基于微流控芯片巢氏等温扩增核酸检测仪设计

Design of Nested Isothermal Amplification Nucleic Acid Detector Based on Microfluidic Chip

针对呼吸道病原体检测需要依靠实验室内固定设备和专业技术人员,难以实现现场快速即时检测的问题和多种病原体同时检测的需求,开发了一种基于微流控芯片和重组聚合酶(RPA)等温扩增技术的便携式巢式等温扩增核酸检测仪。使用微流控芯片完成核酸纯化和扩增;模糊PID算法控制RPA需要的37℃等温扩增环境;读取扩增室的荧光值与判断阈值比较,定性检测核酸扩增结果;使用触摸屏进行人机交互操作,打印检测结果。通过对甲型H1N1流感病毒DNA质粒进行等温扩增和荧光检测,证明该仪器能够用于呼吸道病原核酸扩增检测,是一种理想的便携式现场核酸检测设备。

To solve the problem that the detection of respiratory pathogens depends on fixed equipment and professional technicians in the laboratory,which is difficult to achieve rapid and instant detection on site,and the need for simultaneous detection of multiple pathogens,a portable nested isothermal amplification nucleic acid detector based on microfluidic chip and recombinant polymerase(RPA)isothermal method is developed.The microfluidic chip is used to complete nucleic acid purification and amplification;the fuzzy PID algorithm controls the 37℃isothermal amplification environment required by RPA;the fluorescence value of the amplification chamber is readand compared with the judgment threshold to qualitatively detect the nucleic acid amplification results;the touch screen is used to perform the human-computer interactive operation and print the test result.Through isothermal amplification and fluorescence detection of influenza A HIN1 virus DNA plasmid,it is proved that the instrument can be used for the amplification and detection of respiratory pathogenic nucleic acid,and it is an ideal portable on-site nucleic acid detection equipment.