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开发一种非传输微滴生成芯片和数字分子检测系统

Rui Sun^1,2, Lei Li^1,2, Tong Fang³

¹School of Biomedical Engineering (Suzhou), Division of Life Sciences and Medicine, University of Science and Technology of China, Suzhou, Jiangsu 215163, P. R. China.

Analytical methods : advancing methods and applications

|May 29, 2025

概括

一个新型的微滴生成芯片使得封闭工作流数字PCR (ddPCR) 能够进行敏感的核酸检测. 这种具有成本效益的系统简化了工作流程,并提高了液体活检等应用程序的准确性.

科学领域:

生物技术是生物技术.
分子生物学分子生物学
分析化学分析化学

背景情况:

滴滴数字PCR (ddPCR) 是核酸量化的一种强大技术.
现有的ddPCR方法通常涉及手动转移步骤,增加污染风险和复杂性.
需要简化,具有高灵敏度和可重复性,具有成本效益的ddPCR平台.

研究的目的:

开发一种用于集成ddPCR的新型无转移微滴生成芯片.
为了评估新的基于芯片的ddPCR系统的性能.
在成本,工作流程和准确性方面展示开发系统的优势.

主要方法:

微流体芯片的制造,使用交叉连接流量聚焦机制来产生滴滴.
在芯片上的封闭系统中集成滴滴生成和PCR放大.
开发一个定制的光阅读器用于滴滴分析.
使用ALB4DNA标准对商用ddPCR系统 (Bio-Rad QX200) 的比较性能分析.

主要成果:

该芯片成功地产生了均大小的油中的水滴.
综合系统实现了1-10^4副本/μL的动态范围,具有高可重复性 (CV <10%).
准确性和线性与商业系统相比 (回收率:95-129%与95-131%,R^2 = 0.998与0.999).
确定了关键优势:成本效益,消除转移步骤和简化工作流.

结论:

开发的无转移微滴生成芯片可以实现实用和高效的ddPCR工作流.
该系统的准确性与现有平台相提并论,但具有显著的成本和工作流的好处.
这项创新适用于在液体活检和病原体监测等应用中敏感核酸检测.

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