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Targeted next generation sequencing as a diagnostic tool in epileptic disorders
Author(s) -
Lemke Johannes R.,
Riesch Erik,
Scheurenbrand Tim,
Schubach Max,
Wilhelm Christian,
Steiner Isabelle,
Hansen Jörg,
Courage Carolina,
Gallati Sabina,
Bürki Sarah,
Strozzi Susi,
Simonetti Barbara Goeggel,
Grunt Sebastian,
Steinlin Maja,
Alber Michael,
Wolff Markus,
Klopstock Thomas,
Prott Eva C.,
Lorenz Rüdiger,
Spaich Christiane,
Rona Sabine,
Lakshminarasimhan Maya,
Kröll Judith,
Dorn Thomas,
Krämer Günter,
Synofzik Matthis,
Becker Felicitas,
Weber Yvonne G.,
Lerche Holger,
Böhm Detlef,
Biskup Saskia
Publication year - 2012
Publication title -
epilepsia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.687
H-Index - 191
eISSN - 1528-1167
pISSN - 0013-9580
DOI - 10.1111/j.1528-1167.2012.03516.x
Subject(s) - epilepsy , clinical neurology , medicine , neuroscience , psychology , psychiatry
Summary Purpose: Epilepsies have a highly heterogeneous background with a strong genetic contribution. The variety of unspecific and overlapping syndromic and nonsyndromic phenotypes often hampers a clear clinical diagnosis and prevents straightforward genetic testing. Knowing the genetic basis of a patient’s epilepsy can be valuable not only for diagnosis but also for guiding treatment and estimating recurrence risks. Methods: To overcome these diagnostic restrictions, we composed a panel of genes for Next Generation Sequencing containing the most relevant epilepsy genes and covering the most relevant epilepsy phenotypes known so far. With this method, 265 genes were analyzed per patient in a single step. We evaluated this panel on a pilot cohort of 33 index patients with concise epilepsy phenotypes or with a severe but unspecific seizure disorder covering both sporadic and familial cases. Key Findings: We identified presumed disease‐causing mutations in 16 of 33 patients comprising sequence alterations in frequently as well as in less commonly affected genes. The detected aberrations encompassed known and unknown point mutations ( SCN1A p.R222X, p. E289V, p.379R, p.R393H; SCN2A p.V208E; STXBP1 p.R122X; KCNJ10 p.L68P, p.I129V; KCTD7 p.L108M; KCNQ3 p.P574S; ARHGEF9 p.R290H; SMS p.F58L; TPP1 p.Q278R, p.Q422H; MFSD8 p.T294K), a putative splice site mutation ( SCN1A c.693A> p.T/P231P) and small deletions ( SCN1A p.F1330L fs 3X [1 bp]; MFSD8 p.A138D fs 10X [7 bp]). All mutations have been confirmed by conventional Sanger sequencing and, where possible, validated by parental testing and segregation analysis. In three patients with either Dravet syndrome or myoclonic epilepsy, we detected SCN1A mutations (p.R222X, p.P231P, p.R393H), even though other laboratories had previously excluded aberrations of this gene by Sanger sequencing or high‐resolution melting analysis. Significance: We have developed a fast and cost‐efficient diagnostic screening method to analyze the genetic basis of epilepsies. We were able to detect mutations in patients with clear and with unspecific epilepsy phenotypes, to uncover the genetic basis of many so far unresolved cases with epilepsy including mutation detection in cases in which previous conventional methods yielded falsely negative results. Our approach thus proved to be a powerful diagnostic tool that may contribute to collecting information on both common and unknown epileptic disorders and in delineating associated phenotypes of less frequently mutated genes.