基于细菌DNA酶滚环扩增检测大肠杆菌
首发时间:2018-10-19
摘要:为实现对大肠杆菌的高效、灵敏检测,本文基于细菌DNA酶,利用其作为核酸适配体的高特异性识别功能和作为生物酶的高催化性能,结合滚环扩增技术,用于大肠杆菌的生物分析新方法研究。在此方案中,DNA酶不仅能够识别目标蛋白发生构型改变,激活催化活性,还将作为锁式探针合成环形模板进行滚环扩增反应。通过实验对滚环扩增的时长、DNA链的反应比例等条件进行优化。结果表明,基于DNA酶自剪切和滚环扩增技术检测大肠杆菌的方法能够对10^3?10^8 cfu / mL 浓度范围的大肠杆菌进行有效检测。
关键词: 大肠杆菌 DNA酶 滚环扩增 SYBR Green I
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Detection of E.coli based on DNAzyme Rolling Circle Amplification
Abstract:To realize a sensitive and efficent detection of E.coli, in this paper, the bacterial DNAzyme and the rolling circle amplification technology are combined to develop a novel analysis method, based on the DNAzyme\'s high recongnition as an aptamer and the high catalytic as a enzyme. In this strategy, DNAzyme not only recognizes the target protein generate configuration changes and then activates catalytic activity, but also serves as a lock probe to synthesize a circular template for rolling circle amplification. The reaction time of the rolling circle amplification, the ratio of DNA chains were optimized in the experiment. The results showed that the method of detecting Escherichia coli based on DNAzyme self-cleavage and rolling circle amplification technology can effectively detect E. coli in the concentration range of 10^3 ~ 10^8 cfu /mL.
Keywords: E.coli DNAzyme;Rolling Circle Amplification SYBR Green I
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