Masvidal L.,Human Molecular Genetics Group |
Alvarez A.,Cystic Fibrosis Adult Unit |
Ruano L.,Cystic Fibrosis Adult Unit |
de Gracia J.,Cystic Fibrosis Adult Unit |
And 2 more authors.
Journal of Cystic Fibrosis | Year: 2012
Background: CFTR expression studies contribute in understanding the relationship between CFTR transcripts and clinical outcomes. Normalization of qPCR data is an essential step to determine target gene expression. Consequently, appropriate reference genes must be selected for each gene/tissue. In this work, we have assessed the suitability of four potential reference genes for CFTR expression analysis in nasal epithelium. Methods: B2M, GUSB, HPRT1 and ATP2B4 expression was evaluated in nasal epithelium samples (CFTR-wt controls, n. =21; CFTR-splicing group, n. =18) by RT-qPCR. Calibration curves were built and different analyses (geNorm, NormFinder, Mann-Whitney) were performed to evaluate gene expression stability between samples as well as between groups. Results and conclusions: We have applied an accurate approach to select reference genes for CFTR expression analysis in nasal epithelium. From the four genes assessed, GUSB and ATP2B4 have been validated as a reliable gene combination for CFTR gene qPCR data normalization. © 2012 European Cystic Fibrosis Society.. Source