左冉, 苏雨栋, 孟昭婷, 王心悦, 林丽, 张翠翠, 陈金良, 王雅杰, 刘萍萍, 于津浦, 李凯, 陈鹏. SM-PCR技术检测血浆ctDNA在晚期肺腺癌患者治疗中的应用[J]. 中国肿瘤临床, 2019, 46(8): 384-388. DOI: 10.3969/j.issn.1000-8179.2019.08.189
引用本文: 左冉, 苏雨栋, 孟昭婷, 王心悦, 林丽, 张翠翠, 陈金良, 王雅杰, 刘萍萍, 于津浦, 李凯, 陈鹏. SM-PCR技术检测血浆ctDNA在晚期肺腺癌患者治疗中的应用[J]. 中国肿瘤临床, 2019, 46(8): 384-388. DOI: 10.3969/j.issn.1000-8179.2019.08.189
shu
Zuo Ran, Su Yudong, Meng Zhaoting, Wang Xinyue, Lin Li, Zhan Cuicui, Chen Jinliang, Wang Yajie, Liu Pingping, Yu Jinpu, Li Kai, Chen Peng. Application of SM-PCR to detect plasma ctDNA in the treatment of patients with advanced lung adenocarcinoma[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2019, 46(8): 384-388. DOI: 10.3969/j.issn.1000-8179.2019.08.189
Citation: Zuo Ran, Su Yudong, Meng Zhaoting, Wang Xinyue, Lin Li, Zhan Cuicui, Chen Jinliang, Wang Yajie, Liu Pingping, Yu Jinpu, Li Kai, Chen Peng. Application of SM-PCR to detect plasma ctDNA in the treatment of patients with advanced lung adenocarcinoma[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2019, 46(8): 384-388. DOI: 10.3969/j.issn.1000-8179.2019.08.189
shu

SM-PCR技术检测血浆ctDNA在晚期肺腺癌患者治疗中的应用

Application of SM-PCR to detect plasma ctDNA in the treatment of patients with advanced lung adenocarcinoma

    摘要
  • 摘要:
      目的  探讨单分子PCR（single molecule-PCR，SM-PCR）技术检测血浆ctDNA在晚期肺腺癌患者治疗中的应用。
      方法  分析2017年6月至2018年5月就诊于天津医科大学肿瘤医院的晚期肺腺癌患者30例，使用SM-PCR技术富集血液样本目标基因（EGFR、KRAS、BRAF、ALK、HER2、TP53）区域的ctDNA片段，构建测序文库，进行高通量测序；使用基于扩增阻滞突变系统（amplication refractory mutation system，ARMS）实时荧光PCR法进行肿瘤组织样本EGFR检测，比较血浆与组织表皮生长因子受体（epidermal growth factor receptor，EGFR）突变检测结果的一致性。
      结果  SM-PCR与ARMS-PCR两种方法检测结果一致性较好（Kappa= 0.867，P < 0.001）；McNemar检验亦提示检测结果差异无统计学意义（P=0.500）。
      结论  SM-PCR技术可用于血浆EGFR突变检测，其目标检测位点更全面，且可同时检测多种突变，分析结果更为精细，可实现绝对定量。

     

    Abstract:
      Objective  To investigate the application of single-molecule PCR (SM-PCR) in the detection of plasma ctDNA for the treatment of patients with advanced lung adenocarcinoma.
      Methods  In total, 30 patients diagnosed with advanced lung adenocarcinoma were enrolled between June 2017 and May 2018. ctDNA fragments of the target genes (EGFR, KRAS, BRAF, ALK, HER2, and TP53) from the blood samples were enriched by SM-PCR, and DNA libraries were prepared. Finally, a high-throughput sequencing was performed. The EGFR detection of tumor tissue samples was performed using real-time fluorescence PCR based on the amplification refractory mutation system (ARMS) and consistency in the results of EGFR mutation detection in the plasma and tissue was compared.
      Results  The results of both the methods were consistent (Kappa=0.867, P < 0.001). The McNemar's test also indicated that the results are not statistically different (P=0.500).
      Conclusions  SM-PCR can be used for the detection of plasma EGFR mutations. The target detection sites are more comprehensive and multiple mutations can be detected at the same time. Results of the analysis are more precise and can be absolutely quantified.

     

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