School of Integrative Engineering, Chung-Ang University, Heuksuk-dong, Dongjak-gu, Seoul 156-756, Republic of Korea.
School of Integrative Engineering, Chung-Ang University, Heuksuk-dong, Dongjak-gu, Seoul 156-756, Republic of Korea.
Biosens Bioelectron. 2015 May 15;67:625-33. doi: 10.1016/j.bios.2014.09.083. Epub 2014 Oct 5.
Sample preparation has recently been an issue in the detection of food poisoning pathogens, particularly viruses such as norovirus (NoV), in food because of the complexity of foods and raw fresh materials. Here, we demonstrate a total analytical microfluidic chip module to automatically perform a series of essential processes (cell concentration, lysis (RNA extraction), nucleic acid amplification, and detection) for the fast but sensitive detection of norovirus in oysters. The murine NoV spiked oyster was stomached using a standard method. The supernatant was first loaded into a shape switchable sample preparation chamber consisting of charge switchable micro-beads. Murine NoV, which was adsorbed on microbeads by electrostatic physisorption, was lysed using bead beating. The extracted RNA was transferred to the detection chamber to be amplified using Nucleic Acid Sequence Based Amplification (NASBA). The optimal surface functionality, size, and number of microbeads were achieved for the virus concentration and the stable RNA extraction in the shape-switchable micro-channel. As a result, murine NoV in a single oyster was successfully detected within 4h by the microfluidic chip developed here, and could be directly applied to the large volume environmental sample as well as the food sample.
样品制备最近一直是食品中毒性病原体检测的一个问题，特别是病毒，如诺如病毒（NoV），因为食品和原材料的复杂性。在这里，我们展示了一个总分析微流控芯片模块，用于自动执行一系列基本过程（细胞浓缩、裂解（RNA 提取）、核酸扩增和检测），用于快速但敏感地检测牡蛎中的诺如病毒。使用标准方法对含有鼠诺如病毒的牡蛎进行胃灌。首先将上清液加载到由电荷可切换微珠组成的形状可切换样品制备室中。通过静电物理吸附吸附在微珠上的鼠诺如病毒通过珠粒破碎裂解。提取的 RNA 被转移到检测室中，使用基于核酸序列的扩增（NASBA）进行扩增。在形状可切换微通道中，实现了最佳的表面功能、微珠的大小和数量，以实现病毒浓缩和稳定的 RNA 提取。结果，通过这里开发的微流控芯片，成功地在单个牡蛎中检测到了鼠诺如病毒，并且可以直接应用于大容量的环境样品以及食品样品。
