APPLICABILITY OF RANDOM PRIMER R143 FOR DETERMINATION OF ASPERGILLUS-FUMIGATUS DNA

The specificity of random primer R143 for Aspergillus fumigatus DNA wa s determined in order to test its usefulness in establishing the prese nce of A. fumigatus DNA in fungal cultures. When PCR reaction products of these cultures were compared with those of 21 other bacterial and fungal DNA samples, R143 proved to produce a 1346 bp band with only A. fumigatus. This band has been sequenced completely and the EcoRI rest riction site was used for subsequent confirmation of PCR products. The specificity for A. fumigatus DNA was also confirmed by Southern blott ing. Comparison of morphological typing of Aspergillus species in cult ures with PCR using R143 on DNA isolated from these cultures showed co ncordance in 22 of 24 cases. In two cases there was discordance: both times PCR results showed correctly the presence of A. fumigatus, initi ally not detected by culture. R143 is an A. fumigatus specific random primer, with potential for use in detection of A. fumigatus DNA in cli nical specimens.