Therapy Monitoring of EGFR-Positive Non-Small-Cell Lung Cancer Patients Using ddPCR Multiplex Assays
Therapy Monitoring of EGFR-Positive Non-Small-Cell Lung Cancer Patients Using ddPCR Multiplex Assays
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DOI：
10.1016/j.jmoldx.2021.01.003
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发表时间：
2021-03-22
影响因子：
4.1
通讯作者：
Deiman, Birgit
中科院分区：
文献类型：
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作者：
de Kock, Remco;van den Borne, Ben;Deiman, Birgit
晚期非小细胞肺癌患者EGFR致敏和耐药突变的检测对于EGFR酪氨酸激酶抑制剂治疗的选择和监测具有重要意义。液滴数字PCR （ddPCR）多重检测允许灵敏和同时检测无细胞DNA （cfDNA）中的多个突变，所需的提取物最少，成本较低。使用一种新型的ddPCR五联体试验筛选EGFR酪氨酸激酶抑制剂致敏突变Ex19Del、L858R、L861Q、G719S和S768I。在治疗期间，对检测呈阳性的患者进行egfr致敏突变和两种egfr耐药突变（T790M和C797S）的监测，使用ddPCR监测三重试验。ddPCR多重检测能够可靠地检测每个突变，其分数丰度至少为0.1%。对6例患者进行纵向数据分析，ddPCR结果能很好地反映病程和放射学反应。本研究证实了cfDNA上的ddPCR支持诊断和治疗选择，并表明cfDNA上的ddPCR多重检测可以成为非小细胞肺癌患者治疗监测的一种有价值的附加诊断工具。
The detection of EGFR-sensitizing and EGFR-resistance mutations in advanced non-small-cell lung cancer patients is important for the selection and monitoring of EGFR tyrosine-kinase inhibitor therapy. Droplet digital PCR (ddPCR) multiplex assays allow for sensitive and simultaneous detection of multiple mutations in cell-free DNA (cfDNA) with a minimum of extract needed and at lower cost. Patients were screened for the EGFR tyrosine-kinase inhibitor-sensitizing mutations Ex19Del, L858R, L861Q, G719S, and S768I using a novel ddPCR pentaplex assay. Patients who tested positive subsequently were monitored during treatment for the EGFR-sensitizing mutation and two EGFR-resistance mutations, T790M and C797S, using a ddPCR monitor triplex assay. The ddPCR multiplex assays enabled reliable detection of each mutation with a fractional abundance of at least 0.1%. For six patients, longitudinal data were analyzed and the ddPCR results provided a good reflection of the course of the disease and radiologic response. This study confirms that ddPCR on cfDNA supports the diagnosis and therapy selection, and shows that ddPCR multiplex assays on cfDNA could be a valuable additional diagnostic tool for therapy monitoring of non-small-cell lung cancer patients.