Advances in Alport syndrome diagnosis using next generation sequencing

Alessandra Renieri (1) , Rosangela Artuso (1) , Chiara Fallerini (1) , Laura Dosa (1) , Francesca Scionti (1) , Maurizio Clementi (1) , Guido Garosi (1) , Laura Massella (1) , Maria Carmela Epistolato (1) , Roberta Mancini (1) , Francesca Mari (1) , Ilaria Longo (1) , Francesca Ariani (1) , Mirella Bruttini (1)

1 Biotechnology

Alport syndrome is a hereditary nephropathy often associated with sensorineural hypoacusis and ocular abnormalities. Mutations in the COL4A5 gene cause X-linked Alport syndrome. Mutations in COL4A4 and COL4A3 genes have been reported in both autosomal recessive and autosomal dominant Alport syndrome. The conventional mutation screening, performed by DHPLC and/or Sanger sequencing, is time consuming and has relatively high costs due to the absence of hot spots and to the high number of exons per gene: 51 (COL4A5), 48 (COL4A4), and 52 (COL4A3). Several months are usually necessary to complete the diagnosis, especially in cases with less informative pedigrees. To overcome these limitations, we designed a next generation sequencing protocol enabling simultaneous detection of all possible variants in the three genes. We used a method coupling selective amplification to the 454 Roche DNA-sequencing platform (GS junior). The application of this technology allowed us to identify the second mutation in two Alport syndrome patients (p.Ser1147Phe in COL4A3 and p.Arg1682Trp in COL4A4) and to reconsider the diagnosis of Alport syndrome in a third patient. This study, therefore, illustrates the successful application of next generation sequencing to mutation screening of Mendelian disorders with locus heterogeneity.

Mots clés: DNA sequencing; next generation sequencing; Alport syndrome

Domaines: Génétique

Alessandra Renieri, Rosangela Artuso, Chiara Fallerini, Laura Dosa, Francesca Scionti, et al.. Advances in Alport syndrome diagnosis using next generation sequencing. European Journal of Human Genetics, 2011, ⟨10.1038/ejhg.2011.164⟩. ⟨hal-00677002⟩