title: Comprehensive molecular diagnosis of 179 Leber congenital amaurosis and juvenile retinitis pigmentosa patients by targeted next generation sequencing.
abstract: BACKGROUND
  Leber congenital amaurosis (LCA) and juvenile retinitis pigmentosa (RP) are inherited retinal diseases that cause early onset severe visual impairment. An accurate molecular diagnosis can refine the clinical diagnosis and allow gene specific treatments.
  
  
  METHODS
  We developed a capture panel that enriches the exonic DNA of 163 known retinal disease genes. Using this panel, we performed targeted next generation sequencing (NGS) for a large cohort of 179 unrelated and prescreened patients with the clinical diagnosis of LCA or juvenile RP. Systematic NGS data analysis, Sanger sequencing validation, and segregation analysis were utilised to identify the pathogenic mutations. Patients were revisited to examine the potential phenotypic ambiguity at the time of initial diagnosis.
  
  
  RESULTS
  Pathogenic mutations for 72 patients (40%) were identified, including 45 novel mutations. Of these 72 patients, 58 carried mutations in known LCA or juvenile RP genes and exhibited corresponding phenotypes, while 14 carried mutations in retinal disease genes that were not consistent with their initial clinical diagnosis. We revisited patients in the latter case and found that homozygous mutations in PRPH2 can cause LCA/juvenile RP. Guided by the molecular diagnosis, we reclassified the clinical diagnosis in two patients.
  
  
  CONCLUSIONS
  We have identified a novel gene and a large number of novel mutations that are associated with LCA/juvenile RP. Our results highlight the importance of molecular diagnosis as an integral part of clinical diagnosis.
authors.affiliation: Human Genome Sequencing Center, Baylor College of Medicine, Houston, Texas, USA.
authors.forename: Xia
authors.initials: X
authors.lastname: Wang
authors.forename: Hui
authors.initials: H
authors.lastname: Wang
authors.forename: Vincent
authors.initials: V
authors.lastname: Sun
authors.forename: Han-Fang
authors.initials: HF
authors.lastname: Tuan
authors.forename: Vafa
authors.initials: V
authors.lastname: Keser
authors.forename: Keqing
authors.initials: K
authors.lastname: Wang
authors.forename: Huanan
authors.initials: H
authors.lastname: Ren
authors.forename: Irma
authors.initials: I
authors.lastname: Lopez
authors.forename: Jacques E
authors.initials: JE
authors.lastname: Zaneveld
authors.forename: Sorath
authors.initials: S
authors.lastname: Siddiqui
authors.forename: Stephanie
authors.initials: S
authors.lastname: Bowles
authors.forename: Ayesha
authors.initials: A
authors.lastname: Khan
authors.forename: Jason
authors.initials: J
authors.lastname: Salvo
authors.forename: Samuel G
authors.initials: SG
authors.lastname: Jacobson
authors.forename: Alessandro
authors.initials: A
authors.lastname: Iannaccone
authors.forename: Feng
authors.initials: F
authors.lastname: Wang
authors.forename: David
authors.initials: D
authors.lastname: Birch
authors.forename: John R
authors.initials: JR
authors.lastname: Heckenlively
authors.forename: Gerald A
authors.initials: GA
authors.lastname: Fishman
authors.forename: Elias I
authors.initials: EI
authors.lastname: Traboulsi
authors.forename: Yumei
authors.initials: Y
authors.lastname: Li
authors.forename: Dianna
authors.initials: D
authors.lastname: Wheaton
authors.forename: Robert K
authors.initials: RK
authors.lastname: Koenekoop
authors.forename: Rui
authors.initials: R
authors.lastname: Chen
mesh_terms: D000483:Alleles; D000595:Amino Acid Sequence; D001483:Base Sequence; D059472:Exome; D005260:Female; D005838:Genotype; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D057130:Leber Congenital Amaurosis / Q000175:diagnosis* / Q000235:genetics; D009154:Mutation; D010375:Pedigree; D020641:Polymorphism, Single Nucleotide; D015203:Reproducibility of Results; D012174:Retinitis Pigmentosa / Q000175:diagnosis* / Q000235:genetics; D012680:Sensitivity and Specificity
keywords: Clinical genetics; Genetic screening/counselling; Ophthalmology; Vision research