Różnicowanie izolatów Malassezia pachydermatis metodą RAPD

The aim of the work was analyzing of genomic DNA of Malassezia pachydermatis isolates from clinical cases otitis externa from dogs using RAPD method with arbitrary primers Eric1R, Eric2, BG2 and FM1. Materials and methods. 47 strains of M. pachydermatis isolates from clinical cases otitis externa from dogs were tested. Isolation of genomic DNA was provided according with MasterPureTM Yeast DNA Purification Kit EPICENTRE procedure. The quality of isolated genomic DNA was determined electrophoreticaly. For differentiation the following primers were used: Eric1R, Eric2, BG2 and FM1. Primers Eric 1R and Eric 2 were used together in one reaction or amplificated separately. Obtained products were analyzed electrophoreticaly in 1.5% agarose gel. For determination of phylogenic tree Quantity one VersaDoc (BioRad) and Statgraphics plus 4.1 programs were used. Results. High degree of heterogeneity of DNA among investigated isolates of M. pachydermatis was shown using FM1 primer. Dendrograms were prepared by calculation euclid's distance of different parameters (size and count of RAPD products) by nearest neighbor method. Basing on phylogenic tree four main types (phylogenic groups) of M. pachydermatis isolates were shown. The other five groups non-count was shown also.