzyme electrophoresis study identified 25 different types within the sample studied. We identified 3 patients (of the 12 studied) who carried a single genotype; 6 patients were infected by two genotypes, 1 patient had four genotypes, while the last patient had five. A combination of typing methods provided better discrimination than the use of a single method. Typing methods revealed a population structure within each geographical site, suggesting that the epidemiology of A. fumigatus should be considered separately for each of these geographic areas. This study demonstrates the usefulness of combining several typing methods in reaching an understanding of the epidemiology of A. fumigatus and clarifies whether it is sufficient to type one isolate from each specimen to determine the strain involved in invasive aspergillosis.


* Corresponding author. Mailing address: Laboratoire d'Immunologie et de Parasitologie, EA-MENRT 2413, Faculté de Pharmacie 15, Av. Charles Flahault, 34060 Montpellier Cedex 2, France. Phone: 33-4-67-63-52-02. Fax: 33-4-67-41-16-17. E-mail: mmallie{at}iup.pharma.univ-montp1.fr.

dagger European Research Group on Biotype and Genotype of Aspergillus members are as follows: R. Grillot (Project Leader), B. Lebeau, and J. Burnod, Interactions Cellulaires Parasite-Hôte, MENRT-EA 2940 ER CNRS 2014, Université Joseph Fourier, Grenoble, France; N. Nolard, F. Symoens, K. Goens, and S. Heinemann, Scientific Institute of Public Health-Louis Pasteur, Mycology Section, Brussels, Belgium; J. M. Bastide, M. Mallié, S. Bertout, D. Castel, F. Renaud, and T. de Meëus, Laboratoire d'Immunologie et Parasitologie, MNERT-EA 2413, Université de Montpellier, Montpellier, France; M.-A. Piens, E. Dannaoui, M. Perraud, and M.-F. Monier, Service de Parasitologie-Mycologie Médicale, and F. Chapuis, Laboratoire d'Information Médicale, Université Claude Bernard, Lyon, France; M.-A. Viviani, A.-M. Tortorano, M. Cogliati, and A. L. Rigoni, Istituto di Igiene e Medecina Preventiva, Universita degli Studi di Milano, IRCCS Ospedale Maggiore, Milan, Italy; R. Barton, E. G. V. Evans, and H. R. Ashbee, Division of Microbiology, University of Leeds, Leeds, United Kingdom; J. F. Meis, A. Voss, P. E. Verweij, and J. P. Donnelly, Department of Medical Microbiology, University of Nijmegen, Nijmegen, The Netherlands; and P. M. Rath and R. Ansorg, Institut für Medizinische Mikrobiologie, Universität Essen, Essen, Germany.


Journal of Clinical Microbiology, May 2001, p. 1731-1737, Vol. 39, No. 5
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.5.1731-1737.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.



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