title: Next-generation sequencing for disorders of low and high bone mineral density.
abstract: To achieve an efficient molecular diagnosis of osteogenesis imperfecta (OI), Ehlers-Danlos syndrome (EDS), and osteopetrosis (OPT), we designed a next-generation sequencing (NGS) platform to sequence 34 genes. We validated this platform on known cases and have successfully identified the causative mutation in most patients without a prior molecular diagnosis.
  
  
  BACKGROUND
  Osteogenesis imperfecta, Ehlers-Danlos syndrome, and osteopetrosis are collectively common inherited skeletal diseases. Evaluation of subjects with these conditions often includes molecular testing which has important counseling and therapeutic and sometimes legal implications. Since several different genes have been implicated in these conditions, Sanger sequencing of each gene can be a prohibitively expensive and time-consuming way to reach a molecular diagnosis.
  
  
  METHODS
  In order to circumvent these problems, we have designed and tested a NGS platform that would allow simultaneous sequencing on a single diagnostic platform of different genes implicated in OI, OPT, EDS, and other inherited conditions, leading to low or high bone mineral density. We used a liquid-phase probe library that captures 602 exons (~100 kb) of 34 selected genes and have applied it to test clinical samples from patients with bone disorders.
  
  
  RESULTS
  NGS of the captured exons by Illumina HiSeq 2000 resulted in an average coverage of over 900X. The platform was successfully validated by identifying mutations in six patients with known mutations. Moreover, in four patients with OI or OPT without a prior molecular diagnosis, the assay was able to detect the causative mutations.
  
  
  CONCLUSIONS
  In conclusion, our NGS panel provides a fast and accurate method to arrive at a molecular diagnosis in most patients with inherited high or low bone mineral density disorders.
authors: Department of Molecular and Human Genetics, Baylor College of Medicine, One Baylor Plaza, R814, MS225, Houston, TX 77030, USA.
authors: G
authors: G
authors: Sule
authors: P M
authors: PM
authors: Campeau
authors: V W
authors: VW
authors: Zhang
authors: S C S
authors: SC
authors: Nagamani
authors: B C
authors: BC
authors: Dawson
authors: M
authors: M
authors: Grover
authors: C A
authors: CA
authors: Bacino
authors: V R
authors: VR
authors: Sutton
authors: N
authors: N
authors: Brunetti-Pierri
authors: J T
authors: JT
authors: Lu
authors: E
authors: E
authors: Lemire
authors: R A
authors: RA
authors: Gibbs
authors: D H
authors: DH
authors: Cohn
authors: H
authors: H
authors: Cui
authors: L-J
authors: LJ
authors: Wong
authors: B H
authors: BH
authors: Lee
mesh_terms: D000328:Adult; D015519:Bone Density / Q000235:genetics*; D001848:Bone Diseases, Developmental / Q000175:diagnosis* / Q000235:genetics* / Q000503:physiopathology; D004535:Ehlers-Danlos Syndrome / Q000175:diagnosis / Q000235:genetics / Q000503:physiopathology; D015723:Gene Library; D020022:Genetic Predisposition to Disease; D005820:Genetic Testing / Q000379:methods; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D008297:Male; D009154:Mutation; D010013:Osteogenesis Imperfecta / Q000175:diagnosis / Q000235:genetics / Q000503:physiopathology; D010022:Osteopetrosis / Q000175:diagnosis / Q000235:genetics / Q000503:physiopathology; D017422:Sequence Analysis, DNA / Q000379:methods