School of Dentistry, University of California, Los Angeles, CA 90095, USA.
Department of Physics, University of California, Los Angeles, CA 90095, USA.
Int J Mol Sci. 2023 Jun 20;24(12):10387. doi: 10.3390/ijms241210387.
Liquid biopsy is a rapidly emerging field that involves the minimal/non-invasive assessment of signature somatic mutations through the analysis of circulating tumor DNA (ctDNA) shed by tumor cells in bodily fluids. Broadly speaking, the unmet need in liquid biopsy lung cancer detection is the lack of a multiplex platform that can detect a mutation panel of lung cancer genes using a minimum amount of sample, especially for ultra-short ctDNA (usctDNA). Here, we developed a non-PCR and non-NGS-based single-droplet-based multiplexing microsensor technology, "Electric-Field-Induced Released and Measurement (EFIRM) Liquid Biopsy" (m-eLB), for lung cancer-associated usctDNA. The m-eLB provides a multiplexable assessment of usctDNA within a single droplet of biofluid in only one well of micro-electrodes, as each electrode is coated with different probes for the ctDNA. This m-eLB prototype demonstrates accuracy for three tyrosine-kinase-inhibitor-related EGFR target sequences in synthetic nucleotides. The accuracy of the multiplexing assay has an area under the curve (AUC) of 0.98 for L858R, 0.94 for Ex19 deletion, and 0.93 for T790M. In combination, the 3 EGFR assay has an AUC of 0.97 for the multiplexing assay.
液体活检是一个快速发展的领域，通过分析肿瘤细胞在体液中释放的循环肿瘤 DNA（ctDNA），对特征性体细胞突变进行最小程度的非侵入性评估。广义上讲，液体活检肺癌检测中未满足的需求是缺乏一种能够使用最少量样本检测肺癌基因突变面板的多重平台，特别是对于超短 ctDNA（usctDNA）而言。在这里，我们开发了一种非 PCR 和非 NGS 基于单液滴的多重微传感器技术，“电场诱导释放和测量（EFIRM）液体活检”（m-eLB），用于肺癌相关的 usctDNA。m-eLB 提供了在单个微电极孔中的单个生物流体液滴内对 usctDNA 的多重评估，因为每个电极都涂有用于 ctDNA 的不同探针。该 m-eLB 原型在合成核苷酸中证明了针对三个酪氨酸激酶抑制剂相关的 EGFR 靶序列的准确性。多重分析的准确性对于 L858R 的 AUC 为 0.98，对于 Ex19 缺失的 AUC 为 0.94，对于 T790M 的 AUC 为 0.93。三者结合，3 个 EGFR 分析对于多重分析的 AUC 为 0.97。
