A microtiter plate-based quantitative method to monitor the growth rate of dermatophytes and test antifungal activity.

Dermatophytosis is one of the most common superficial fungal infections, which is mainly caused by filamentous fungi such as Trichophyton species. A challenging aspect in dermatophyte research is the lack of a straightforward method to measure the rate of growth, in particular when growing dermatophytes in small volumes such as in microtitre plates. However, one characteristic of dermatophytes is their ability to produce compounds such as ammonia that make the growth medium more alkaline. The objective of this study was to test whether the change in pH in a liquid medium, colourimetrically established using the indicator phenol red, was linearly and directly proportional to the growth rate for Trichophyton rubrum and Trichophyton interdigitale. The changes in the colour determined by the phenol-red based assay showed a good correlation with the amount of fungal biomass over an incubation period of 24-120 h. The functionality of the phenol red assay was also validated in experiments on the growth of T. rubrum in the presence of antifungals. The changes in colour showed a clear dose-response relationship compounds and enabled determination of the minimum inhibitory concentration. The phenol red assay is thus a simple and straightforward assay to monitor the rate of growth of Trichophyton spp. and test antifungal activity.