Badoer Cindy, Garrec Céline, Goossens Dirk, Ellison Gillian, Mills John, Dzial Mélina, El Housni Hakim, Berwouts Sarah, Concolino Paola, Guibert-Le Guevellou Virginie, Delnatte Capucine, Del Favero Jurgen, Capoluongo Ettore, Bézieau Stéphane
Laboratoire de Génétique Moléculaire, Clinique Universitaire de Bruxelles-Hôpital Erasme-Université Libre de Bruxelles (CUB-Erasme-ULB), Brussels, Belgium.
Institut de Biologie, Laboratoire de Génétique Moléculaire, Service de Génétique Médicale, CHU Nantes, Nantes, France.
Oncotarget. 2016 Dec 6;7(49):81357-81366. doi: 10.18632/oncotarget.12877.
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.
新一代测序（NGS）为检测与遗传性乳腺癌和卵巢癌（HBOC）相关的BRCA1和BRCA2基因中的突变提供了新方法。在肿瘤发生和手术（双侧乳房切除术、卵巢切除术）咨询方面，寻找BRCA1和BRCA2基因中的种系突变具有重要意义。随着聚ADP核糖聚合酶（PARP）抑制剂奥拉帕尼现已可用于治疗特定形式的卵巢癌和BRCA突变患者，检测肿瘤材料中的BRCA突变对于治疗决策变得越来越重要。分子遗传学实验室应开发可靠且灵敏的技术，以对BRCA1和BRCA2基因进行全面分析。由于BRCA1/2基因编码序列的长度、缺乏热点突变，尤其是从福尔马林固定石蜡包埋（FFPE）组织中获得的DNA质量较低，这是一项挑战。因此，许多分析无法解读，并且并不总是能为临床医生提供结果，从而限制了患者的最佳治疗管理。对于一些实验室来说，新鲜冷冻组织（FFT）的可用性以及从中提取的DNA的优异质量提供了一种替代方案。出于这个原因，我们使用Multiplicom公司的BRCA MASTR Dx检测方法，结合用于Illumina MiSeq的MID，对一组97个来自FFT的DNA样本进行了BRCA1和BRCA2突变检测评估。我们获得了所有测试样本的可解读NGS结果，并显示出>99.7%的灵敏度、特异性和准确性。
