title: rpb2 is a reliable reference gene for quantitative gene expression analysis in the dermatophyte Trichophyton rubrum.
abstract: The selection of reference genes used for data normalization to quantify gene expression by real-time PCR amplifications (qRT-PCR) is crucial for the accuracy of this technique. In spite of this, little information regarding such genes for qRT-PCR is available for gene expression analyses in pathogenic fungi. Thus, we investigated the suitability of eight candidate reference genes in isolates of the human dermatophyte Trichophyton rubrum subjected to several environmental challenges, such as drug exposure, interaction with human nail and skin, and heat stress. The stability of these genes was determined by geNorm, NormFinder and Best-Keeper programs. The gene with the most stable expression in the majority of the conditions tested was rpb2 (DNA-dependent RNA polymerase II), which was validated in three T. rubrum strains. Moreover, the combination of rpb2 and chs1 (chitin synthase) genes provided for the most reliable qRT-PCR data normalization in T. rubrum under a broad range of biological conditions. To the best of our knowledge this is the first report on the selection of reference genes for qRT-PCR data normalization in dermatophytes and the results of these studies should permit further analysis of gene expression under several experimental conditions, with improved accuracy and reliability.
authors.affiliation: Departamento de Genética, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, São Paulo, Brazil.
authors.forename: Tiago R
authors.initials: TR
authors.lastname: Jacob
authors.forename: Nalu T A
authors.initials: NT
authors.lastname: Peres
authors.forename: Gabriela F
authors.initials: GF
authors.lastname: Persinoti
authors.forename: Larissa G
authors.initials: LG
authors.lastname: Silva
authors.forename: Mendelson
authors.initials: M
authors.lastname: Mazucato
authors.forename: Antonio
authors.initials: A
authors.lastname: Rossi
authors.forename: Nilce M
authors.initials: NM
authors.lastname: Martinez-Rossi
mesh_terms: D005656:Fungal Proteins / Q000235:genetics*; D020869:Gene Expression Profiling / Q000592:standards*; D015966:Gene Expression Regulation, Fungal; D006801:Humans; D012319:RNA Polymerase II / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012015:Reference Standards; D014249:Trichophyton / Q000235:genetics*
chemical_list: D005656:Fungal Proteins; D012319:RNA Polymerase II