Rajkumar Thangarajan, Meenakumari Balaiah, Mani Samson, Sridevi Veluswami, Sundersingh Shirley
Department of Molecular Oncology, Cancer Institute (WIA), Chennai, India E-mail :
Asian Pac J Cancer Prev. 2015;16(13):5211-7. doi: 10.7314/apjcp.2015.16.13.5211.
We earlier used PCR-dHPLC for mutation analysis of BRCA1 and BRCA2. In this article we report application of targeted resequencing of 30 genes involved in hereditary cancers.
A total of 91 patient samples were analysed using a panel of 30 genes in the Illumina HiScan SQ system. CLCBio was used for mapping reads to the reference sequences as well as for quality-based variant detection. All the deleterious mutations were then reconfirmed using Sanger sequencing. Kaplan Meier analysis was conducted to assess the effect of deleterious mutations on disease free and overall survival.
Seventy four of the 91 samples had been run earlier using the PCR-dHPLC and no deleterious mutations had been detected while 17 samples were tested for the first time. A total of 24 deleterious mutations were detected, 11 in BRCA1, 4 in BRCA2, 5 in p53, one each in RAD50, RAD52, ATM and TP53BP1. Some 19 deleterious mutations were seen in patients who had been tested earlier with PCR-dHPLC [19/74] and 5/17 in the samples tested for the first time, Together with our earlier detected 21 deleterious mutations in BRCA1 and BRCA2, we now had 45 mutations in 44 patients. BRCA1c.68_69delAG;p.Glu23ValfsX16 mutation was the most common, seen in 10/44 patients. Kaplan Meier survival analysis did not show any difference in disease free and overall survival in the patients with and without deleterious mutations.
The NGS platform is more sensitive and cost effective in detecting mutations in genes involved in hereditary breast and/or ovarian cancers.
我们之前使用聚合酶链反应-变性高效液相色谱法（PCR-dHPLC）对BRCA1和BRCA2进行突变分析。在本文中，我们报告了对30个遗传性癌症相关基因进行靶向重测序的应用情况。
使用Illumina HiScan SQ系统中的一组30个基因对总共91份患者样本进行分析。使用CLCBio将读取序列映射到参考序列，并基于质量进行变异检测。然后使用桑格测序法对所有有害突变进行再次确认。进行卡普兰-迈耶分析以评估有害突变对无病生存期和总生存期的影响。
91份样本中的74份之前已使用PCR-dHPLC进行检测，未检测到有害突变，而17份样本是首次检测。总共检测到24个有害突变，其中11个在BRCA1中，4个在BRCA2中，5个在p53中，RAD50、RAD52、ATM和TP53BP1各有1个。在之前使用PCR-dHPLC检测的患者中发现了约19个有害突变[19/74]，在首次检测的样本中有5/17个。连同我们之前在BRCA1和BRCA2中检测到的21个有害突变，我们现在在44名患者中发现了45个突变。BRCA1c.68_69delAG;p.Glu23ValfsX16突变最为常见，在10/44名患者中出现。卡普兰-迈耶生存分析未显示有无有害突变的患者在无病生存期和总生存期方面存在任何差异。
新一代测序（NGS）平台在检测遗传性乳腺癌和/或卵巢癌相关基因的突变方面更敏感且更具成本效益。
