School of Information and Electrical Engineering, Zhejiang University City College, Hangzhou, Zhejiang Province, 310015, China.
Research Centre for Analytical Instrumentation, Institute of Cyber-Systems and Control, State Key Laboratory of Industrial Control Technology, Zhejiang University, Hangzhou, Zhejiang Province, 310027, China.
Talanta. 2022 Dec 1;250:123725. doi: 10.1016/j.talanta.2022.123725. Epub 2022 Jul 8.
Digital PCR is a sensitive detection method, which has important applicability in liquid biopsy through the measurement of ctDNA. However, the current sample pre-processing of ctDNA and the multiplex detection capability of digital PCR have limitations. In view of the above two aspects, we developed a digital PCR chip with multiplex capability and established a direct amplification detection method without nucleic acid extraction. Through the design and processing of the chip, we established a self-priming multiplex digital PCR chip, which can detect 4 targets using single fluorescence. This method can be applied to most digital PCR chips. In addition, we used the plasma of lung cancer patients to establish a direct digital PCR detection method based on the chip, thereby avoiding disadvantages caused by the ctDNA extraction process. As a proof of concept, we prepared blood plasma samples with different concentration of ctDNA to prove the chip's multiplex detection capabilities and the results suggested that this multiplex digital PCR is accurate. Overall, our platform provides a novel and promising option for the detection of ctDNA.
数字 PCR 是一种灵敏的检测方法，通过测量 ctDNA，在液体活检中有重要的应用。然而，目前 ctDNA 的样本预处理和数字 PCR 的多重检测能力存在局限性。针对上述两个方面，我们开发了具有多重检测能力的数字 PCR 芯片，并建立了无需核酸提取的直接扩增检测方法。通过芯片的设计和加工，我们建立了一种自启动多重数字 PCR 芯片，可使用单荧光检测 4 个靶标。该方法可应用于大多数数字 PCR 芯片。此外，我们使用肺癌患者的血浆，基于该芯片建立了一种直接数字 PCR 检测方法，从而避免了 ctDNA 提取过程带来的弊端。作为概念验证，我们用不同浓度的 ctDNA 制备了血浆样本，以证明该芯片的多重检测能力，结果表明该多重数字 PCR 是准确的。总的来说，我们的平台为 ctDNA 的检测提供了一种新颖而有前途的选择。
