Application of a One-Step method for rapid detection of nucleic acids from fungi.

PCR-based techniques play a crucial role in genotyping and genetic screening in fungal biology. Rapid access to nucleic acids for these reactions can significantly improve the efficiency of fungal analysis, especially when multiple samples need to be tested. In this study, we introduced a simple and rapid method for detecting small amounts of fungal DNA or RNA, named the One-Step method, and confirmed its applicability across various experimental scenarios for fungal detection. The method involves scraping a small quantity of spores or mycelium into sterile water, followed by heat shock, vortexing, and centrifugation to obtain a supernatant that serves as a template for the PCR reaction. Notably, nucleic acids were successfully extracted using the One-Step method from four different genera of fungi-Neurospora crassa, Aspergillus fumigatus, Fusarium oxysporum, and Schizosaccharomyces pombe, as well as from two mycovirus-containing strains, yielding reliable results in PCR identification. However, the nucleic acids of Cryptococcus neoformans were not successfully extracted using the One-Step method, possibly due to the challenge of cleaving its polysaccharide capsule. Taken together, the One-Step method significantly reduces nucleic acids extraction time while enhancing strain screening efficiency in four different fungi, indicating a broad applicability in fungal biology.