A DNA-based method for the early detection and identification of decay basidiomycetes on wood chips is described. From 26 species of wood-decaying basidiomycetes and 20 species of wood-inhabiting ascomycetes, DNA fragments containing the internal transcribed spacers, 5.8S and partial 28S rDNA were amplified by PCR using the ITS1-F-NL2 rDNA primer pair and analysed through sequencing and restriction digestion. A decay basidiomycete-specific restriction fragment length polymorphism (RFLP) pattern was generated by DraI. Using this PCR-RFLP method the detection of decay fungi is possible 4 days after the inoculation of the wood chips with either a single culture or mixed species.