Specific detection of Amanita phalloides mycelium and spores by PCR amplification of the GPD (glyceraldehyde-3-phosphate dehydrogenase) gene fragment

Oligonucleotide primers designed to flank a 635 bp fragment of the gene enc oding glyceraldehyde-3-phosphate dehydrogenase (gpd) from Amanita muscaria were used to amplify the corresponding gpd fragment from Amanita phalloides . The A. phalloides PCR product was cloned, sequenced and found to be 70 - 77% similar to the known basidiomycetes gpd genes within the exon part and 25 - 52% within the intron part. Based on these data, species-specific ampl ification was achieved using a pair of oligonucleotide primers complementar y to the A. phalloides gpd intron sequences. These primers allowed the ampl ification of the corresponding gpd fragment from the A. phalloides but not from various other basidiomycetes, ascomycetes and human matrices. PCR ampl ification of the A. phalloides DNA gave the predicted PCR product of 284 bp . The created PCR system is an efficient tool for the specific, rapid and s ensitive detection of A. phalloides mycelium and spores.