Conventional techniques for identification and typing of filamentous fungi are time consuming and laborious. As infections with especially A.fumigatus are seen with increasing frequency, faster and easier recognition methods of this fungus are needed. In order to achieve this goal we evaluated RAPD PCR technique for identification of A.fumigatus. For that purpose we have developed a 20mer primer (R5), which sequence was based on a product raised by 10-mer RAPD R143 primer and A.fumigatus DNA. According to the report from Aufavre et al. PCR with R143 primer gives a single product with A.fumigatus and none with 5 other Aspergilli. PCR with our 20-mer R5 primer and DNA from eight different aspergilli species (A.flavus, A.fumigatus, A.nidulans, A.niger, A.amstelodami, A.terreus, A.Ustus, A.versicolor, seven other pathogenic fungi (Pseudoallecheria boydii, Fusarium spec., Paecilomyces variotii, P.chrysogenum, p.marneffei, C.neoformans, Rhizopus spec.), five yeasts (C.albicans, C.krusei, C.tropicalis, T.glabrata) and four bacterial strains (E.coli, S.aureus, K.pneumonie) was performed. This resulted in a single product of 1364 bp with A.fumigatus DNA, while no such product was found in PCR's of other species. Control hybridisation with an A.fumigatus DNA probe confirmed these results. Conclusion : the use of RAPD R5 primer offers a fast and rather easy PCR technique identification of A.fumigatus as well as a possibility to discriminate it from most other fungi and yeasts. Its potential use in analysis of different clinical specimens for confirming the presence of A.fumigatus DNA is currently under progress.
Full conference title:
The 2nd Meeting of the European Confederation of Medical Mycology
- ECMM 2nd (1995)