One-step pathogen specific DNA extraction from whole blood on a centrifugal microfluidic device
One-step pathogen specific DNA extraction from whole blood on a centrifugal microfluidic device
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DOI：
10.1039/b616115d
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发表时间：
2007-01-01
期刊：
影响因子：
6.1
通讯作者：
Ko, Christopher
中科院分区：
文献类型：
--
作者：
Cho, Yoon-Kyoung;Lee, Jeong-Gun;Ko, Christopher
我们报告了一种完全集成的，病原体特异性DNA提取设备，利用基于聚合物的CD平台上的离心式微流控技术。通过使用创新的激光辐照铁蜡微阀(LIFM)和使用激光照射磁性颗粒的快速细胞裂解方法，我们可以首次在CD上展示完全集成的病原体特异性DNA提取方法。作为模型研究，进行了乙肝病毒(乙肝病毒)和大肠杆菌掺杂的全血DNA提取实验。血浆分离、与靶向特异性抗体偶联的磁珠混合、血浆残留去除、洗涤和DNA提取的全过程在12分钟内完成，仅需一个手动步骤，装载100亩L全血。实时定量聚合酶链式反应结果表明，用便携式样品制备仪在CD上制备的DNA浓度与传统的台式制样方法一样好。这表明，我们的新型离心式微流控平台能够完全集成需要多步骤流体控制的复杂生物反应。
We report a fully integrated, pathogen-specific DNA extraction device utilizing centrifugal microfluidics on a polymer based CD platform. By use of the innovative laser irradiated Ferrowax microvalve (LIFM) together with the rapid cell lysis method using laser irradiation on magnetic particles, we could, for the first time, demonstrate a fully integrated pathogen specific DNA extraction from whole blood on a CD. As a model study, DNA extraction experiments from whole blood spiked with Hepatitis B virus (HBV) and E. coli were conducted. The total process of the plasma separation, mixing with magnetic beads conjugated with target specific antibodies, removal of plasma residual, washing and DNA extraction was finished within 12 min with only one manual step, the loading of 100 mu L of whole blood. Real-time PCR results showed that the concentration of DNA prepared on a CD using a portable sample preparation device was as good as those by conventional bench top protocol. It demonstrates that our novel centrifugal microfluidics platform enables a full integration of complex biological reactions that require multi-step fluidic control.