微流控振荡流反转录聚合酶链式反应快速检测烟草花叶病毒  

Rapid Detection of Tobacco Mosaic Virus from Crude Samples on Oscillatory-flow Reverse Transcription Polymerase Chain Reaction Microfluidics

作　　者：王海英[1] 章春笋[1] 李彧媛[1] 

机构地区：[1]华南师范大学激光生命科学教育部重点实验室暨激光生命科学研究所,广州510631

出　　处：《分析化学》2009年第9期1286-1290,共5页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金(Nos.30700155;30870676;30800261);国家高技术研究发展计划(863计划;No.2007AA10Z204);教育部"长江学者与创新团队计划"创新团队项目(No.IRT0829)资助

摘　　要：建立了一种基于毛细管的振荡流反转录聚合酶链式反应(RT-PCR)的微流控装置检测烟草花叶病毒(TMV)的新方法。根据TMV移动蛋白基因设计一对PCR引物,对粗提纯TMV颗粒直接进行RT-PCR。用0.025%小牛血清蛋白(BSA)对反应毛细管内壁进行静力学和动力学钝化,提高了PCR效率。在此微流控装置上进行RT-PCR,流速为70μL/min时,检出限为0.01μg cDNA;可以在17min内成功扩增出179bp的目的DNA片段。An oscillatory-flow reverse transcription-polymerase chain reaction（RT-PCR） microfluidics based on a capillary was developed for the detection of tobacco mosaic virus（TMV）.A pair of PCR primers was designed according to the TMV movement protein gene.Direct RT-PCR-amplified products,a 179 base pair（bp） DNA fragment,from TMV particles were obtained.0.025% bovine serum albumin was used for the static and dynamic passivation to improve the PCR efficiency.On this device,the detection limit of 0.01 μg cDNA was achieved at the flow rate of 70 μL/min,and the amplification of 179 bp fragments can be successfully performed within 17 min.

关 键 词：微流控 振荡流 反转录聚合酶链式反应 烟草花叶病毒 

分 类 号：S432.41[农业科学—植物病理学]