聚合酶链式反应(PCR)是一种在体外实现特定基因片段复制的生物学技术,目前已广泛应用于病原体诊断,基因突变检测,食品科学等领域。然而,传统的PCR热循环仪不仅体积大,而且热转换效率相对较低,一般为4℃/s~6℃/s,导致基因扩增时间相对较...聚合酶链式反应(PCR)是一种在体外实现特定基因片段复制的生物学技术,目前已广泛应用于病原体诊断,基因突变检测,食品科学等领域。然而,传统的PCR热循环仪不仅体积大,而且热转换效率相对较低,一般为4℃/s~6℃/s,导致基因扩增时间相对较长。微流控芯片又称“芯片实验室”,它是将多个不同功能的单元集成在一起以实现样品进样、化学反应及检测等。本文旨在介绍以微流控芯片为基础的自然对流与连续流技术的快速PCR研究方面的进展。Polymerase chain reaction (PCR) is a biological technology to replicate specific gene fragments in vitro, which has been widely used in pathogen diagnosis, gene mutation detection, food science and other fields. However, the traditional PCR thermal cycling instrument is not only large in size, but also relatively low in thermal conversion efficiency. Generally, the temperature variation rate is about 4˚C/s~6˚C/s, resulting in relatively long gene amplification time. Microfluidic chip, also known as “Lab-on-a-Chip”, integrates multiple units with different functions to achieve sample injection, chemical reaction and detection. The purpose of this paper is to introduce the development of natural convection and continuous flow PCR based on microfluidic chips.展开更多
一种高通量振荡流PCR微流控技术已被成功开发,借此来快速检测食品致病菌-单增李斯特氏菌(L.monocytogenes)。该PCR微流控装置主要由基于LabView的温度控制与采集系统、三个铜加热块以及聚四氟乙烯(PTFE)毛细管等构成。在该微流控装置上...一种高通量振荡流PCR微流控技术已被成功开发,借此来快速检测食品致病菌-单增李斯特氏菌(L.monocytogenes)。该PCR微流控装置主要由基于LabView的温度控制与采集系统、三个铜加热块以及聚四氟乙烯(PTFE)毛细管等构成。在该微流控装置上,三个铜块维持PCR反应所需要的三个温度,而包埋在铜块沟槽中的PTFE毛细管作为PCR的反应通道。毛细管通道中的PCR溶液通过精密注射泵驱动实现往复振荡,从而实现基于振荡流PCR的DNA快速扩增。当流速为300μL/min时,135 bp DNA基因片段能在14min内成功扩增,L.monocytogenes检测极限为10 cfu/mL。目前开发的高通量振荡流PCR系统能同时检测水、牛奶、香蕉以及香肠中的L.monocytogenes。展开更多
A novel continuous-flow PCR chip adopting self-heating, passive-cooling mode to realize the DNA fragments amplification was presented. Using the ANSYS finite element analysis, the temperature distribution of the chip ...A novel continuous-flow PCR chip adopting self-heating, passive-cooling mode to realize the DNA fragments amplification was presented. Using the ANSYS finite element analysis, the temperature distribution of the chip is simulated and analyzed.The optimal size of the chip is 30×22 mm2, the roundabout micro-channel is the 90 μm width, 40 μm depth. Two micro-heater with the nickel-chrome alloy material film are formed on the side of silicon belonging to denaturation and renaturation zones needed for PCR reaction, and two adiabatic structures with groove on side of silicon by anisotropy etching. By the mode of heating local zones at single side, three wider constant temperature zones could be formed, which are 60 ℃,72 ℃,95 ℃ and suitable for PCR,and the temperature-difference could be restricted in less than 5 ℃.展开更多
文摘聚合酶链式反应(PCR)是一种在体外实现特定基因片段复制的生物学技术,目前已广泛应用于病原体诊断,基因突变检测,食品科学等领域。然而,传统的PCR热循环仪不仅体积大,而且热转换效率相对较低,一般为4℃/s~6℃/s,导致基因扩增时间相对较长。微流控芯片又称“芯片实验室”,它是将多个不同功能的单元集成在一起以实现样品进样、化学反应及检测等。本文旨在介绍以微流控芯片为基础的自然对流与连续流技术的快速PCR研究方面的进展。Polymerase chain reaction (PCR) is a biological technology to replicate specific gene fragments in vitro, which has been widely used in pathogen diagnosis, gene mutation detection, food science and other fields. However, the traditional PCR thermal cycling instrument is not only large in size, but also relatively low in thermal conversion efficiency. Generally, the temperature variation rate is about 4˚C/s~6˚C/s, resulting in relatively long gene amplification time. Microfluidic chip, also known as “Lab-on-a-Chip”, integrates multiple units with different functions to achieve sample injection, chemical reaction and detection. The purpose of this paper is to introduce the development of natural convection and continuous flow PCR based on microfluidic chips.
文摘一种高通量振荡流PCR微流控技术已被成功开发,借此来快速检测食品致病菌-单增李斯特氏菌(L.monocytogenes)。该PCR微流控装置主要由基于LabView的温度控制与采集系统、三个铜加热块以及聚四氟乙烯(PTFE)毛细管等构成。在该微流控装置上,三个铜块维持PCR反应所需要的三个温度,而包埋在铜块沟槽中的PTFE毛细管作为PCR的反应通道。毛细管通道中的PCR溶液通过精密注射泵驱动实现往复振荡,从而实现基于振荡流PCR的DNA快速扩增。当流速为300μL/min时,135 bp DNA基因片段能在14min内成功扩增,L.monocytogenes检测极限为10 cfu/mL。目前开发的高通量振荡流PCR系统能同时检测水、牛奶、香蕉以及香肠中的L.monocytogenes。
基金the National Natural Science Foundation of China(Grant No.60576047)
文摘A novel continuous-flow PCR chip adopting self-heating, passive-cooling mode to realize the DNA fragments amplification was presented. Using the ANSYS finite element analysis, the temperature distribution of the chip is simulated and analyzed.The optimal size of the chip is 30×22 mm2, the roundabout micro-channel is the 90 μm width, 40 μm depth. Two micro-heater with the nickel-chrome alloy material film are formed on the side of silicon belonging to denaturation and renaturation zones needed for PCR reaction, and two adiabatic structures with groove on side of silicon by anisotropy etching. By the mode of heating local zones at single side, three wider constant temperature zones could be formed, which are 60 ℃,72 ℃,95 ℃ and suitable for PCR,and the temperature-difference could be restricted in less than 5 ℃.
