Actin Gene-Targeted RT-PCR Could be a Useful Method for Evaluating In Vitro Fungicidal Activity against Dermatophytes

Abstract

This study examined the effects of the antifungal agents amorolfine hydrochloride (AMF) and bifonazole (BFZ) on actin mRNA expression determined by reverse transcription-polymerase chain reaction (RT-PCR) amplification, and the morphology of Trichophyton mentagrophytes. In AMF-treated cultures, the hyphal cells of T. mentagrophytes exhibited concentration- and/or time-dependent progressively degenerative morphological damage. Those cultures exhibiting severe and necrotic morphological changes (such as crushed, bent and flattened hyphal cells) did not express actin mRNA. In BFZ-treated cultures, similar morphological changes were seen in the hyphae of T. mentagrophytes, but these changes were milder than observed in the AMF-treated samples, and actin gene fragments were amplified in all of these samples. These results indicate that actin gene-targeted RT-PCR could be a useful tool for testing the susceptibility of dermatophytes to antifungal agents in vitro.