Get access

Genetic diagnosis of familial breast cancer using clonal sequencing

Authors

  • Joanne E. Morgan,

    1. University of Leeds Institute of Molecular Medicine, Wellcome Trust Brenner Building, St. James's University Hospital, Leeds, United Kingdom
    Search for more papers by this author
  • Ian M. Carr,

    1. University of Leeds Institute of Molecular Medicine, Wellcome Trust Brenner Building, St. James's University Hospital, Leeds, United Kingdom
    Search for more papers by this author
  • Eamonn Sheridan,

    1. University of Leeds Institute of Molecular Medicine, Wellcome Trust Brenner Building, St. James's University Hospital, Leeds, United Kingdom
    Search for more papers by this author
  • Carol E. Chu,

    1. Department of Clinical Genetics, Leeds Teaching Hospitals Trust St. James's University Hospital, Leeds, United Kingdom
    Search for more papers by this author
  • Bruce Hayward,

    1. University of Leeds Institute of Molecular Medicine, Wellcome Trust Brenner Building, St. James's University Hospital, Leeds, United Kingdom
    Search for more papers by this author
  • Nick Camm,

    1. Department of Clinical Genetics, Leeds Teaching Hospitals Trust St. James's University Hospital, Leeds, United Kingdom
    Search for more papers by this author
  • Helen A. Lindsay,

    1. Department of Clinical Genetics, Leeds Teaching Hospitals Trust St. James's University Hospital, Leeds, United Kingdom
    Search for more papers by this author
  • Chris J. Mattocks,

    1. National Genetics Reference Laboratory (Wessex), Salisbury District Hospital, Salisbury, United Kingdom
    Search for more papers by this author
  • Alexander F. Markham,

    1. University of Leeds Institute of Molecular Medicine, Wellcome Trust Brenner Building, St. James's University Hospital, Leeds, United Kingdom
    Search for more papers by this author
  • David T. Bonthron,

    1. University of Leeds Institute of Molecular Medicine, Wellcome Trust Brenner Building, St. James's University Hospital, Leeds, United Kingdom
    Search for more papers by this author
  • Graham R. Taylor

    Corresponding author
    1. University of Leeds Institute of Molecular Medicine, Wellcome Trust Brenner Building, St. James's University Hospital, Leeds, United Kingdom
    2. Department of Clinical Genetics, Leeds Teaching Hospitals Trust St. James's University Hospital, Leeds, United Kingdom
    • Leeds Institute of Molecular Medicine, Experimental Therapeutics, 6.2 Clinical Sciences Building, St. James's University Hospital, Beckett Street, Leeds LS9 7TF, UK
    Search for more papers by this author

  • Communicated by Arupa Ganguly

Abstract

Using conventional Sanger sequencing as a reference standard, we compared the sensitivity, specificity, and capacity of the Illumina GA II platform for the detection of TP53, BRCA1, and BRCA2 mutations in established tumor cell lines and DNA from patients with germline mutations. A total of 656 coding variants were identified in four cell lines and 65 patient DNAs. All of the known pathogenic mutations (including point mutations and insertions/deletions of up to 16 nucleotides) were identified, using a combination of the Illumina data analysis pipeline with custom and commercial sequence alignment software. In our configuration, clonal sequencing outperforms current diagnostic methods, providing a reduction in analysis times and in reagent costs compared with conventional sequencing. These improvements open the possibility of BRCA1/2 testing for a wider spectrum of at-risk women, and will allow the genetic classification of tumors prior to the use of novel PARP inhibitors to treat BRCA-deficient breast cancers. Hum Mutat 31:1–8, 2010. © 2010 Wiley-Liss, Inc.

Get access to the full text of this article

Ancillary