On-line cell lysis and DNA extraction on a microfluidic biochip fabricated by microelectromechanical system technology
On-line cell lysis and DNA extraction on a microfluidic biochip fabricated by microelectromechanical system technology
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DOI：
10.1002/elps.200700551
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发表时间：
2008-05-01
期刊：
影响因子：
2.9
通讯作者：
Liu, Chang Chun
中科院分区：
文献类型：
--
作者：
Chen, Xing;Cui, Da Fu;Liu, Chang Chun
将细胞裂解和DNA纯化过程集成到微全分析系统（mu TAS）中是核酸分析的关键步骤之一。基于化学方法的芯片上细胞裂解通过血液样品和裂解试剂的充分混合来实现。本文提出了两种混合模型：T型混合模型和T型混合模型，并对这两种模型进行了模拟。模拟结果表明，螺旋通道的螺旋型混合模型的性能最好，该模型进一步用于构建在线细胞裂解和DNA提取的微流控生物芯片。仿真结果得到了实验的进一步验证。它声称血液样本和裂解试剂的混合率超过80%，这保证了当细胞/缓冲液速度比小于1：5时，可以在入口位置附近实现完全的细胞裂解。细胞裂解后，DNA提取通过固相法通过使用集成在生物芯片中的多孔硅基质来实现。在连续流动过程中的微芯片，快速细胞裂解和PCR扩增的基因组DNA纯化，可以在20分钟内实现。这种微流控生物芯片的潜力是通过预处理全血样品，这表明集成的可能性样品制备，PCR和分离在一个单一的设备上工作的便携式即时医疗诊断系统。
Integrating cell lysis and DNA purification process into a micrototal analytical system (mu TAS) is one critical step for the analysis of nucleic acids. On-chip cell lysis based on a chemical method is realized by sufficient blend of blood sample and the lyzing reagent. In this paper two mixing models, T-type mixing model and sandwich-type mixing model, are proposed and simulation of those models is conducted. Result of simulation shows that the sandwich-type mixing model with coiled channel performs best and this model is further used to construct the microfluidic biochip for on-line cell lysis and DNA extraction. The result of simulation is further verified by experiments. It asserts that more than 80% mixing of blood sample and lyzing reagent which guarantees that completed cell lysis can be achieved near the inlet location when the cell/buffer velocity ratio is less than 1:5. After cell lysis, DNA extraction by means of a solid-phase method is implemented by using porous silicon matrix which is integrated in the biochip. During continuous flow process in the microchip, rapid cell lysis and PCR-amplifiable genomic DNA purification can be achieved within 20 min. The potential of this microfluidic biochip is illustrated by pretreating a whole blood sample, which shows the possibility of integration of sample preparation, PCR, and separation on a single device to work as portable point-of-care medical diagnostic system.