BACKGROUND: Comprehensive resequencing of the causative and disease-related genes of neurodegenerative diseases is expected to enable (1) comprehensive mutational analysis of familial cases, (2) identification of sporadic cases with de novo or low-penetrant mutations, (3) identification of rare variants conferring disease susceptibility, and ultimately (4) better understanding of the molecular basis of these diseases. OBJECTIVE: To develop a microarray-based high-throughput resequencing system for the causative and disease-related genes of amyotrophic lateral sclerosis (ALS) and other neurodegenerative diseases. DESIGN: Validation of the system was conducted in terms of the signal-to-noise ratio, accuracy, and throughput. Comprehensive gene analysis was applied for patients with ALS. Subjects Ten patients with familial ALS, 35 patients with sporadic ALS, and 238 controls. RESULTS: The system detected point mutations with 100% accuracy and completed the resequencing of 270 kilobase pairs in 3 working days with greater than 99.9% accuracy of base calls, or the determination of base(s) at each position. Analysis of patients with familial ALS revealed 2 SOD1 mutations. Analysis of the 35 patients with sporadic ALS revealed a previously known SOD1 mutation, S134N, a novel putative pathogenic DCTN1 mutation, R997W, and 9 novel variants including 4 nonsynonymous heterozygous variants consisting of 2 in ALS2, 1 in ANG, and 1 in VEGF that were not found in the controls. CONCLUSION: The DNA microarray-based resequencing system is a powerful tool for high-throughput comprehensive analysis of causative and disease-related genes. It can be used to detect mutations in familial and sporadic cases and to identify numerous novel variants potentially associated with genetic risks.
BACKGROUND: Comprehensive resequencing of the causative and disease-related genes of neurodegenerative diseases is expected to enable (1) comprehensive mutational analysis of familial cases, (2) identification of sporadic cases with de novo or low-penetrant mutations, (3) identification of rare variants conferring disease susceptibility, and ultimately (4) better understanding of the molecular basis of these diseases. OBJECTIVE: To develop a microarray-based high-throughput resequencing system for the causative and disease-related genes of amyotrophic lateral sclerosis (ALS) and other neurodegenerative diseases. DESIGN: Validation of the system was conducted in terms of the signal-to-noise ratio, accuracy, and throughput. Comprehensive gene analysis was applied for patients with ALS. Subjects Ten patients with familial ALS, 35 patients with sporadic ALS, and 238 controls. RESULTS: The system detected point mutations with 100% accuracy and completed the resequencing of 270 kilobase pairs in 3 working days with greater than 99.9% accuracy of base calls, or the determination of base(s) at each position. Analysis of patients with familial ALS revealed 2 SOD1 mutations. Analysis of the 35 patients with sporadic ALS revealed a previously known SOD1 mutation, S134N, a novel putative pathogenic DCTN1 mutation, R997W, and 9 novel variants including 4 nonsynonymous heterozygous variants consisting of 2 in ALS2, 1 in ANG, and 1 in VEGF that were not found in the controls. CONCLUSION: The DNA microarray-based resequencing system is a powerful tool for high-throughput comprehensive analysis of causative and disease-related genes. It can be used to detect mutations in familial and sporadic cases and to identify numerous novel variants potentially associated with genetic risks.
Authors: Yuji Takahashi; Yoko Fukuda; Jun Yoshimura; Atsushi Toyoda; Kari Kurppa; Hiroyoko Moritoyo; Veronique V Belzil; Patrick A Dion; Koichiro Higasa; Koichiro Doi; Hiroyuki Ishiura; Jun Mitsui; Hidetoshi Date; Budrul Ahsan; Takashi Matsukawa; Yaeko Ichikawa; Takashi Moritoyo; Mayumi Ikoma; Tsukasa Hashimoto; Fumiharu Kimura; Shigeo Murayama; Osamu Onodera; Masatoyo Nishizawa; Mari Yoshida; Naoki Atsuta; Gen Sobue; Jennifer A Fifita; Kelly L Williams; Ian P Blair; Garth A Nicholson; Paloma Gonzalez-Perez; Robert H Brown; Masahiro Nomoto; Klaus Elenius; Guy A Rouleau; Asao Fujiyama; Shinichi Morishita; Jun Goto; Shoji Tsuji Journal: Am J Hum Genet Date: 2013-10-10 Impact factor: 11.025
Authors: Takuya Konno; Owen A Ross; Hélio A G Teive; Jarosław Sławek; Dennis W Dickson; Zbigniew K Wszolek Journal: Parkinsonism Relat Disord Date: 2017-06-12 Impact factor: 4.891
Authors: Hong-Ying Wang; Vivek Gopalan; Ivona Aksentijevich; Meredith Yeager; Chi Adrian Ma; Yasmin Ali Mohamoud; Mariam Quinones; Casey Matthews; Joseph Boland; Julie E Niemela; Troy R Torgerson; Silvia Giliani; Gulbu Uzel; Jordan S Orange; Ralph Shapiro; Luigi Notarangelo; Hans D Ochs; Thomas Fleisher; Daniel Kastner; Stephen J Chanock; Ashish Jain Journal: Hum Mutat Date: 2010-09 Impact factor: 4.878
Authors: Jin Song; Nizar Smaoui; Radha Ayyagari; David Stiles; Sonia Benhamed; Ian M MacDonald; Stephen P Daiger; Santa J Tumminia; Fielding Hejtmancik; Xinjing Wang Journal: Invest Ophthalmol Vis Sci Date: 2011-11-25 Impact factor: 4.799