恒温热隔绝式PCR技术的原理与应用  

Principle and Application of Insulated Isothermal PCR

作　　者：王娟[1] 卞国志 王贵平[1] 袁建丰[1] 

机构地区：[1]广东海洋大学畜牧兽医研究院有限公司,广东广州511400 [2]华南农业大学兽医学院,广东广州510642

出　　处：《动物医学进展》2017年第12期114-118,共5页Progress In Veterinary Medicine

基　　金：广东省科技计划项目(2014A020208052);广州市科技计划项目(201510010250)

摘　　要：由于常规PCR所使用的加热机制为热传导的热循环仪,需要一个精密的数字控制器来快速加热和冷却样品。近年来,越来越多的研究致力于开发一种快速、便携式的核酸检测PCR平台。而自然对流作为另一种热转换方式,能促进液体自发反复循环以提供工作所需的变性、退火和延伸温度,在核酸检测领域有很大的应用前景。恒温热隔绝式PCR(insulated isothermal PCR,iiPCR)就是采用Rayleigh-Be′nard自然对流原理,利用仪器底部单一热源对特殊聚碳酸酯毛细反应管底部的紫铜圈接触加热,毛细管上部置入隔热挡板,上下的温度差从而使毛细管内部流体对流自发形成可供PCR扩增反应的连续的温度梯度,LED屏上直接显示检测结果。与传统的PCR检测方法相比,具有操作简便、快速高效、特异性强、仪器成本低等优点,已开始运用于临床各种病原体的检测。Because the heating mechanism used in convention PCR based on thermal conductivity still require a delicate digital controller to rapidly heat and cool the sample,in recent years,a growing number of studies have focused on developing a fast and portable PCR platform for nucleic acid testing.Natural convection,another mode of heat transfer, promotes spontaneous and repeated liquid circulation to provide working temperatures for denaturation,annealing and extension.Insulated isothermal PCR （iiPCR）, a rapid nucleic acid detection method, based on Rayleigh-Be＇nard convection principle,amplification is carried in an iiPCR device in which a special polyearbonate capillary tube（R-tubeTM） is heated isothermally by a copper ring attached to tis bottom and shielded by a thermal baffle around its upper half, spontaneous fluid convection in a R-tube is driven by temperature gradients for PCR reaction.The results show directly on LED display.Compared with the traditional PCR; iiPCR assay shows its advantages in the rapid detection of clinical nucleic acid, and has begun to use in the field of clinical diagnosis of various pathogens for its low cost, simple and easy operation, high specificity, high sensibility and high amplification efficiency. Here we proceeded a review for insulated isothermal PCR covering its introduction, research progress and application prospect.

关 键 词：聚合酶链反应 瑞利-本纳德对流 恒温热隔绝式PCR 

分 类 号：S852.4[农业科学—基础兽医学]