Yeh Szu-I, Fang Wei-Feng, Huang Chao-Jyun, Wang Tzu-Ming, Yang Jing-Tang
Department of Mechanical Engineering, National Taiwan University.
Department of Mechanical Engineering, National Taiwan University;
J Vis Exp. 2016 Sep 27(115):54424. doi: 10.3791/54424.
A simple and visual method to detect multi-nucleotide polymorphism (MNP) was performed on a pneumatic droplet manipulation platform on an open surface. This approach to colorimetric DNA detection was based on the hybridization-mediated growth of gold nanoparticle probes (AuNP probes). The growth size and configuration of the AuNP are dominated by the number of DNA samples hybridized with the probes. Based on the specific size- and shape-dependent optical properties of the nanoparticles, the number of mismatches in a sample DNA fragment to the probes is able to be discriminated. The tests were conducted via droplets containing reagents and DNA samples respectively, and were transported and mixed on the pneumatic platform with the controlled pneumatic suction of the flexible PDMS-based superhydrophobic membrane. Droplets can be delivered simultaneously and precisely on an open-surface on the proposed pneumatic platform that is highly biocompatible with no side effect of DNA samples inside the droplets. Combining the two proposed methods, the multi-nucleotide polymorphism can be detected at sight on the pneumatic droplet manipulation platform; no additional instrument is required. The procedure from installing the droplets on the platform to the final result takes less than 5 min, much less than with existing methods. Moreover, this combined MNP detection approach requires a sample volume of only 10 µl in each operation, which is remarkably less than that of a macro system.
一种简单直观的检测多核苷酸多态性(MNP)的方法在开放表面的气动微滴操纵平台上进行。这种比色法DNA检测方法基于金纳米颗粒探针(AuNP探针)的杂交介导生长。AuNP的生长大小和形态由与探针杂交的DNA样本数量决定。基于纳米颗粒特定的尺寸和形状依赖性光学特性,能够区分样本DNA片段与探针之间错配的数量。测试分别通过含有试剂和DNA样本的微滴进行,并在基于柔性聚二甲基硅氧烷的超疏水膜的受控气动抽吸作用下在气动平台上运输和混合。微滴可以同时且精确地递送至所提出的气动平台的开放表面上,该平台具有高度生物相容性,对微滴内的DNA样本无副作用。结合所提出的两种方法,可以在气动微滴操纵平台上直观地检测多核苷酸多态性;无需额外的仪器。从将微滴安装在平台上到获得最终结果的过程耗时不到5分钟,远少于现有方法。此外,这种联合的MNP检测方法每次操作仅需要10微升的样本体积,这明显少于宏观系统所需的体积。