Department of Molecular Oncology, Quest Diagnostics Nichols Institute, Chantilly, Virginia.
J Mol Diagn. 2020 Aug;22(8):994-1007. doi: 10.1016/j.jmoldx.2020.04.213. Epub 2020 May 29.
Next-generation sequencing (NGS) technologies have come of age as preferred technologies for screening of genomic variants of pathologic and therapeutic potential. Because of their capability for high-throughput and massively parallel sequencing, they can screen for a variety of genomic changes in multiple samples simultaneously. This has made them platforms of choice for clinical testing of solid tumors and hematological malignancies. Consequently, they are increasingly replacing conventional technologies, such as Sanger sequencing and pyrosequencing, expression arrays, real-time PCR, and fluorescence in situ hybridization methods, for routine molecular testing of tumors. However, one limitation of routinely used NGS technologies is the inability to detect low-level genomic variants with high accuracy. This can be attributed to the frequent occurrence of low-level sequencing errors and artifacts in NGS workflow that need specialized approaches to be identified and eliminated. This review focuses on the origins and nature of these artifacts and recent improvements in the NGS technologies to overcome them to facilitate accurate high-sensitive detection of low-level mutations. Potential applications of high-sensitive NGS in oncology and comparisons with non-NGS technologies of similar capabilities are also summarized.
下一代测序（NGS）技术已经成为具有病理和治疗潜力的基因组变异筛选的首选技术。由于其高通量和大规模平行测序的能力，它们可以同时对多种样本中的多种基因组变化进行筛选。这使得它们成为实体瘤和血液恶性肿瘤临床检测的首选平台。因此，它们正在逐渐取代传统技术，如 Sanger 测序和焦磷酸测序、表达谱芯片、实时 PCR 和荧光原位杂交方法，用于肿瘤的常规分子检测。然而，常规使用的 NGS 技术的一个局限性是无法高精度地检测低水平的基因组变异。这可以归因于 NGS 工作流程中低频测序错误和伪影的频繁发生，需要专门的方法来识别和消除这些错误和伪影。本综述重点介绍了这些伪影的来源和性质，以及最近在克服这些伪影以促进低水平突变的准确高灵敏度检测方面的 NGS 技术的改进。还总结了高灵敏度 NGS 在肿瘤学中的潜在应用以及与类似功能的非 NGS 技术的比较。
