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Typing of Scedosporium apiospermum by multilocus enzyme electrophoresis and random amplification of polymorphic DNA

RACHID ZOUHAIR^*,||, ALAIN DEFONTAINE^*, CECILE OLLIVIER, BERNARD CIMON, FRANCOISE SYMOENS, JEAN-NOEL HALLET^*, JEAN DEUNFF and JEAN-PHILIPPE BOUCHARA

*Laboratoire de Biotechnologie, Unité de Biocatalyse, FRE CNRS 2230, Nantes, France, Laboratoire de Parasitologie Pharmaceutique, UMR CNRS 6553, Rennes, France, Laboratoire de Parasitologie-Mycologie, Groupe d'Etude des Interactions Hôte-Parasite, UPRES-EA 3142, Centre Hospitalier Universitaire, Angers, France, Institut Scientifique de Santé Publique-Louis Pasteur, Section Mycologie, Bruxelles, Belgium

Corresponding author: Dr J-P Bouchara (e-mail: Jean-Philippe.Bouchara{at}univ-angers.fr) Present address: Département de Biologie, Faculté des Sciences, Université Moulay Ismail, Meknes, Morocco.

Received 24 Jan. 2001; revised version accepted 19 March 2001.

Abstract

The genetic diversity among epidemiologically unrelated strains of"the human pathogenic fungus Scedosporium apiospermum or its teleomorph, Pseudallescheria boydii, from different areas in Europe, was investigated by multilocus enzyme electrophoresis (MLEE) and random amplification of polymorphic DNA (RAPD). Fourteen enzyme activities were analysed by starch gel electrophoresis, corresponding to 27 polymorphic loci and 43 iso-enzymes. Among the enzymes studied, propionate esterase, carboxyl esterase, superoxide dismutase, carbonate dehydratase and malate dehydrogenase were the most polymorphic, allowing the classification of the strains into 6–11 groups each. Combination of the data obtained for the different enzyme activities studied allowed differentiation of the strains. Similarly, a high polymorphism was also revealed by each of the 20 RAPD primers tested, but no single primer was able to differentiate all the strains. The most efficient primers were GC70, UBC-701 and UBC-703, which revealed 17 distinct genotypes each, and combination of the results obtained with this three-primer set allowed complete discrimination of the strains. The dendrograms obtained from MLEE or RAPD by the unweighted pair-group method using arithmetic average cluster analysis did not reveal any clustering according to the geographic origin of the strains or their pathogenicity.

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