A Rapid and Sensitive Next-Generation Sequencing Method to Detect RB1 Mutations Improves Care for Retinoblastoma Patients and Their Families
A Rapid and Sensitive Next-Generation Sequencing Method to Detect RB1 Mutations Improves Care for Retinoblastoma Patients and Their Families
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DOI：
10.1016/j.jmoldx.2016.02.006
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发表时间：
2016-07-01
影响因子：
4.1
通讯作者：
Triche, Timothy J.
中科院分区：
文献类型：
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作者：
Li, Wenhui L.;Buckley, Jonathan;Triche, Timothy J.
视网膜母细胞瘤是一种儿童眼部恶性肿瘤，可导致视力，眼睛，有时甚至生命的丧失。肿瘤是由肿瘤抑制基因RB1的两个等位基因的失活突变引起的，或者很少由MYCN扩增引起的。及时识别血液样本中的种系RB 1突变或肿瘤中的体细胞RB 1突变或MYCN扩增对于视网膜母细胞瘤患者及其家人的有效护理和管理非常重要。然而，目前彻底测试RB1突变的程序复杂且冗长。在本文中，我们报告了一种新一代基于测序的方法，该方法能够在单个平台上检测整个RB1基因的点突变、小插入缺失和大缺失或重复以及MYCN基因的扩增。从DNA提取到临床解释仅需3天，可实现视网膜母细胞瘤的早期分子诊断和最佳治疗结果。该方法还可以检测血液样品中常规桑格测序可能遗漏的低水平镶嵌突变。此外，它可以区分RB1突变和MYCN扩增驱动的视网膜母细胞瘤。这种快速、全面、灵敏的检测RB1突变和MYCN扩增的方法可以很容易地识别RB1突变携带者，从而改善视网膜母细胞瘤患者及其家属的管理和遗传咨询。
Retinoblastoma is a childhood eye malignancy that can lead to the loss of vision, eye(s), and sometimes life. The tumors are initiated by inactivating mutations in both alleles of the tumor-suppressor gene, RB1, or, rarely, by MYCN amplification. Timely identification of a germline RB1 mutation in blood samples or either somatic RB1 mutation or MYCN amplification in tumors is important for effective care and management of retinoblastoma patients and their families. However, current procedures to thoroughly test RB1 mutations are complicated and Lengthy. Herein, we report a next-generation sequencing-based method capable of detecting point mutations, small indels, and Large deletions or duplications across the entire RB1 gene and amplification of MYCN gene on a single platform. From DNA extraction to clinical interpretation requires only 3 days, enabling early molecular diagnosis of retinoblastoma and optimal treatment outcomes. This method can also detect Low-Level mosaic mutations in blood samples that can be missed by routine Sanger sequencing. In addition, it can differentiate between RB1 mutation- and MYCN amplification-driven retinoblastomas. This rapid, comprehensive, and sensitive method for detecting RB1 mutations and MYCN amplification can readily identify RB1 mutation carriers and thus improve the management and genetic counseling for retinoblastoma patients and their families.