RAPD-PCR for Typing and Identification of Aspergillus fumigatus

RAPD is a PCR variant that uses a single primer and is performed based on the principle that, as the annealing temperature decreases, the primer finds multiple sequences in the template DNA with which it can hybridize when these sites are close to each other and in opposite orientations, resulting in amplification of the DNA sequence between the sites to produce a template DNA band. The RAPD method has been used for the typing and identification of Aspergillus species, characterization of environmental sources, and the detection of intraspecific and interspecific genetic variability. For this purpose, a wide variety of primers have been used with varying results. Primer OPF-01 has proven helpful in typing Aspergillus spp. and identifying A. fumigatus, as it generates species-specific polymorphic patterns for this fungus. This chapter describes the RAPD method for identifying A. fumigatus with primer OPF-01.