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微流控对流PCR系统的研究与实现

发布时间:2018-01-15 04:31

  本文关键词:微流控对流PCR系统的研究与实现 出处:《北京化工大学》2016年硕士论文 论文类型:学位论文


  更多相关文章: 分子诊断 对流PCR 微流控 温度控制 荧光检测


【摘要】:基因扩增分子诊断技术具有灵敏度高、特异性好、技术成熟等特点,被广泛应用于体外诊断领域。PCR(Polymerase Chain Reaction)技术自1985年发明以来,研究者不断以减少热循环所需的反应时间、降低仪器复杂度、简化检测操作等为目标进行改进优化。本课题研究了一种基于微流控毛细管自环流原理的新型荧光定量对流PCR检测方法与系统。此方法通过在微流控毛细管内形成试剂的热环流实现热循环,显著缩短了传统PCR热循环所需时间;无需外界流体驱动系统,降低了仪器复杂度;基于恒温控制避免往复升降温,减小了温控系统的复杂度;通过设计了一种低成本、高灵敏度的荧光检测方法,实现了快速检测,缩短了检测时间。本文首先介绍了实时毛细管对流PCR基因扩增系统的工作原理,然后分别介绍了系统总体设计、荧光检测模块设计和温度控制模块设计与实现。在仪器设计方面,本课题采用了双处理器架构,实现了人机交互功能与系统功能的相互配合;在荧光检测方面,本课题基于光纤、透镜、滤光片等光学器件设计了光学子系统,并基于LED、光电二极管等器件设计了一种成本低、结构紧凑的荧光检测模块来实现高灵敏度实时荧光检测;在温度控制方面,本课题引入了基于内模控制的PID整定方法,实现了快速整定温度控制系统的控制器参数。本课题构建了一种结构简单、成本低廉、使用方便的快速核酸扩增检测系统,在30分钟内即可完成基于核酸扩增的分子诊断,并通过对H1N1流感病毒实现成功检测,验证了此系统综合性能。
[Abstract]:The molecular diagnostic technique of gene amplification has the characteristics of high sensitivity, good specificity and mature technology. Polymerase Chain reaction has been widely used in the field of in vitro diagnosis since its invention in 1985. Researchers continue to reduce the reaction time required for the thermal cycle and reduce the complexity of the instrument. A new fluorescent quantitative convection PCR detection method and system based on the principle of self-circulation of microfluidic capillary is studied. The thermal circulation of the inner forming reagent realizes the thermal cycle. The time required for the traditional PCR thermal cycle is shortened significantly. The complexity of the instrument is reduced without the need of the external fluid drive system. Based on constant temperature control, the complexity of temperature control system is reduced. By designing a low cost and high sensitivity fluorescence detection method, the rapid detection is achieved and the detection time is shortened. Firstly, the principle of real-time capillary convection PCR amplification system is introduced in this paper. Then the overall design of the system, fluorescence detection module design and temperature control module design and implementation. In the instrument design, the subject uses a dual-processor architecture. The man-machine interaction function and the system function cooperation are realized. In the aspect of fluorescence detection, optical subsystem is designed based on optical devices such as optical fiber, lens, filter and so on, and a kind of low cost is designed based on LED, photodiode and so on. A compact fluorescent detection module is used to realize real-time fluorescence detection with high sensitivity. In the aspect of temperature control, the PID tuning method based on internal model control is introduced in this paper, and the controller parameters of the temperature control system are quickly set up. A simple structure and low cost are constructed in this paper. Using a convenient rapid nucleic acid amplification detection system, the molecular diagnosis based on nucleic acid amplification can be completed within 30 minutes, and the comprehensive performance of the system is verified by successfully detecting H1N1 influenza virus.
【学位授予单位】:北京化工大学
【学位级别】:硕士
【学位授予年份】:2016
【分类号】:TH776

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