基于均相电化学的新型生物传感技术研究
发布时间：2018-12-12 15:39
【摘要】：近年来,食品卫生与安全及毒素的检测成为备受关注的热门话题,目前针对食品真菌毒素检测,迫切需要高灵敏的检测技术,从而保障消费安全。传统的生物传感技术一般需要繁琐的标记过程,且灵敏度较差,本论文针对这些问题,结合非标记均相传感器的优点以及能与目标靶分子特异性结合的核酸适配体的优势发展出了两种灵敏度高、选择性好的新方法用于检测食品中毒素。主要内容如下:(1)提出了一种非标记均相电化学方法用于来检测黄曲霉毒素B1。该方法旨在以ITO电极表面长链DNA和短链DNA扩散性质的不同和核酸外切酶的性质相结合用于信号转换系统。当黄曲霉毒素B1存在时,其与核酸适配体反应释放的单链被外切酶切割扩散到电极表面,能观察到明显的电化学信号。(2)利用二茂铁和亚甲基蓝为信号指示剂的“信号开启”和“信号关闭”体系,发展了一种新型的在均相溶液中反应的可检测多种毒素的电化学适配体生物传感器。该方法设计了两种与毒素有关的单链DNA通过金硫键自组装固定到金电极表面,当在均相溶液中加入两种毒素时,这种双链结构就会解离,并通过形成毒素与适配体复合物释放出信号指示剂标记的DNA,来产生电化学信号。该方法的选择性好,灵敏度高,成本低。其检测OTA和AFB1的检测下限分别为:0.04ng/mL、0.01ng/mL。
[Abstract]:In recent years, the detection of food hygiene and safety and toxins has become a hot topic of much concern. At present, high sensitive detection technology is urgently needed for the detection of food fungal toxins, so as to ensure the safety of consumption. Traditional biosensor technology usually requires tedious labeling process, and its sensitivity is poor. In this paper, we aim at these problems. Combining the advantages of heterogeneous sensors and the advantages of aptamers that can specifically bind to target molecules, two new sensitive and selective methods have been developed for the detection of toxins in food. The main contents are as follows: (1) A heterogeneous electrochemical method for the detection of aflatoxin B _ 1 was proposed. The purpose of this method is to combine the diffusion properties of long chain DNA and short chain DNA on the surface of ITO electrode and the properties of nucleic acid exonuclease to signal conversion system. When aflatoxin B1 exists, the single strand released by its reaction with aptamer of nucleic acid is cut by exonuclease and diffused to the electrode surface. Obvious electrochemical signals can be observed. (2) "signal on" and "signal off" systems using ferrocene and methylene blue as signal indicators, A novel electrochemical aptamer biosensor for the detection of multiple toxins in homogeneous solution was developed. In this method, two single-stranded DNA associated with toxins were designed to be self-assembled by gold-sulfur bond to the surface of the gold electrode. When two toxins were added into the homogeneous solution, the double-stranded structure would be dissociated. The electrochemical signal was generated by the formation of the toxin and aptamer complex and the release of signal indicator labeled DNA,. The method has good selectivity, high sensitivity and low cost. The detection limits of OTA and AFB1 were 0.04ng / mL and 0.01ng / mL, respectively.
【学位授予单位】：温州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：TP212.3;TS207.5
本文编号：2374835
[Abstract]:In recent years, the detection of food hygiene and safety and toxins has become a hot topic of much concern. At present, high sensitive detection technology is urgently needed for the detection of food fungal toxins, so as to ensure the safety of consumption. Traditional biosensor technology usually requires tedious labeling process, and its sensitivity is poor. In this paper, we aim at these problems. Combining the advantages of heterogeneous sensors and the advantages of aptamers that can specifically bind to target molecules, two new sensitive and selective methods have been developed for the detection of toxins in food. The main contents are as follows: (1) A heterogeneous electrochemical method for the detection of aflatoxin B _ 1 was proposed. The purpose of this method is to combine the diffusion properties of long chain DNA and short chain DNA on the surface of ITO electrode and the properties of nucleic acid exonuclease to signal conversion system. When aflatoxin B1 exists, the single strand released by its reaction with aptamer of nucleic acid is cut by exonuclease and diffused to the electrode surface. Obvious electrochemical signals can be observed. (2) "signal on" and "signal off" systems using ferrocene and methylene blue as signal indicators, A novel electrochemical aptamer biosensor for the detection of multiple toxins in homogeneous solution was developed. In this method, two single-stranded DNA associated with toxins were designed to be self-assembled by gold-sulfur bond to the surface of the gold electrode. When two toxins were added into the homogeneous solution, the double-stranded structure would be dissociated. The electrochemical signal was generated by the formation of the toxin and aptamer complex and the release of signal indicator labeled DNA,. The method has good selectivity, high sensitivity and low cost. The detection limits of OTA and AFB1 were 0.04ng / mL and 0.01ng / mL, respectively.
【学位授予单位】：温州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：TP212.3;TS207.5
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