Objective: To compare the efficacy and clinical value of high-throughput sequencing (HTS) and Sanger sequencing in detecting ABL kinase domain mutations in patients with chronic myeloid leukemia (CML).
Methods: A total of 198 samples of 147 CML patients from July 2017 to March 2021 in Henan Cancer Hospital were collected and underwent high-throughput sequencing and Sanger sequencing to detect the mutations in ABL kinase domain, and the relevant clinical data were collected for comparative analysis.
Results: The proportion of total mutations and ≥2 mutations detected by high-throughput sequencing were significantly higher than those detected by Sanger sequencing (P =0.01; P =0.046). ≥2 mutations were detected in 22 cases, of which 5 cases (22.7%) had compound mutations. High-throughput sequencing can detect low level mutations that cannot be detected by Sanger sequencing. In 198 samples, 25 (12.6%) were low level mutations, 33 (16.7%) were high level mutations and 10 (5.1%) were mixed high and low level mutations. In the analysis of related clinical factors, the total mutation rate and the low level mutation rate in the optimal period, failure period and warning period were gradually increased (total mutation rate, P =0.016; low level mutation rate, P =0.005). The mutation rate of the samples with additional chromosomal abnormalities was also significantly increased (P =0.009). The mutation rate of patients who received first- and second-line treatment was significantly lower than that of patients who received third- or higher-line treatment (P =0.006). Analysis based on variant allele frequency (VAF) of the mutation site was helpful to visually evaluate the clonal evolution status of TKI-resistance CML cells.
Conclusion: High-throughput sequencing is more sensitive and accurate than Sanger sequencing in mutation detection, which is helpful to accurately and visually evaluate TKI treatment response and optimize treatment strategy for CML.
题目: 基于高通量测序技术检测慢性髓性白血病患者ABL激酶区突变的临床价值.
目的: 对比分析高通量测序法与Sanger测序法检测慢性髓性白血病（CML）患者ABL激酶区突变的效能及临床价值。.
方法: 采集2017年7月至2021年3月于河南省肿瘤医院就诊的147例CML患者的198例次样本，同时进行高通量测序和Sanger测序，检测ABL激酶区突变，并收集相关临床数据进行对比分析。.
结果: 高通量测序法检出总体突变比例和≥2个突变的比例明显高于Sanger测序法（P =0.01；P =0.046）。22例次检出≥2个突变，其中发现5例次（22.7%）复合突变。高通量测序法可检出Sanger法无法检出的低比例突变。198例次样本中检出低比例突变25例次（12.6%），高比例突变33例次（16.7%）和高低比例混合突变10例次（5.1%）。相关临床因素分析中，最佳反应期、失败期和警告期突变检出比例和发生低比例突变占比均逐渐升高（突变比例，P =0.016；低比例突变，P =0.005），伴随附加染色体异常出现突变的比例也明显升高（P =0.009）。接受一线、二线治疗的患者检出突变的比例明显低于接受三线及以上治疗的患者（P =0.006）。基于突变位点的VAF值分析有助于可视化评估耐药CML细胞的克隆演变状态。.
结论: 高通量测序法较Sanger测序法检测突变更加敏感、准确，有助于临床精准、可视化评估TKI治疗反应和优化CML治疗策略。.
Keywords: ABL kinase domain mutations; Sanger sequencing; chronic myeloid leukemia; high-throughput sequencing; tyrosine kinase inhibitors.