[Molecular genetic methods in medical mycology: current state and perspectives]

Abstract

Molecular genetic methods are becoming increasingly important as a part of diagnostics in the clinical mycology laboratory. The methods may be used to detect nucleic acids of fungi in clinical samples, to identify fungal cultures to the species level or to evaluate inter-strain differences within the species. For these purposes, numerous techniques have been developed over the past years. Recently, the effort to accept some of them as a widely accepted standard has increased, particularly in rapid diagnosis of invasive aspergillosis. Various modifications of the PCR are used to detect DNA in clinical material, particularly nested and real-time PCR. Restriction endonuclease analysis, usually followed by Southern blot hybridization, as well as karyotyping by pulsed-field gel electrophoresis are often used as typing methods. In addition, a group of PCR-based typing techniques is developed extensively. These include random amplified polymorphic DNA, amplified fragment length polymorphism, inter-repeat PCR, p olymorphic microsatellite markers, amplification with sequence-specific primers, multilocus sequence typing and DNA microarray technique. The latter methods are considered to be most likely accepted as the gold standard in the future.