New Reference Genes for qRT-PCR Analysis as a Potential Target for Identification of <i>Trichophyton verrucosum</i> in Different Culture Conditions

Dermatophytes are a group of filamentous fungi infecting skin, hair, and nails that raise great diagnostic difficulties. qRT-PCR is a reliable technique for quantifying gene expression with increasingly frequent use in mycological diagnostics. Knowledge of genes and molecular markers with potential...

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Bibliographic Details
Main Authors: Sebastian Gnat, Dominik Łagowski, Aneta Nowakiewicz, Aleksandra Trościańczyk, Mariusz Dyląg
Format: article
Language:EN
Published: MDPI AG 2021
Subjects:
R
Online Access:https://doaj.org/article/35aa5e9e8b2f4008b4f4f7c67f6930f4
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Summary:Dermatophytes are a group of filamentous fungi infecting skin, hair, and nails that raise great diagnostic difficulties. qRT-PCR is a reliable technique for quantifying gene expression with increasingly frequent use in mycological diagnostics. Knowledge of genes and molecular markers with potential to be used in the identification of dermatophytes is of great importance for the development of this branch of diagnostics. In this article, the suitability of six candidate reference genes (<i>TUBB</i>, <i>ACTB</i>, <i>ADPRF</i>, <i>RPL2</i>, <i>SDHA</i>, and <i>EEF1A1</i>) was investigated for gene expression analysis in the dermatophyte <i>Trichophyton verrucosum</i>, which was cultured in various mycological media that are commonly used in a diagnostic laboratory, i.e., Sabouraud, potato dextrose, and keratin-supplemented MM-Cove. The different culture conditions are extremely important factors for the growth and physiology of dermatophytes. Gene expression stability was evaluated using geNorm, NormFinder, BestKeeper, and RefFinder algorithms. Regarding the stability of expression, <i>SDHA</i> was the most stable housekeeping gene; hence, this gene is recommended for future qRT-PCR studies on <i>T. verrucosum</i> strains. These results allow us to conclude that the <i>SDHA</i> gene can be an additional good candidate as an identification target in the qRT-PCR technique.