实时PCR仪荧光信号监测系统的研究
详细信息    本馆镜像全文|  推荐本文 |  |   获取CNKI官网全文
摘要
聚合酶链反应(PCR)是生物医学领域的一项革命性创举,实时PCR技术实现了PCR从定性到定量的飞跃。实时PCR仪已经广泛应用于分子生物学和医学等领域,代表着PCR技术的发展趋势。实时PCR仪荧光信号监测系统是实时PCR仪的一个核心子系统,提供了对荧光信号的数据处理与监测,是定量分析的前提与基础。
     本文开发实时PCR仪荧光信号监测系统,通过实现对检测装置的运动控制、荧光信号的数据处理实现对荧光信号的监测,本文的主要内容如下:
     第一章阐述实时PCR系统在国内外的发展概况,阐述本文的研究目标和意义,提出本文的研究内容和方法。
     第二章介绍实时PCR仪荧光信号监测系统的硬件设计方案,阐述硬件结构的特点及优点,根据硬件设计方案,搭建硬件系统。
     第三章介绍实时PCR仪荧光信号监测系统的软件开发工具,从运动控制模块及荧光信号数据采集与处理模块两大方面对软件系统进行分析与设计。
     第四章介绍运动控制模块的实现方法,阐述插补技术的原理及分类,对数据采样插补算法进行优化并应用到本系统中,研究中断控制方法,对检测路径进行规划,最终实现对运动控制模块的控制。
     第五章介绍荧光信号数据采集与处理模块的实现方法,将多线程技术应用于数据采集与处理模块中,分别从数据段选取、数字滤波、数据平滑、峰值识别、信号归一化五个方面对荧光信号进行数据处理,最终实现对荧光信号的监测,通过仿真与实验相结合检验荧光信号监测系统性能。
     第六章对全文进行总结,对后续工作进行展望。
Polymerase chain reaction(PCR) is a revolutionary creation in the biomedical field, real-time PCR technology realizes a leap from qualitative to quantitative. real-time PCR instrument has been widely used in molecular biology and medicine, what's more, it's the development trend of PCR technology. As a core subsystem of real-time PCR instrument, fluorescence signal monitoring system is the precondition and foundation for quantitative analysis, it provides fluorescence signal data processing and monitoring.
     The fluorescence signal monitoring system of real-time PCR instrument has been developed in this paper, which could implement the motion control of detection device, the data processing and the monitoring of fluorescence signal. The following are the main contents of this paper:
     The first chapter discusses the development of real-time PCP instrument at home and abroad, explains research objectives and research significance, proposes research contents and research methods.
     The second chapter describes the hardware designing of fluorescence signal monitoring system, and establishes a hardware system according to the hardware designing.
     The third chapter describes the development tools of fluorescence signal monitoring software system, analyses and designs the software of motion control module and data acquisition and processing module.
     The forth chapter describes the implementation of motion control module, explains the theory and categories of interpolation algorithm, proposes optimal sampled data interpolation algorithm and applies it to this system, researches interrupt control and fluorescence signal detection path planning, implements the motion control of fluorescence detection system.
     The fifth chapter describes the implementation of data acquisition and processing module, applies multithreading to this system, processes fluorescence signal include selecting the data segment, digital filter, data smoothing, peak identification, signal normalized, implements monitoring of fluorescence signal, simulates and makes experiment about fluorescence signal monitoring system.
     The last chapter summarizes this paper and prospectes the following research.
引文
[1]邓文星,张映.实时荧光定量PCR技术综述[J].生物技术通报,2007(005):93-95.
    [2]MULLIS K, FALOONA F, SCHARF S, et al. Specific enzymatic amplification of DNA in vitro: the polymerase chain reaction.1986.[J]. Biotechnology (Reading, Mass.),1992,24:17.
    [3]MULLIS K B. Target amplification for DNA analysis by the polymerase chain reaction., 1990[C].
    [4]刘小荣,张笠,王勇平.实时荧光定量PCR技术的理论研究及其医学应用[J].中国组织工程研究与临床康复,2010,14(002):329-332.
    [5]郭琛.荧光定量PCR仪监控系统的设计与实现[D].上海:上海交通大学,2005.
    [6]HEID C A, STEVENS J, LIVAK K J, et al. Real time quantitative PCR.[J]. Genome research, 1996,6(10):986.
    [7]王行,王惠民.实时定量PCR技术的方法学分类[J].临床检验杂志,2007,25(1):71.
    [8]HEID C A, STEVENS J, LIVAK K J, et al. Real time quantitative PCR.[J]. Genome research, 1996,6(10):986.
    [9]WILLARD M F, STEPHEN J W, KENT E V. Quantitative RT-PCR:pitfall and potential [J]. Bio Techniques,1999,26(1):112-125.
