Application of next-generation sequencing in the detection of low-abundance mutations.

Mutation accumulation in somatic cells contributes to cancer development, aging and many non-malignant diseases. The true mutation frequency in normal cells is extremely low, which presents a challenge in detecting these mutations at such low frequencies. The emergence of next-generation sequencing (NGS) technology enables direct detection of rare mutations across the entire genome of any species. This breakthrough overcomes numerous limitations of traditional mutation detection techniques that rely on specific detection models and sites. However, conventional NGS is limited in its application for detecting low-frequency mutations due to its high sequencing error rate. To address this challenge, high-accuracy NGS sequencing techniques based on molecular consensus sequencing strategies have been developed. These techniques have the ability to correct sequencing errors, resulting in error rates lower than 10-7, are expected to serve as effective tools for low-frequency mutation detection. Error-corrected NGS (ecNGS) techniques hold great potential in various areas, including safety evaluation and research on environmental mutagens, risk assessment of cell and gene therapy drugs, population health risk monitoring, and fundamental research in life sciences. This review highlights a comprehensive review of the research progress in low-frequency mutation detection techniques based on NGS, and provides a glimpse into their potential applications. It also offers an outlook on the potential applications of these techniques, thereby providing valuable insights for further development, research, and application of this technology in relevant fields.体细胞突变的累积与衰老、肿瘤及多种疾病的发生密切相关。在正常组织细胞中，基因组中自发突变和诱发突变的变异等位基因频率极低，对这类低频突变的检测一直面临挑战。第二代和第三代高通量测序(next-generation sequencing，NGS)技术的出现，可以实现任意物种全基因组上变异的直接检测，克服传统突变检测技术的诸多局限性。但是常规NGS由于测序错误率较高从而限制了其在低频突变检测上的应用，基于分子一致性测序策略进行错误矫正的高准确性NGS测序技术作为有效的低频突变检测工具，有望在环境诱变剂的评价与研究、细胞与基因治疗药物风险评估、人群健康风险监测和生命科学基础研究领域发挥重要作用。本文对比经典突变检测方法，对基于NGS的低频突变检测技术研究进展进行综述，并对应用前景进行展望，以期为该技术的进一步开发、研究和在相关领域的应用提供参考。.