Typing of Aspergillus Fumigatus and Aspergillus Niger Strains by Random Amplification of Polymorphic DNA Analysis Using a Six Primer Set.

Abstract:

Differentiating and typing clinical and environmental isolates of Aspergillus strains are necessary for epidemiological studies and could contribute to the solution of several pertinent clinical problems. We investigated the utility of the RAPD-PCR technique for typing twenty-six strains of Aspergillus fumigatus and ten additional clinical and environmental Aspergillus fumigatus and Aspergillus niger isolates. Fungal genomic DNA was extracted using glass bead disruption and RAPD analysis was performed by using a six individual (R108, R151, UBC90) and combined (R108+R151, R108+UBS90, R151+UBC90) primer set. Primer pair R108+UBC90 demonstrated the highest discriminatory power with collection strains, whereas primer R151 displayed the highest degree of discrimination power with clinical and environmental isolates. Both primer sets detected seven types of strains. It was found that primer R151, on its own and in combination with primer UBC90, shared a relative relationship between clinical and environmental A. fumigatus isolates. We found that although RAPD-PCR analysis can be applied as a simple, rapid, and very effective method for differentiating Aspergillus strains, selecting the best primers is a critical step to reach the desirable discriminatory power.