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J Med Microbiol 55 (2006), 1211-1216; DOI: 10.1099/jmm.0.46723-0
© 2006 Society for General Microbiology
ISSN 1473-5644

Fast and reliable PCR/sequencing/RFLP assay for identification of fungi in onychomycoses

Michel Monod, Olympia Bontems, Christophe Zaugg, Barbara Léchenne, Marina Fratti and Renato Panizzon

Department of Dermatology, Centre Hospitalier Universitaire Vaudois, CH-1011 Lausanne, Switzerland

Correspondence
Michel Monod
Michel.Monod{at}chuv.ch

Received 15 May 2006
Accepted 30 May 2006

Fusarium spp. and other non-dermatophyte fungi are repeatedly isolated from abnormal nails. To investigate whether these fungi are the aetiological agents of infection or simply transient contaminants, a PCR/sequencing/RFLP assay was developed for direct and routine identification of the infecting fungi in onychomycosis. Fungal DNA was readily extracted using a commercial kit after dissolving nail fragments in a Na2S solution. Amplification of part of the 28S rDNA by PCR was performed with universal primers and the fungal species were identified by sequencing. The PCR/sequencing results were comparable with microbiological identification from the same nail sample. In addition to dermatophytes, Fusarium spp. and other less frequently isolated non-dermatophyte fungi were identified as single fungal agents in onychomycosis. Moreover, mixed infections were clearly demonstrated in 10 % of cases by RFLP analysis of PCR products. Identification of infectious agents could be obtained in 2 days, whilst results from fungal cultures take 1–3 weeks. Rapid and reliable molecular identification of the infectious fungus expedites the choice of appropriate antifungal therapy, thereby improving the cure rate of onychomycosis.

Abbreviations: LSU, large subunit; NDF, non-dermatophyte filamentous fungi.






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