微流控冲击印刷技术实现的液滴数字 PCR 用于绝对基因定量。
Droplet digital PCR enabled by microfluidic impact printing for absolute gene quantification.
机构信息
Department of Precision Machinery & Precision Instrumentation, University of Science & Technology of China, Hefei, Anhui, 230027, China; Key Laboratory of Precision Scientific Instrumentation of Anhui Higher Education Institutes, University of Science and Technology of China, Hefei, Anhui, 230027, China.
Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, Guangdong, 518055, China.
出版信息
Talanta. 2020 May 1;211:120680. doi: 10.1016/j.talanta.2019.120680. Epub 2019 Dec 28.
Digital PCR enabled high-sensitivity and quantitative measurements of rare biological variants. A new digital droplet-enabled PCR technology was introduced in this paper, which partitioned genetic targets into a planar nanoliter droplet array by using a microfluidic impact printer (MIP) with a disposable microfluidic chip. The accuracy of this MIP-enabled PCR technology was verified by detecting a series of concentration gradients of GAPDH gene across spanning four orders of magnitude (from 0.464 copies/μL to 464 copies/μL). Furthermore, this technology was applied to detect the expressions of p53 gene in colon cancer tissues and adjacent nontumorous tissues, from which the copies of the nucleic acids could be absolute-quantitatively determined. The outcomes were consistent with the results of using the conventional real-time PCR, demonstrating a great potential of the MIP-enabled digital PCR in detecting gene expression in clinical samples.
数字 PCR 实现了稀有生物变体的高灵敏度和定量测量。本文引入了一种新的数字液滴式 PCR 技术，该技术通过使用具有一次性微流控芯片的微流控冲击打印机（MIP）将遗传靶标分成平面纳升级液滴阵列。通过检测跨越四个数量级（从 0.464 拷贝/μL 到 464 拷贝/μL）的 GAPDH 基因的一系列浓度梯度，验证了这种 MIP 增强型 PCR 技术的准确性。此外，该技术还应用于检测结肠癌组织和相邻非肿瘤组织中 p53 基因的表达，从中可以绝对定量确定核酸的拷贝数。结果与使用常规实时 PCR 的结果一致，表明 MIP 增强型数字 PCR 在检测临床样本中的基因表达方面具有很大的潜力。
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