Mamaev D D, Khodakov D A, Dement'eva E I, Filatov I V, Iurasov D A, Cherepanov A I, Vasiliskov V A, Smoldovskaia O V, Zimenkov D V, Griadunov D A, Mikhaĭlovich V M, Zasedatelev A S
Prikl Biokhim Mikrobiol. 2011 Mar-Apr;47(2):231-40.
A method and a microfluidic device for automated extraction and purification of nucleic acids from biological samples have been developed. The method involves disruption of bacterial cells and/or viral particles by combining enzymatic and chemical lysis procedures followed by solid-phase sorbent extraction and purification of nucleic acids. The procedure is carried out in an automated mode in a microfluidic module isolated from the outside environment, which minimizes contact of the researcher with potentially infectious samples and, consequently, decreases the risk of laboratory-acquired infections. The module includes reservoirs with lyophilized components for lysis and washing buffers; a microcolumn with a solid-phase sorbent; reservoirs containing water, ethanol, and water-ethanol buffer solutions for dissolving freeze-dried buffer components, rinsing the microcolumn, and eluting of nucleic acids; and microchannels and valves needed for directing fluids inside the module. The microfluidic module is placed into the control unit that delivers pressure, heats, mixes reagents, and flows solutions within the microfluidic module. The microfluidic system performs extraction and purification of nucleic acids with high efficiency in 40 min, and nucleic acids extracted can be directly used in PCR reaction and microarray assays.
已开发出一种用于从生物样品中自动提取和纯化核酸的方法及微流控装置。该方法包括通过结合酶解和化学裂解程序来破坏细菌细胞和/或病毒颗粒，随后进行固相吸附剂提取和核酸纯化。该程序在与外部环境隔离的微流控模块中以自动化模式进行，这最大限度地减少了研究人员与潜在感染性样品的接触，从而降低了实验室获得性感染的风险。该模块包括装有用于裂解和洗涤缓冲液的冻干成分的储液器；带有固相吸附剂的微柱；装有用于溶解冻干缓冲液成分、冲洗微柱和洗脱核酸的水、乙醇和水 - 乙醇缓冲溶液的储液器；以及用于在模块内引导流体的微通道和阀门。微流控模块放置在控制单元中，该控制单元在微流控模块内提供压力、加热、混合试剂并使溶液流动。微流控系统在40分钟内高效地进行核酸的提取和纯化，提取的核酸可直接用于PCR反应和微阵列分析。
