Department of Biomedical Engineering, School of Life Sciences, Ulsan National Institute of Science and Technology (UNIST), Ulsan 44919, Republic of Korea.
Lab Chip. 2018 May 1;18(9):1320-1329. doi: 10.1039/c8lc00165k.
The potential utility of circulating tumour DNA (ctDNA) in patient blood for cancer diagnostics and real-time monitoring of disease progression is highly recognized. However, the lack of automated and efficient methods for cell-free DNA (cfDNA) isolation from peripheral blood has remained a challenge for broader acceptance of liquid biopsy in general clinical settings. Here, we demonstrate a lab-on-a-disc system equipped with newly developed, electromagnetically actuated, and reversible diaphragm valves that allows fully automated and rapid (<30 min) isolation of cfDNA from whole blood (>3 ml) to achieve high detection sensitivity by minimizing the degradation of fragile ctDNA as well as contamination of wild-type DNA from abundant blood cells. As a proof of concept study, we used the lab-on-a-disc to isolate cfDNA from patients with non-small cell lung cancer and successfully detected epidermal growth factor receptor gene mutations (L858R, T790M) during targeted drug therapy. The proposed lab-on-a-disc enables a fully automated, rapid, and point-of-care cfDNA enrichment starting from whole blood to facilitate the wide use of liquid biopsy in routine clinical practice.
循环肿瘤 DNA(ctDNA)在患者血液中用于癌症诊断和实时监测疾病进展的潜在效用已得到广泛认可。然而,缺乏从外周血中自动且高效地分离游离 DNA(cfDNA)的方法,这一直是液体活检在一般临床环境中更广泛应用的一个挑战。在这里,我们展示了一种配备了新型电磁驱动、可逆隔膜阀的碟式芯片实验室系统,该系统允许从全血(>3 毫升)中全自动且快速(<30 分钟)分离 cfDNA,从而通过最小化脆弱的 ctDNA 降解以及从丰富的血细胞中污染野生型 DNA 来实现高检测灵敏度。作为概念验证研究,我们使用碟式芯片实验室从非小细胞肺癌患者中分离 cfDNA,并在靶向药物治疗期间成功检测到表皮生长因子受体基因突变(L858R、T790M)。该碟式芯片实验室能够从全血开始全自动、快速、即时地富集 cfDNA,从而促进液体活检在常规临床实践中的广泛应用。