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.

新一代测序 (NGS) 技术已经成为筛选具有病理和治疗潜力的基因组变异的首选技术。由于它们具有高通量和大规模并行测序的能力，它们可以同时筛选多个样本中的各种基因组变化。这使它们成为实体瘤和血液系统恶性肿瘤临床测试的首选平台。因此，它们越来越多地取代传统技术，例如 Sanger 测序和焦磷酸测序、表达阵列、实时 PCR 和原位荧光杂交方法，用于肿瘤的常规分子检测。然而，常规使用的 NGS 技术的一个限制是无法高精度检测低水平的基因组变异。这可归因于 NGS 工作流程中经常发生低级测序错误和伪影，需要专门的方法来识别和消除这些错误。本综述重点关注这些伪影的起源和性质，以及 NGS 技术的最新改进，以克服它们以促进对低水平突变的准确高灵敏度检测。还总结了高灵敏度 NGS 在肿瘤学中的潜在应用以及与具有类似功能的非 NGS 技术的比较。