Yang Sung-Yi, Lien Kang-Yi, Huang Kao-Jean, Lei Huan-Yao, Lee Gwo-Bin
Department of Engineering Science, National Cheng Kung University, 1 University Road, Tainan 701, Taiwan.
Biosens Bioelectron. 2008 Dec 1;24(4):861-8. doi: 10.1016/j.bios.2008.07.019. Epub 2008 Jul 22.
The current study presents a new miniature microfluidic flow cytometer integrated with several functional micro-devices capable of viral sample purification and detection by utilizing a magnetic bead-based immunoassay. The magnetic beads were conjugated with specific antibodies, which can recognize and capture target viruses. Another dye-labeled anti-virus antibody was then used to mark the bead-bound virus for the subsequent optical detection. Several essential components were integrated onto a single chip including a sample incubation module, a micro flow cytometry module and an optical detection module. The sample incubation module consisting of pneumatic micropumps and a membrane-type, active micromixer was used for purifying and enriching the target virus-bound magnetic beads with the aid of a permanent magnet. The micro flow cytometry module and the optical detection module were used to perform the functions of virus counting and collection. Experimental results showed that virus samples with a concentration of 10(3)PFU/ml can be automatically detected successfully by the developed system. In addition, the entire diagnosis procedure including sample incubation and virus detection took only about 40min. Consequently, the proposed micro flow cytometry may provide a powerful platform for rapid diagnosis and future biological applications.
当前的研究展示了一种新型微型微流控流式细胞仪，它集成了多种功能微器件，能够利用基于磁珠的免疫测定法对病毒样本进行纯化和检测。磁珠与特定抗体结合，这些抗体能够识别并捕获目标病毒。然后使用另一种染料标记的抗病毒抗体标记与磁珠结合的病毒，以便进行后续的光学检测。几个关键组件集成在一个芯片上，包括样品孵育模块、微流控细胞仪模块和光学检测模块。由气动微型泵和膜式有源微混合器组成的样品孵育模块，借助永磁体用于纯化和富集与目标病毒结合的磁珠。微流控细胞仪模块和光学检测模块用于执行病毒计数和收集功能。实验结果表明，所开发的系统能够成功自动检测浓度为10(3)PFU/ml的病毒样本。此外，包括样品孵育和病毒检测在内的整个诊断过程仅需约40分钟。因此，所提出的微流控细胞仪可能为快速诊断及未来的生物应用提供一个强大的平台。
