Marsavela Gabriela, Reid Anna, Gray Elin S, Calapre Leslie
School of Medical and Health Sciences, Edith Cowan University, Joondalup, WA, Australia.
Methods Mol Biol. 2021;2265:247-263. doi: 10.1007/978-1-0716-1205-7_19.
In recent years, circulating tumor DNA (ctDNA) has emerged as a promising prognostic and monitoring biomarker of various cancers, including melanoma. However, sensitive methods are required for its preservation, isolation, and detection. Here we describe a sensitive method for plasma ctDNA isolation using a column-based extraction kit, followed by quantification using a single mutational target with a droplet digital PCR system. This sensitive protocol has been successfully used to quantify diverse mutations present in plasma-derived ctDNA from cancer patients. The full procedure, from blood processing to the analysis of results, takes approximately a day of work.
近年来，循环肿瘤DNA（ctDNA）已成为包括黑色素瘤在内的各种癌症的一种有前景的预后和监测生物标志物。然而，需要敏感的方法来保存、分离和检测它。在这里，我们描述了一种使用基于柱的提取试剂盒进行血浆ctDNA分离的敏感方法，随后使用液滴数字PCR系统对单个突变靶点进行定量。这种敏感的方案已成功用于定量癌症患者血浆来源的ctDNA中存在的各种突变。从血液处理到结果分析的整个过程大约需要一天的工作时间。
