BRCA Lab

The BRCA Lab at Department of Oncology, Lund University, provides BRCA1 and BRCA2 mutations screening and predictive testing as a clinical service for oncogenetic clinics in Sweden (Lund, Stockholm, Uppsala, Göteborg, Linköping and Umeå) in their counseling of breast and breast-ovarian cancer families. Current technologies make use of Illumina Genome Analyzer single-read sequencing of barcoded and captured genomic fragments of BRCA1 and BRCA2 from blood-DNA, for detection of germline base substitutions, deletions and insertions in both coding and noncoding/nonrepetitive regions. This sequencing is complemented with multiplex-ligation dependent probe amplification (MLPA) analysis for detection of larger genomic deletions and duplications. For novel rearrangements detected by MLPA, a zoom-in microarray comparative genomic hybridization analysis is used to delineate the breakpoints for design of mutation specific PCR primers. Sanger sequencing or pyrosequencing is used for analysis of known single mutations (substitutions and small deletions or insertions) in predictive tests. Screening for multiple known BRCA gene mutations (>200 distinct BRCA1 or BRCA2 mutations have been identified in Swedish breast cancer families) is performed by Sequenom MassARRAY technology, making analysis also of formalin-fixed paraffin-embedded tissue possible. The Illumina Genome Analyzer BRCA gene screening will in 2011 include also other breast cancer susceptibility genes by using an extended capture design and paired-end sequencing.