    [10]张蓓,沈立松.实时荧光定量PCR的研究进展及其应用[J].国外医学:临床生物化学与检验学分册,2003,24(006):327-329.
    [11]付春华,陈孝平,余龙江.实时荧光定量PCR的应用和进展[J].激光生物学报,2005,14(006):466-471.
    [12]张文超,刘晓光,吴勤勤.基因扩增分析(PCR)仪温控系统的研究与应用[J].华东理工大学学报:自然科学版,2004,30(002):203-206.
    [13]HIGUCHI R, FOCKLER C, DOLLINGER G, et al. Kinetic PCR analysis:real-time monitoring of DNA amplification reactions[J]. Nature Biotechnology,1993,11(9):1026-1030.
    [14]WAHA A, WATZKA M, KOCH A, et al. A rapid and sensitive protocol for competitive reverse transcriptase (cRT) PCR analysis of cellular genes[J]. Brain Pathology,1998,8(1):13-18.
    [15]林玲.定量PCR技术的研究进展[J].国外医学:遗传学分册,1999,22(003):116-120.
    [16]张文超.聚合酶链反应(PCR)技术与基因扩增分析仪器(PCR仪)[J].生命科学仪器,2005,3(003):13-19.
    [17]蔡刚,李闻捷,沈茜.实时定量PCR应用中的问题及优化方案[J].国外医学:临床生物化学与检验学分册,2003,24(006):330-332.
    [18]陈世泽,陈章位,黄靖,等.PCR仪温度控制均匀性研究与仿真分析[J].中国生物医学工程学报,2009,28(4).
    [19]张兰.实时PCR仪中荧光检测模块的研制[D].北京:北京工业大学生物医学工程,2009.
    [20]如何选择合适的定量PCR仪[EB/OL]. [1.4]. http://www.biooo.com/7action-viewnews-itemid-2920.
    [21]邓启辉.基于嵌入式平台的高性能运动控制系统与方法研究[D].武汉理工大学,2005.
    [22]万松峰,黄惟公.基于WinCE&ARM的嵌入式工业监控系统的开发[J].西华大学学报:自然科学版,2005,24(006):26-28.
    [23]张冬泉.Windows CE实用开发技术[M].电子工业出版社,2006.
    [24]周振宇,谷海颖.基于USB总线的实时数据采集系统设计[J].电子技术应用,2002,28(002):17-19.
    [25]倪明辉,周军,杨庚.USB在FPGA控制的高速数据采集系统中的应用[J].计算机测量与控制,2006,14(002):268-271.
    [26]梁颖.基于USB和FPGA技术的高性能数据采集模块的设计与实现[D].四川大学,2005.
    [27]李妮妮.全软件型激光切割机数控系统的研究[D].浙江大学,2010.
    [28]李妮妮,陈章位.基于ARM和CPLD的开放式数控系统设计[J].电子技术应用,2009,35(11).
    [29]MING J. The Features of EZ-USB-FX2and the Implementation of Data Trans fer [J]. Computer Engineering and Applications,2005,11.
    [30]李娜,孟令军.用FT245BM实现FPGA与PC机的数据无缝传输[J].重庆工学院学报:自然科学版,2009,23(005):80-83.
    [31]钱峰. EZ-USB FX2单片机原理,编程及应用[M].北京航空航天大学出版社,2006.
    [32]H. Schildt王军.c++基础教程[M].清华大学出版社,2004.
    [33]吕凤翥.C++语言基础教程[M].清华大学出版社,1999.
    [34]杜四春.C++程序设计[M].中国水利水电出版社,2005.
    [35]孙鑫,余安萍.VC++深入详解[M].电子工业出版社,2006.
    [36]木林森. Visual C++ 6.0使用与开发[M].清华大学出版社,1998.
    [37]VC++6.0[EB/OL]. [12.3]. http://aike.baidu.com/view/149791.htm.
    [38]汪兵,李存斌,陈鹏.EVC高级编程及其应用开发[M].中国水利水电出版社,2005.
    [39]黄新燕.机床数控技术及编程[M].北京理工大学出版社,2006.
    [40]张明亮,解旭辉.开放性数控技术的发展[J].机电工程,1999,16(004):65-67.
    [41]刘雄伟.数控加工理论与编程技术[M].机械工业出版社,2000.
    [42]Luo F Y, ZHOU Y F, YIN J. A universal velocity profile generation approach for high-speed machining of small line segments with look-ahead [J]. The International Journal of Advanced Manufacturing Technology,2007,35(5):505-518.
    [43]严爱珍.机床数控原理与系统[M].机械工业出版社,1999.
    [44]汤阳.基于Windows CE的开放式数控系统关键技术研究[D].合肥工业大学,2003.
