摘要
目的:建立一种核酸适配体电化学传感器,用于肿瘤细胞内三磷酸腺苷(ATP)的超灵敏检测。方法:以金电极为基底,通过巯基化学以及DNA自组装技术,以核酸适配体为识别元件,构建一种DNA“三明治”结构,以双重标记的亚甲蓝为电化学探针,通过金纳米颗粒进行信号放大,采用微分脉冲伏安法进行定量。结果:所制备的电化学传感器在10 fmol/L^1 mmol/L范围内,电流信号变化幅度与ATP浓度呈良好的线性关系(R^2=0.9956),最低检测限为29.6 aM(S/N=3)。该电化学传感器对靶标分子特异性强,可准确检测肿瘤细胞中ATP的浓度。结论:本研究所建立的ATP电化学传感器灵敏度高、特异性强且线性范围宽,实现了肿瘤细胞中ATP含量的定量检测,有望为临床相关疾病的分子诊断提供新的技术手段。
Objective:An ultrasensitive ATP aptasensor was presented for detection of ATP level in tumor cells.Methods:A DNA sandwich structure was constructed based on gold electrode,thiol-chemistry,and DNA self-assembly principle,with nucleic acid aptamer as the recognition element.Double-labeled methylene blue(MB)was used as electrochemical tags,and signal was amplified by gold nanoparticles.Quantitative analysis was performed by differential pulse voltammetry(DPV).Results:The prepared electrochemical sensorwas in the range from 10 fmol/L to 1 mmol/L,and the range of current signal change had a good linear relationship with ATP concentration(R2=0.996),with the detection limit of 29.6 aM(S/N=3).The electrochemical sensor had strong specificity on target molecules,which accurately detected the concentration of ATP in the tumor cells.Conclusion:The advantages of the present ATP electrochemical sensor include high sensitivity,strong specificity,as well as broad-range linear response,which realizes the quantitative detection of ATP content in tumor cells,thus holding a potential to be a new technical mean for molecular diagnosis of clinically-related diseases.
作者
李书婷
杨泽奇
陈燕宏
陈丽敏
李新春
Li Shuting;Yang Zeqi;Chen Yanhong;Chen Limin;Li Xinchun(School of Pharmacy,Guangxi Medical University,Nanning 530021,China)
出处
《广西医科大学学报》
CAS
2020年第12期2276-2281,共6页
Journal of Guangxi Medical University
基金
国家自然科学基金资助项目(No.82060647)
广西研究生教育创新项目(No.YCSW2020126)。
