Kluska Anna, Balabas Aneta, Paziewska Agnieszka, Kulecka Maria, Nowakowska Dorota, Mikula Michal, Ostrowski Jerzy
Department of Genetics, Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, 02-781, Warsaw, Poland.
Department of Gastroenterology, Hepatology and Clinical Oncology, Medical Center for Postgraduate Education, 01-813, Warsaw, Poland.
BMC Med Genomics. 2015 May 7;8:19. doi: 10.1186/s12920-015-0092-2.
Targeted PCR-based genetic testing for BRCA1 and BRCA2 can be performed at a lower cost than full gene testing; however, it may overlook mutations responsible for familial breast and/or ovarian cancers. In the present study, we report the utility of next generation sequencing (NGS) to identify new pathogenic variants of BRCA1/2.
BRCA1 and BRCA2 exons were amplified using the Ion AmpliSeq BRCA1/2 Panel and sequenced on the Ion Torrent PGM sequencer in 512 women with familial and/or only early onset breast and/or ovarian cancers who were negative for selected BRCA1/2 mutations.
146 single nucleotide variants (SNVs) and 32 indels were identified. Of them, 14 SNVs and 17 indels were considered as pathogenic or likely pathogenic. One and 18 pathogenic mutations had been detected previously in the Polish and other populations, respectively, and 12 deleterious mutations were previously unknown. Eight mutations were recurrent; Q563X (BRCA1), N3124I (BRCA2) and c.4516delG (BRCA1) were found in eight, six and four patients, respectively, and two other mutations (c.9118-2A > G and c.7249delCA in BRCA2) were detected in three patients each. Altogether, BRCA1/2 pathogenic mutations were identified in 52 out of 512 (10%) patients.
NGS substantially improved the detection rates of a wide spectrum of mutations in Polish patients with familial breast and/or ovarian cancer. Although targeted screening for specific BRCA1 mutations can be offered to all Polish breast or ovarian cancer patients, NGS-based testing is justified in patients with breast or ovarian cancer likely related to BRCA1/2 who test negative for the selected BRCA1/2 pathogenic mutations.
基于靶向聚合酶链反应（PCR）的BRCA1和BRCA2基因检测成本低于全基因检测；然而，它可能会遗漏导致家族性乳腺癌和/或卵巢癌的突变。在本研究中，我们报告了下一代测序（NGS）在鉴定BRCA1/2新的致病变异方面的效用。
使用Ion AmpliSeq BRCA1/2 Panel扩增BRCA1和BRCA2外显子，并在Ion Torrent PGM测序仪上对512名患有家族性和/或仅早发性乳腺癌和/或卵巢癌且选定的BRCA1/2突变检测为阴性的女性进行测序。
共鉴定出146个单核苷酸变异（SNV）和32个插入缺失。其中，14个SNV和17个插入缺失被认为是致病的或可能致病的。分别在波兰人群和其他人群中先前检测到1个和18个致病突变，还有12个有害突变此前未知。8个突变是复发性的；Q563X（BRCA1）、N3124I（BRCA2）和c.4516delG（BRCA1）分别在8例、6例和4例患者中发现，另外两个突变（BRCA2中的c.9118-2A＞G和c.7249delCA）在3例患者中各有发现。总共在512例患者中的52例（10%）中鉴定出BRCA1/2致病突变。
NGS显著提高了波兰家族性乳腺癌和/或卵巢癌患者中广泛突变的检测率。虽然可以为所有波兰乳腺癌或卵巢癌患者提供针对特定BRCA1突变的靶向筛查，但对于可能与BRCA1/2相关且选定的BRCA1/2致病突变检测为阴性的乳腺癌或卵巢癌患者，基于NGS的检测是合理的。
