title: Performance of multiplicom's BRCA MASTR Dx kit on the detection of BRCA1 and BRCA2 mutations in fresh frozen ovarian and breast tumor samples.
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.
authors: Laboratoire de Génétique Moléculaire, Clinique Universitaire de Bruxelles-Hôpital Erasme-Université Libre de Bruxelles (CUB-Erasme-ULB), Brussels, Belgium.
authors: Cindy
authors: C
authors: Badoer
authors: Institut de Biologie, Laboratoire de Génétique Moléculaire, Service de Génétique Médicale, CHU Nantes, Nantes, France.
authors: Céline
authors: C
authors: Garrec
authors: Multiplicom N.V., Niel, Belgium.
authors: Dirk
authors: D
authors: Goossens
authors: AstraZeneca, Personalised Healthcare and Biomarkers, Alderley Park, Macclesfield, UK.
authors: Gillian
authors: G
authors: Ellison
authors: AstraZeneca, Personalised Healthcare and Biomarkers, Alderley Park, Macclesfield, UK.
authors: John
authors: J
authors: Mills
authors: Laboratoire de Génétique Moléculaire, Clinique Universitaire de Bruxelles-Hôpital Erasme-Université Libre de Bruxelles (CUB-Erasme-ULB), Brussels, Belgium.
authors: Mélina
authors: M
authors: Dzial
authors: Laboratoire de Génétique Moléculaire, Clinique Universitaire de Bruxelles-Hôpital Erasme-Université Libre de Bruxelles (CUB-Erasme-ULB), Brussels, Belgium.
authors: Hakim
authors: H
authors: El Housni
authors: Multiplicom N.V., Niel, Belgium.
authors: Sarah
authors: S
authors: Berwouts
authors: Laboratory of Clinical Molecular and Personalized Diagnostics, Foundation Policlinico Gemelli and Catholic University of Rome, Italy.
authors: Paola
authors: P
authors: Concolino
authors: Institut de Biologie, Laboratoire de Génétique Moléculaire, Service de Génétique Médicale, CHU Nantes, Nantes, France.
authors: Virginie
authors: V
authors: Guibert-Le Guevellou
authors: Institut de Biologie, Laboratoire de Génétique Moléculaire, Service de Génétique Médicale, CHU Nantes, Nantes, France.
authors: Capucine
authors: C
authors: Delnatte
authors: Multiplicom N.V., Niel, Belgium.
authors: Jurgen
authors: J
authors: Del Favero
authors: Laboratory of Clinical Molecular and Personalized Diagnostics, Foundation Policlinico Gemelli and Catholic University of Rome, Italy.|Molipharma and Giovanni Paolo II Foundation, Campobasso, Italy.
authors: Ettore
authors: E
authors: Capoluongo
authors: Institut de Biologie, Laboratoire de Génétique Moléculaire, Service de Génétique Médicale, CHU Nantes, Nantes, France.
authors: Stéphane
authors: S
authors: Bézieau
mesh_terms: D019313:BRCA1 Protein / Q000235:genetics*; D024682:BRCA2 Protein / Q000235:genetics*; D014408:Biomarkers, Tumor / Q000235:genetics*; D001943:Breast Neoplasms / Q000235:genetics* / Q000473:pathology; D004252:DNA Mutational Analysis / Q000379:methods*; D005060:Europe; D005260:Female; D005615:Freezing; D005787:Gene Frequency; D020022:Genetic Predisposition to Disease; D040941:Heredity; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction; D009154:Mutation; D010051:Ovarian Neoplasms / Q000235:genetics* / Q000473:pathology; D011237:Predictive Value of Tests; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results; D013048:Specimen Handling / Q000379:methods*
chemical_list: D019313:BRCA1 Protein; C492913:BRCA1 protein, human; D024682:BRCA2 Protein; C551750:BRCA2 protein, human; D014408:Biomarkers, Tumor; D011933:Reagent Kits, Diagnostic
keywords: BRCA1-BRCA2; fresh frozen tumors; next generation sequencing; olaparib; ovarian carcinoma