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基于细菌DNA酶滚环扩增检测大肠杆菌

发表时间:2019-03-22  浏览量:727  下载量:59
全部作者: 江西,郑小芳,王娅娅,宦双燕
作者单位: 湖南大学化学化工学院
摘 要: 为实现对大肠杆菌的高效、灵敏检测,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的构型改变,激活催化活性,还可作为锁式探针合成环形模板进行滚环扩增反应。并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103~108 cfu/mL浓度范围的大肠杆菌K12菌株进行有效检测。
关 键 词: 分析化学;大肠杆菌;DNA酶;滚环扩增;SYBR Green I
Title: Detection of E. coli based on bacterial DNAzyme rolling circle amplification
Author: JIANG Xi, ZHENG Xiaofang, WANG Yaya, HUAN Shuangyan
Organization: College of Chemistry and Chemical Engineering, Hunan University
Abstract: To realize an efficient and sensitive detection on E. coli, in this paper, the bacterial DNAzyme and the rolling circle amplification technology are combined to develop a novel method of biological analysis on E. coli, based on the DNAzyme’s high specificity recognition as an aptamer and the high catalytic as a biological enzyme. In this strategy, DNAzyme not only recognizes the structural changes in target protein 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 show that the method of detecting E. coli based on DNAzyme self-cleavage and rolling circle amplification technology, can effectively detect E. coli K12 in the concentration range of 103-108 cfu/mL.
Key words: analytical chemistry; E. coli; DNAzyme; rolling circle amplification; SYBR Green I
发表期数: 2019年2月第1期
引用格式: 江西,郑小芳,王娅娅,等. 基于细菌DNA酶滚环扩增检测大肠杆菌[J]. 中国科技论文在线精品论文,2019,12(1):99-107.
 
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