    [45]张冬泉. Windows CE实用开发技术[M].电子工业出版社,2006.
    [46]胡朝斌,王治森,董伯麟,等.基于Windows CE的数控系统中断控制[J].中国机械工程,2005,16(009):761-764.
    [47]何宗健. Windows CE嵌入式系统[Z].北京:北京航空航天大学出版社,2006.
    [48]马跃,刘瑞,吴文江.基于Windows CE. NET的数控系统实时性能测试与分析[J].小型微型计算机系统,2007,28(012):2208-2211.
    [49]ZHI-SEN W, GAO E, ZHANG Y, et al. A windows CE-Based open-architectured CNC system [J][J]. Manofacturing Automation,2001,9.
    [50]ZHENGRONG J, ZHISEN W, BOLIN D, et al. Study on the CNC System Based on Windows CE.NET and Its Real-time Interpolation Control [J].Manufacturing Technology Machine Tool,2006,12.
    [51]丁海利,王芳,高成修.旅行商问题的交叉粒子群优化算法[J].数学杂志 2008(001):85-89.
    [52]唐立新.旅行商问题(TSP)的改进遗传算法[J].东北大学学报:自然科学版,1999,20(001):40-42.
    [53]朱献文,李福荣.求解旅行商问题的几种智能算法[J].计算机与数字工程,2010(001):32-35.
    [54]GUTIN G. Traveling Salesman Problem[J].2009.
    [55]APPLEGATE D L. The traveling salesman problem:a computational study[M]. Princeton Univ Pr,2006.
    [56]YUAN Y, YUAN J, Du H, et al. Pareto Ant Colony Algorithm for Building Life Cycle Energy Consumption Optimization[J]. Life System Modeling and Intelligent Computing, 2010:59-65.
    [57]LI N N, DONG Y F, Gu J H, et al. A New Algorithm Based on Immune Algorithm and Hopfield Neural Network for Multimodal Function Optimization,2007[C]. IEEE.
    [58]LEE T F, HSIAO Y C, Wu H Y, et al. Optimization of reactive power compensation and voltage regulation using artificial immune algorithm for radial transmission networks, 2008[C]. IEEE.
    [59]李妮妮,陈章位,陈世泽.基于局部搜索和遗传算法的激光切割路径优化[J]. Computer Engineering and Applications,2010,46(2).
    [60]王岩.一种基于最近邻点法的禁忌搜索算法[J].科技信息,2008,32.
    [61]骆斌,费翔林.多线程技术的研究与应用[J].计算机研究与发展,2000,37(004):407-412.
    [62]周兴铭,徐明.多线程技术的现状与前景展望[J].计算机工程与科学,1999,21(004):1-6.
    [63]马明建,周长城.数据采集与处理技术[M].西安交通大学出版社,1998.
    [64]KIRIANAKI N V, YURISH S Y, SHPAK N O, et al. Data acquisition and signal processing for smart sensors[M]. Wiley Online Library,2002.
    [65]祝常红,彭坚.数据采集与处理技术[Z].北京:电子工业出版社,2008.
    [66]郭俊,李霞,杜星,等.车轮磨耗分析中平滑方法的选取[J].西南交通大学学报, 2010,45(003):373-377.
    [67]KAISER J F, REED W A. Data smoothing using low-pass digital filters[J]. Review of Scientific Instruments,2009,48(11):1447-1457.
    [68]韩丹夫,吴庆标.数值计算方法[M].浙江大学出版社,2006.
    [69]查布拉,卡纳莱,唐玲艳,等.工程数值方法[M].清华大学出版社,2007.
    [70]WELHAM S J, CULLIS B R, KENWARD M G, et al. A comparison of mixed model splines for curve fitting[J]. Australian & New Zealand Journal of Statistics,2007,49(1):1-23.
    [71]LENG Y G, LENG Y S, WANG T Y, et al. Numerical analysis and engineering application of large parameter stochastic resonance[J]. Journal of Sound and Vibration, 2006,292(3-5):788-801.
    [72]MCKINLEY S, LEVINEM. Cubic spline interpolation[J]. Student Projects in Linear Algebra, College of the Redwood, [online] Retrieved November,2006,20.
    [73]CHANG G W, LIU Y J, CHEN C I. Cubic spline interpolation-based method for modeling fluorescent lamp dynamic conductance,2008[C]. IEEE.
    [74]何光渝. Visual c++常用数值算法集[M].科学出版社,2002.
    [75]杨华中,汪蕙.数值计算方法与C语言工程函数库[M].科学出版社,1996.

© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号

地址:北京市海淀区学院路29号 邮编:100083

电话:办公室:(+86 10)66554848;文献借阅、咨询服务、科技查新:66554700