基于DNA循环放大肿瘤标志物传感分析研究.doc

硕 士 学 位 论 文 MASTER DEGREE THESIS 基于DNA循环放大的肿瘤标志物传感分析研究 摘要 本论文的主要内容是将电化学分析技术、分析技术、化学发光分析技术、纳米技术DNA杂交技术相结合，研制出高灵敏度、高选择性的新型DNA生物传感器，对部分肿瘤标志物进行选择性的识别和测定，为肿瘤的临床早期诊断提供基础性研究资料。本文着重了三种类型的DNA生物传感器的研制性质研究： 1. 研究了一种新颖灵敏的将用于检测DNA的生物传感器。工作原理如下：的浓度可通过确定，目标DNA的浓度与的浓度成正比，从而实现目标DNA的检测。在实验的最佳条件下，得到了目标DNA的线性范围为.0 × 10–16 ~1.0 × 10–15 mol/L，检测限为×10?16 M。2.在结合磁珠和酶循环放大的前期工作基础上，我们又研究了一种结合聚合酶循环放大、熵驱动过程和适体识别手段的ATP生物传感器。它用了比色检测手段对Ramos癌细胞中的ATP含量进行了检测。传感器工作原理如下：H2O2显色，形成的OPDA-H2O2-Avidin-HRP体系Avidin-HRP含量，即，OPDA-H2O2显色程度有关，并在一定范围内成一定比例关系，所以可用于检测ATP含量。在最佳实验条件下，对进行了测定 × 10-10 M。并且在此基础上，实现了癌细胞中ATP含量的检测。 3. 在以上两种传感器的研究基础上，我们结合必威体育精装版的文献报道探索了，与前两种不同的是这新的的研究目的不再局限于DNA的识别和检测，它的主要用途检测。链酶亲和素包被板在最佳实验条件下，对进行了测定关键词：；；细胞 ABSTRACT The main idea of the this paper was to explore a novel and sensitive biosensor for the determination of tumor markers based on electrochemical, colorimetric, chemiluminescence, nanotechnology, DNA hybridization，DNA cycling amplification. The biosensor we developed can selectively recognize and detect some kinds of biomarker for tumor, which provides basic research information to early diagnosis of cancer. In this paper, three types of DNA biosensors are summarized as follow: 1. A novel and sensitive electrochemical biosensors based on DNA material and the enzyme cycle amplification for the detection of target DNA has been developed. The Work principals are as follows: Firstly, the assembly process of the DNA sensor. the DNA skeleton is attached to the beads in order to expand the DNA probes on the modified quantum dots. Secondly, Cycle amplification based on restriction endonuclease. The detection sensitivity of the DNA sequence was further improved by the cycle amplification process of the DNA biosensor built with restriction endonuclease. Finally, the electrochemical detection process. We added PbS quantum dots into the reaction. Followed by the nitric acid treation, the concentration of target DNA is proportional to the concentration of Pb2+ within a certain range, we can dete