Improving molecular diagnosis in epilepsy by a dedicated high-throughput sequencing platform

• Published in: European journal of human genetics : EJHG Vol. 23; no. 3; pp. 354 - 362
• Main Authors: Della Mina, Erika, Ciccone, Roberto, Brustia, Francesca, Bayindir, Baran, Limongelli, Ivan, Vetro, Annalisa, Iascone, Maria, Pezzoli, Laura, Bellazzi, Riccardo, Perotti, Gianfranco, De Giorgis, Valentina, Lunghi, Simona, Coppola, Giangennaro, Orcesi, Simona, Merli, Pietro, Savasta, Salvatore, Veggiotti, Pierangelo, Zuffardi, Orsetta
• Format: Journal Article
• Language: English
• Published: England Nature Publishing Group 01-03-2015
• Subjects: Adolescent; Adult; Case-Control Studies; Child; Child, Preschool; Computational Biology; Diagnosis; Epilepsy; Epilepsy - diagnosis; Epilepsy - genetics; Families & family life; Female; Follow-Up Studies; Genes; Genetic Association Studies; Genetic Testing; Genetics; Genomes; Genotype & phenotype; High-Throughput Nucleotide Sequencing; Humans; Infant; Male; Mutation; Patients; Phenotypes; Workflow; Young Adult; Italy
• ISSN: 1018-4813; 1476-5438
• DOI: 10.1038/ejhg.2014.92

We analyzed by next-generation sequencing (NGS) 67 epilepsy genes in 19 patients with different types of either isolated or syndromic epileptic disorders and in 15 controls to investigate whether a quick and cheap molecular diagnosis could be provided. The average number of nonsynonymous and splice site mutations per subject was similar in the two cohorts indicating that, even with relatively small targeted platforms, finding the disease gene is not an univocal process. Our diagnostic yield was 47% with nine cases in which we identified a very likely causative mutation. In most of them no interpretation would have been possible in absence of detailed phenotype and familial information. Seven out of 19 patients had a phenotype suggesting the involvement of a specific gene. Disease-causing mutations were found in six of these cases. Among the remaining patients, we could find a probably causative mutation only in three. None of the genes affected in the latter cases had been suspected a priori. Our protocol requires 8-10 weeks including the investigation of the parents with a cost per patient comparable to sequencing of 1-2 medium-to-large-sized genes by conventional techniques. The platform we used, although providing much less information than whole-exome or whole-genome sequencing, has the advantage that can also be run on 'benchtop' sequencers combining rapid turnaround times with higher manageability.