毛细管液滴中的乙型肝炎病毒DNA提取与扩增  

In-capillary droplet-based hepatitis virus B DNA extraction and amplification

作　　者：王维[1] 刘大渔[1] 梁广铁[1] 雷秀霞[1] 周小棉[1] 

机构地区：[1]广州医学院附属广州市第一人民医院临床化学技术研究室,510180

出　　处：《国际检验医学杂志》2011年第13期1410-1413,共4页International Journal of Laboratory Medicine

基　　金：国家自然科学基金资助项目(30870753);科技部国际合作项目(2010DFB3380);广东省自然科学基金资助项目(9151008901000190);广东省医学科研基金资助项目(A2009518;A2010458);广州市医药卫生科技重点项目(2009-ZDi-10;2009-ZDi-21);广州市科信局科技支撑计划资助项目(2010J-E241-1)

摘　　要：目的利用毛细管液滴技术,发展一种集成DNA提取与扩增的乙型肝炎病毒分析方法。方法利用聚四氟乙烯毛细管的疏水特性,向毛细管中先后引入油相和水相溶液,在表面张力作用下形成油包水液滴。顺序向毛细管中引入含有不同试样的液滴,完成进样、DNA结合、洗涤、洗脱,扩增等过程。微液滴不仅有效解决了样品蒸发和扩散问题,同时在操作中起到微阀和微混合器微功能。结果利用上述毛细管液滴方法分析了乙型肝炎病毒的血清样品,通过优化DNA提取和扩增条件,毛细管液滴方法能在较短时间内高效纯化并扩增DNA。结论上述方法具有简便快速和低成本的优势,有望发展成一种有潜力的微全核酸分析系统。Objective To develop a droplet-based method for in-capillary hepatitis virus B DNA extraction and amplification.Methods The water-in-oil droplets were formed in the capillary as the result of the hydrophobic nature of PTFE capillary.By serially introducing droplets containing sample or reagents,DNA Binding,magnetic beads rinsing,DNA elution and PCR were performed in the capillary.The droplets served as micro-valve,micro-mixer and micro-reactor during the operation.The in-capillary droplet-based method was used to analyze hepatitis virus B containing serum samples.Results Experimental results showed that DNA extraction,purification and PCR amplification can be effectively coupled in the capillary.Conclusion The current method is simple,fast and low-cost,thus has the potential to be further developed into a micro-total nucleic acid analysis system.

关 键 词：聚合酶链反应 微全核酸分析系统 DNA提取 液滴 毛细管 

分 类 号：R440[医药卫生—诊断学]