本文關(guān)鍵詞：微流控對流PCR系統(tǒng)的研究與實現(xiàn)　出處：《北京化工大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 分子診斷 對流PCR 微流控 溫度控制 熒光檢測

【摘要】：基因擴增分子診斷技術(shù)具有靈敏度高、特異性好、技術(shù)成熟等特點,被廣泛應(yīng)用于體外診斷領(lǐng)域。PCR(Polymerase Chain Reaction)技術(shù)自1985年發(fā)明以來,研究者不斷以減少熱循環(huán)所需的反應(yīng)時間、降低儀器復(fù)雜度、簡化檢測操作等為目標進行改進優(yōu)化。本課題研究了一種基于微流控毛細管自環(huán)流原理的新型熒光定量對流PCR檢測方法與系統(tǒng)。此方法通過在微流控毛細管內(nèi)形成試劑的熱環(huán)流實現(xiàn)熱循環(huán),顯著縮短了傳統(tǒng)PCR熱循環(huán)所需時間；無需外界流體驅(qū)動系統(tǒng),降低了儀器復(fù)雜度；基于恒溫控制避免往復(fù)升降溫,減小了溫控系統(tǒng)的復(fù)雜度；通過設(shè)計了一種低成本、高靈敏度的熒光檢測方法,實現(xiàn)了快速檢測,縮短了檢測時間。本文首先介紹了實時毛細管對流PCR基因擴增系統(tǒng)的工作原理,然后分別介紹了系統(tǒng)總體設(shè)計、熒光檢測模塊設(shè)計和溫度控制模塊設(shè)計與實現(xiàn)。在儀器設(shè)計方面,本課題采用了雙處理器架構(gòu),實現(xiàn)了人機交互功能與系統(tǒng)功能的相互配合；在熒光檢測方面,本課題基于光纖、透鏡、濾光片等光學(xué)器件設(shè)計了光學(xué)子系統(tǒng),并基于LED、光電二極管等器件設(shè)計了一種成本低、結(jié)構(gòu)緊湊的熒光檢測模塊來實現(xiàn)高靈敏度實時熒光檢測；在溫度控制方面,本課題引入了基于內(nèi)模控制的PID整定方法,實現(xiàn)了快速整定溫度控制系統(tǒng)的控制器參數(shù)。本課題構(gòu)建了一種結(jié)構(gòu)簡單、成本低廉、使用方便的快速核酸擴增檢測系統(tǒng),在30分鐘內(nèi)即可完成基于核酸擴增的分子診斷,并通過對H1N1流感病毒實現(xiàn)成功檢測,驗證了此系統(tǒng)綜合性能。
[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.
【學(xué)位授予單位】：北京化工大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：TH776

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本文編號：1426816