MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou 510631, China.
Lab Chip. 2017 Dec 5;17(24):4334-4341. doi: 10.1039/c7lc01107e.
In this work, we describe for the first time a genetic diagnosis platform employing a polydiallyldimethylammonium chloride (PDDA)-modified capillary and a liquid-based thermalization system for rapid, simple and quantitative DNA analysis with minimal user interaction. Positively charged PDDA is modified on the inner surface of the silicon dioxide capillary by using an electrostatic self-assembly approach that allows the negatively charged DNA to be separated from the lysate in less than 20 seconds. The capillary loaded with the PCR mix is incorporated in the thermalization system, which can achieve on-site real-time PCR. This system is based on the circulation of pre-heated liquids in the chamber, allowing for high-speed thermalization of the capillary and fast amplification. Multiple targets can be simultaneously analysed with multiplex spatial melting. Starting with live Escherichia coli (E. coli) cells in milk, as a realistic sample, the current method can achieve DNA extraction, amplification, and detection within 40 min.
在这项工作中，我们首次描述了一种遗传诊断平台，该平台采用聚二烯丙基二甲基氯化铵 (PDDA) 修饰的毛细管和基于液体的热平衡系统，可实现快速、简单和定量的 DNA 分析，用户交互最少。带正电荷的 PDDA 通过静电自组装方法修饰在二氧化硅毛细管的内表面上，从而使带负电荷的 DNA 在不到 20 秒的时间内从裂解液中分离出来。将加载有 PCR 混合物的毛细管装入热平衡系统中，该系统可实现现场实时 PCR。该系统基于腔室内预加热液体的循环，允许毛细管快速热平衡和快速扩增。通过多重空间熔融，可以同时分析多个目标。从牛奶中的活大肠杆菌 (E. coli) 细胞开始，作为真实样本，该方法可以在 40 分钟内实现 DNA 提取、扩增和检测。
