恒温热隔绝式PCR技术的原理与应用

发布时间：2018-12-24 13:54

【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。
[Abstract]:Because the heating mechanism used in conventional PCR is a heat conduction thermometer, a precise digital controller is needed to quickly heat and cool samples. In recent years, more and more research has been devoted to the development of a fast, portable PCR platform for nucleic acid detection. As another way of heat transfer, natural convection can promote the spontaneous repeated circulation of liquid to provide the necessary denaturation, annealing and extension temperature for work. It has great application prospect in the field of nucleic acid detection. Constant temperature and heat isolation type PCR (insulated isothermal PCR,iiPCR) is based on the principle of Rayleigh-Be'nard natural convection, using a single heat source at the bottom of the instrument to heat the copper ring at the bottom of the special polycarbonate capillary tube, and placing the upper part of the capillary tube into an insulating baffle. The temperature difference between the upper and lower levels makes the convection in the capillary tube spontaneously form a continuous temperature gradient for PCR amplification reaction. The detection results are directly displayed on the LED screen. Compared with traditional PCR detection method, it has many advantages, such as simple operation, high efficiency, specificity and low cost of instrument, so it has been applied to the detection of clinical pathogens.
【作者单位】：广东海洋大学畜牧兽医研究院有限公司;华南农业大学兽医学院;
【基金】：广东省科技计划项目(2014A020208052) 广州市科技计划项目(201510010250)
【分类号】：S854.4

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