基于毛细管液滴技术的血液乙型肝炎病毒DNA提取  

Droplet-based Blood Hepatitis B Virus DNA Extraction in a Capillary

作　　者：梁广铁[1] 王秋平[1] 王维[1] 刘大渔[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检验科临床化学技术研究室,广州510180

出　　处：《分析化学》2011年第5期670-674,共5页Chinese Journal of Analytical Chemistry

基　　金："十一五"重大专项(No.2008ZX10004-004);国家自然科学基金(No.30870753);广州市医药卫生科技重点项目(No.2009-ZDi-21)资助

摘　　要：基于毛细管液滴技术,建立了提取血液中乙型肝炎病毒(HBV)DNA的方法。方法的基本原理是向聚四氟乙烯毛细管中连续引入油相和水相溶液时,由于表面张力作用,可以形成稳定的油包水型液滴。依次引入含有不同试样的液滴,在毛细管中完成进样、DNA结合、洗涤以及洗脱等过程。实验表明,采用蛋白酶K裂解和提高洗脱温度有利于提高DNA回收率。优化后的血清HBV DNA提取操作在25 min内完成,回收率大于60%。本方法的DNA提取量与常规方法相当,而相对标准偏差较大(23.3%vs 12.9%)。本方法具有简单、快速和低成本的优势,有望成为一种微全核酸分析系统样品前处理的有效解决方案。A novel in-capillary droplet-based DNA extraction method was developed,using the principle of water-in-oil droplet formation in the highly hydrophobic polytetrafluoroethylene capillary.By sequentially introducing droplets containing sample or reagents,all the steps related to blood hepatitis B virus（HBV） DNA extraction,including sample injection,DNA binding,magnetic beads rinsing and DNA elution were continuously implemented in the capillary.Experimental results showed that proteinase K lysis and elevated elution temperature facilitated higher extraction efficiency.With optimized conditions,an extraction was completed in 25 min with microliter level sample/reagent consumption.The droplet-based method achieved an extraction efficiency of ～60% that is comparable to that of a commercial method.However,the RSD of extraction efficiency was relatively higher.In summary,the droplet-based DNA extraction developed is simple,fast and low-cost.It has the potential to be integrated into a micro-total nucleic acid analysis system.

关 键 词：微全核酸分析系统 DNA提取 液滴 毛细管 乙型肝炎病毒 

分 类 号：R373.21[医药卫生—病原生物学]