Abstract
Next-generation sequencing (NGS) has enabled new approaches for detection of mutations in the BRCA1 and BRCA2 genes responsible for hereditary breast and ovarian cancer (HBOC). The search for germline mutations in the BRCA1 and BRCA2 genes is of importance with respect to oncogenetic and surgical (bilateral mastectomy, ovariectomy) counselling. Testing tumor material for BRCA mutations is of increasing importance for therapeutic decision making as the poly ADP ribose polymerase (PARP) inhibitor, olaparib, is now available to treat patients with specific forms of ovarian cancer and BRCA mutations. Molecular genetics laboratories should develop reliable and sensitive techniques for the complete analysis of the BRCA1 and BRCA2 genes. This is a challenge due to the size of the coding sequence of the BRCA1/2 genes, the absence of hot spot mutations, and particularly by the lower DNA quality obtained from Formalin-Fixed Paraffin-Embedded (FFPE) tissue. As a result, a number of analyses are uninterpretable and do not always provide a result to the clinician, limiting the optimal therapeutic management of patients. The availability of Fresh Frozen Tissue (FFT) for some laboratories and the excellent quality of the DNA extracted from it offers an alternative. For this reason, we evaluated Multiplicom's BRCA MASTR Dx assay on a set of 97 FFT derived DNA samples, in combination with the MID for Illumina MiSeq for BRCA1 and BRCA2 mutation detection. We obtained interpretable NGS results for all tested samples and showed > 99,7% sensitivity, specificity and accuracy.
Highlights
Generation sequencing (NGS) is used in routine testing for germline mutations that cause rare diseases or hereditary cancers
We showed that Multiplicom’s BRCA MASTR Dx assay provides complete exon coverage of all coding sequences of BRCA1 and BRCA2 genes and combined with Fresh Frozen Tissue (FFT) derived DNA resulted in high sensitivity (≥99.7%), specificity (≥99.99%) and accuracy (≥99.99%)
We showed that FFT derived DNA is superior for diagnostic testing over Formalin-Fixed Paraffin-Embedded (FFPE) derived DNA
Summary
Generation sequencing (NGS) is used in routine testing for germline mutations that cause rare diseases or hereditary cancers. Different laboratories have implemented NGS to analyze BRCA1 and BRCA2 [1, 2] or panels of candidate genes suspected as being involved in Hereditary Breast Ovarian Cancer (HBOC) [3, 4]. These panels include, in addition to the genes BRCA1 and BRCA2, genes that were found to be associated with a breast cancer risk e.g. PALB2 carriers [5, 6]. The PARP inhibitors act by blocking the BER pathway and promote DNA doublestrand breaks In normal cells, these double-strand breaks are repaired by proteins involved in homologous recombination (HR) which includes the BRCA1 and BRCA2 proteins. The identification of mutations for personalized medicine indications requires a rapid testing for BRCA1 and BRCA2 mutations (4 to 8 weeks)
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
