Ultra-rapid flow-through polymerase chain reaction microfluidics using vapor pressure
Ultra-rapid flow-through polymerase chain reaction microfluidics using vapor pressure
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DOI：
10.1016/j.bios.2011.06.022
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发表时间：
2011-09-15
影响因子：
12.6
通讯作者：
Tamiya, Eiichi
中科院分区：
文献类型：
--
作者：
Fuchiwaki, Yusuke;Nagai, Hidenori;Tamiya, Eiichi
开发了一种新型的蒸汽压流式聚合酶链反应（PCR）微流控系统，可以在芯片上实现超快速、小体积的DNA扩增。40个周期的扩增可以在短短120秒内完成，使该设备成为世界上最快的PCR系统。该芯片由压敏聚烯烃（PSP）薄膜和环烯烃聚合物（COP）衬底组成，通过切削加工制备微通道。该微通道的封闭结构仅通过在COP衬底上称重制成，具有良好的实际应用价值。利用不稳定流源变性区的蒸汽压作用于流动力，用最少的检测试剂实现了超快速、高效的扩增。蒸汽压产生的流动节奏使残留的PCR产物最小化，从而实现高效扩增。为了进行现场测试分析，从公共场所收集空气中的粉尘并检测炭疽菌的存在。该PCR芯片对炭疽菌鉴定具有足够的灵敏度。从气溶胶取样到检测的最快时间理论上估计为8分钟。(C) 2011 Elsevier B.V.版权所有
A novel flow-through polymerase chain reaction (PCR) microfluidic system using vapor pressure was developed that can achieve ultra-rapid, small-volume DNA amplification on a chip. The 40-cycle amplification can be completed in as little as 120s, making this device the fastest PCR system in the world. The chip device is made of a pressure-sensitive polyolefin (PSP) film and cyclo-olefin polymer (COP) substrate which was processed by cutting-work to fabricate the microchannel. The enclosed structure of the microchannel was fabricated solely by weighing the PSP film on the COP substrate, resulting in superior practical application. The vapor pressure in the denaturation zone of the destabilizing flow source was applied to the flow force, and ultra-rapid, efficient amplification was accomplished with a minimal amount of PCR reagents for detection. The flowing rhythm created by vapor pressure minimized the residual PCR products, leading to highly efficient amplification. For field test analysis, airborne dust was collected from a public place and tested for the presence of anthrax. The PCR chip had sufficient sensitivity for anthrax identification. The fastest time from aerosol sampling to detection was theoretically estimated as 8 min. (C) 2011 Elsevier B.V. All rights reserved.