title: The use of next-generation sequencing in clinical diagnosis of familial hypercholesterolemia.
abstract: OBJECTIVE
  Familial hypercholesterolemia is a common Mendelian disorder associated with early-onset coronary heart disease that can be treated by cholesterol-lowering drugs. The majority of cases in the United Kingdom are currently without a molecular diagnosis, which is partly due to the cost and time associated with standard screening techniques. The main purpose of this study was to test the sensitivity and specificity of two next-generation sequencing protocols for genetic diagnosis of familial hypercholesterolemia.
  
  
  METHODS
  Libraries were prepared for next-generation sequencing by two target enrichment protocols; one using the SureSelect Target Enrichment System and the other using the PCR-based Access Array platform.
  
  
  RESULTS
  In the validation cohort, both protocols showed 100% specificity, whereas the sensitivity for short variant detection was 100% for the SureSelect Target Enrichment and 98% for the Access Array protocol. Large deletions/duplications were only detected using the SureSelect Target Enrichment protocol. In the prospective cohort, the mutation detection rate using the Access Array was highest in patients with clinically definite familial hypercholesterolemia (67%), followed by patients with possible familial hypercholesterolemia (26%).
  
  
  CONCLUSIONS
  We have shown the potential of target enrichment methods combined with next-generation sequencing for molecular diagnosis of familial hypercholesterolemia. Adopting these assays for patients with suspected familial hypercholesterolemia could improve cost-effectiveness and increase the overall number of patients with a molecular diagnosis.
authors.affiliation: MRC Clinical Sciences Centre, Faculty of Medicine, Imperial College London, London, UK.
authors.forename: Jana
authors.initials: J
authors.lastname: Vandrovcova
authors.forename: Ellen R A
authors.initials: ER
authors.lastname: Thomas
authors.forename: Santosh S
authors.initials: SS
authors.lastname: Atanur
authors.forename: Penny J
authors.initials: PJ
authors.lastname: Norsworthy
authors.forename: Clare
authors.initials: C
authors.lastname: Neuwirth
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Tan
authors.forename: Dalia
authors.initials: D
authors.lastname: Kasperaviciute
authors.forename: Jennifer
authors.initials: J
authors.lastname: Biggs
authors.forename: Laurence
authors.initials: L
authors.lastname: Game
authors.forename: Michael
authors.initials: M
authors.lastname: Mueller
authors.forename: Anne K
authors.initials: AK
authors.lastname: Soutar
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Aitman
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D002648:Child; D002675:Child, Preschool; D056915:DNA Copy Number Variations; D059014:High-Throughput Nucleotide Sequencing / Q000191:economics; D006801:Humans; D006938:Hyperlipoproteinemia Type II / Q000175:diagnosis* / Q000235:genetics; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques; D059645:Mutation Rate; D011446:Prospective Studies; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D055815:Young Adult