Molecular Barcode Sequencing for Highly Sensitive Detection of Circulating Tumor DNA in Patients with Esophageal Squamous Cell Carcinoma
Molecular Barcode Sequencing for Highly Sensitive Detection of Circulating Tumor DNA in Patients with Esophageal Squamous Cell Carcinoma
复制标题
DOI：
10.1159/000504808
复制
发表时间：
2020-04-01
期刊：
影响因子：
3.5
通讯作者：
Doki, Yuichiro
中科院分区：
文献类型：
--
作者：
Hagi, Takaomi;Kurokawa, Yukinori;Doki, Yuichiro
介绍：使用分子条形码（MB）的下一代测序（NGS）是一种新方法，可以在相对广泛的基因中高度灵敏地检测循环肿瘤DNA（ctDNA）。目的：探讨NGS联合MB检测食管鳞状细胞癌（esophageal squamous cell carcinoma，ESCC）患者ctDNA的价值。方法：对5例食管鳞癌患者进行了术前治疗，随后进行了食管切除术。使用不含MB的NGS分析治疗过程中每个时间点从肿瘤组织和血浆提取的DNA中TP 53突变的频率。在1例患者中，使用NGS和MB进行了额外分析，以比较灵敏度并评价这种新方法的临床实用性。结果：5例食管鳞癌中3例肿瘤组织中存在TP53突变。1例患者术前血浆中突变等位基因频率为1.97%，术前治疗后降至0.09%。NGS检测ctDNA的最大背景误差率为3.22%，而NGS检测ctDNA的最大背景误差率为0.08%，说明NGS检测ctDNA的灵敏度明显高于NGS。在1例术后半年复发的患者中，仅NGS与MB在术后4周仍能检测到ctDNA，频率为0.20%。结论：带有MB的NGS能够全面且高度灵敏地检测ESCC患者的ctDNA。这种新的方法可能有助于ESCC的临床诊断。
Introduction:Next-generation sequencing (NGS) with molecular barcodes (MB) is a novel method that enables the highly sensitive detection of circulating tumor DNA (ctDNA) in a relatively wide range of genes. Objective:The aim of this study was to examine the utility of NGS with MB for detecting ctDNA in patients with esophageal squamous cell carcinoma (ESCC). Methods:Five patients with ESCC who underwent preoperative treatment followed by esophagectomy were examined. The frequency of TP53 mutations in DNA extracted from tumor tissue and plasma at each time point during the treatment course was analyzed using NGS without MB. In 1 patient, additional analysis using NGS with MB was conducted to compare the sensitivities and to evaluate the clinical utility of this novel method. Results:TP53 mutations in tumor tissue were identified in 3 of 5 patients with ESCC. In 1 patient, the mutational allele frequency in plasma was 1.97% before preoperative treatment, and decreased to 0.09% after preoperative treatment. As the maximum frequency of background errors were 3.22% using NGS without MB and 0.08% with MB, which indicated that the sensitivity of ctDNA detection using NGS with MB was much higher than without MB. In 1 patient who had recurrence half a year after surgery, only NGS with MB could detect ctDNA even at 4 weeks after surgery, at a frequency of 0.20%. Conclusions:NGS with MB enabled comprehensive and highly sensitive detection of ctDNA in a patient with ESCC. This novel method may be useful for the clinical diagnosis of ESCC.