Brief communication Investigation of next-generation sequencing technologies as a diagnostic tool for amyotrophic lateral sclerosis Sarah Morgan a, b , Maryam Shoai a, b , Pietro Fratta c , Katie Sidle d , Richard Orrell d , Mary G. Sweeney e , Aleksey Shatunov f , William Sproviero f , Ashley Jones f , Ammar Al-Chalabi f , Andrea Malaspina g , Henry Houlden b , John Hardy a, b , Alan Pittman a, b, * a Reta Lila Weston Institute, UCL Institute of Neurology, London, UK b Department of Molecular Neuroscience, UCL Institute of Neurology, London, UK c Department of Neurodegenerative Diseases, UCL Institute of Neurology, London, UK d Department of Clinical Neuroscience, UCL Institute of Neurology, London, UK e Neurogenetics Unit, National Hospital for Neurology and Neurosurgery, London, UK f Department of Clinical Neuroscience, King’s College London, Institute of Psychiatry, London, UK g Centre for Neuroscience and Trauma, Blizard Institute, Queen Mary University of London, North-East London and Essex Regional Motor Neuron Disease Care Centre, London, UK article info Article history: Received 12 November 2014 Received in revised form 12 December 2014 Accepted 13 December 2014 Available online 20 December 2014 Keywords: Amyotrophic lateral sclerosis ALS Diagnosis Genetic Sequencing Next-generation sequencing NGS MiSeq Neurogenetics abstract The future of genetic diagnostics will see a move toward massively parallel next-generation sequencing of a patient’s DNA. Amyotrophic lateral sclerosis (ALS) is one of the diseases that would benefit from this prospect. Exploring this idea, we designed a screening panel to sequence 25 ALS-linked genes and examined samples from 95 patients with both familial and sporadic ALS. Forty-three rare polymorphisms were detected in this cohort. A third of these have already been reported with respect to ALS, leaving 28 novel variants all open for further investigation. This study highlights the potential benefits of next- generation sequencing as a reliable, cost and time efficient, diagnostic, and research tool for ALS. Ó 2015 Elsevier Inc. All rights reserved. 1. Introduction Rapid diagnosis of amyotrophic lateral sclerosis (ALS) and other neurologic disorders is vital if future treatments are to be applied at an early disease stage. For genetic causes of these diseases, the current technology lies with sequential Sanger sequencing. How- ever, with an array of multiple genes causing each disease and, additionally, numerous alterations within each gene being poten- tially harmful, it can be time consuming and costly to diagnose a patient suspected of harboring a detrimental genetic variation. Furthermore, the range of genetic tests at each institution can be limited. It is now plausible that next-generation sequencing (NGS) technologies will eliminate many of these issues. To test this pos- sibility, we have developed a single comprehensive assay contain- ing 25 genes which have, to varying degrees, been implicated in ALS. 2. Methods 2.1. Probe design The ALS gene panel was designed using Illumina TruSeq Custom Amplicon and implemented on an Illumina MiSeq platform. This utilizes polymerase chain reaction amplicon-based target enrich- ment and screens for variants in 25 ALS disease genes. These were split into 2 groups depending on the desired coverage. The first * Corresponding author at: Department of Molecular Neuroscience, UCL Institute of Neurology, 9th Floor, Queen Square, London WC1N 3BG, UK. Tel.: þ44 207 692 3076; fax: þ44 203 448 4017. E-mail address: a.pittman@ucl.ac.uk (A. Pittman). Contents lists available at ScienceDirect Neurobiology of Aging journal homepage: www.elsevier.com/locate/neuaging 0197-4580/$ e see front matter Ó 2015 Elsevier Inc. All rights reserved. http://dx.doi.org/10.1016/j.neurobiolaging.2014.12.017 Neurobiology of Aging 36 (2015) 1600.e5e1600.e8