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Journal of Clinical Microbiology, September 1998, p. 2413-2418, Vol. 36, No. 9
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Microsatellite Markers for Typing Aspergillus fumigatus Isolates

Emmanuelle Bart-Delabesse,1 Jean-François Humbert,2 Éric Delabesse,3 and Stéphane Bretagne1,*

Laboratoire de Parasitologie-Mycologie, Hôpital Henri Mondor, Créteil,1 Institut National de Recherche Agronomique, Station d'Hydrobiologie Lacustre, Thonon-les-Bains,2 and Laboratoire d'Hématologie, CNRS URA 1461, Hôpital Necker, Paris,3 France

Received 16 March 1998/Returned for modification 22 April 1998/Accepted 1 June 1998

The use of microsatellites as highly polymorphic DNA markers for the typing of isolates of Aspergillus fumigatus was investigated. Four CA repeats were selected by screening an A. fumigatus DNA library with a (CA)10 oligonucleotide. Primers flanking these CA repeats were designed to amplify each locus. One primer of each pair was labeled with a fluorophore, and the PCR products were analyzed with an automatic sequencer and the GeneScan software. For each primer set and for a given isolate, one band was detected and was assigned to an allele because A. fumigatus is haploid. With 50 clinical isolates, 50 environmental isolates, and 2 reference strains we obtained 12, 11, 10, and 23 different alleles for the four CA microsatellites, respectively (discriminatory power, 0.994). The results were identical by whatever DNA extraction technique was used. Interestingly, no clustering between environmental and clinical isolates was observed, suggesting that every isolate is potentially pathogenic. Microsatellite markers appear suitable for use in large epidemiological studies of invasive aspergillosis.

* Corresponding author. Mailing address: Laboratoire de Parasitologie-Mycologie, Hôpital Henri Mondor, 51 avenue du Général DeLattre de Tassigny, 94010, Créteil Cedex, France. Phone: 33 1 49 81 36 41. Fax: 33 1 49 81 36 01. E-mail: bretagne{at}univ-paris12.fr.

Journal of Clinical Microbiology, September 1998, p. 2413-2418, Vol. 36, No. 9
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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