J Guillot1, M Deville, M Berthelemy, F Provost, E Guého. 1. Service de Parasitologie-Mycologie, UMR956 INRA-AFSSA-ENVA, Biologie Moléculaire et Immunologie Parasitaires et Fongiques, Ecole Nationale Vétérinaire d'Alfort, Maisons-Alfort, France. j.guillot@vet-alfort.fr
Abstract
AIMS: The present study describes a system based on PCR and restriction endonuclease analysis (REA) to distinguish the seven currently recognized Malassezia species. METHODS AND RESULTS: Fifty-five representative yeast isolates were examined. A single primer pair was designed to amplify the large subunit ribosomal RNA (LSU rRNA) gene of the seven Malassezia species, and identification was achieved by digestion of the PCR products with three restriction endonucleases: BanI, HaeII and MspI. A specific restriction endonuclease analysis pattern was determined for each species investigated. Moreover, PCR-REA allowed the detection and characterization of mixtures of several Malassezia species. CONCLUSION: PCR-REA of only the LSU rRNA gene is a reliable and rapid method to distinguish all Malassezia species. SIGNIFICANCE AND IMPACT OF THE STUDY: PCR-REA represents a considerable saving in time over currently available identification procedures. This method should be evaluated on clinical material directly.
AIMS: The present study describes a system based on PCR and restriction endonuclease analysis (REA) to distinguish the seven currently recognized Malassezia species. METHODS AND RESULTS: Fifty-five representative yeast isolates were examined. A single primer pair was designed to amplify the large subunit ribosomal RNA (LSU rRNA) gene of the seven Malassezia species, and identification was achieved by digestion of the PCR products with three restriction endonucleases: BanI, HaeII and MspI. A specific restriction endonuclease analysis pattern was determined for each species investigated. Moreover, PCR-REA allowed the detection and characterization of mixtures of several Malassezia species. CONCLUSION: PCR-REA of only the LSU rRNA gene is a reliable and rapid method to distinguish all Malassezia species. SIGNIFICANCE AND IMPACT OF THE STUDY: PCR-REA represents a considerable saving in time over currently available identification procedures. This method should be evaluated on clinical material directly.
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