摘要
目的:利用类酶活性DNA-Ag/Pt双金属纳米簇(DNA-Ag/Pt NCs)建立血清中尿酸含量检测的比色传感新方法。方法:首先,以单链DNA为模板制备具有高类过氧化物酶活性的DNA-Ag/Pt NCs,并通过透射电子显微镜、动态光散射、Zeta电位、X光电子能谱、X射线衍射仪、顺磁共振波谱仪和傅里叶红外光谱对其进行表征。利用DNA-Ag/Pt NCs可催化尿酸与尿酸酶的反应产物H2O2与3,3',5,5'-四甲基联苯胺发生显色反应的特点,根据溶液颜色变化及紫外—可见分光光度计测定结果定量检测尿酸含量。将该方法应用于人血清样本中的尿酸检测和加标回收实验。结果:成功制备得到具有高类过氧化物酶活性的DNA-Ag/Pt NCs。所建立方法用于选择性尿酸检测的线性范围为0.3~75.0μmol/L,检测限为0.15μmol/L。选取的健康人血清样本的尿酸含量在203.04~381.40μmol/L之间,高尿酸患者样本的尿酸含量在420.97~463.27μmol/L之间,加标回收率为95.0%~109.8%。结论:基于DNA-Ag/Pt NCs建立的尿酸比色测定法具有较高的特异性与灵敏度,并成功实现了血清样品中尿酸的检测。
Objective:To establish a novel colorimetric sensor for uric acid in serum via DNA-Ag/Pt bimetallic nanoclusters(DNA-Ag/Pt NCs)with enzyme-like activity.Methods:Firstly,DNA-Ag/Pt NCs with high peroxidase-like activity were synthesized using single-stranded DNA as the template.The DNA-Ag/Pt NCs were characterized by transmission electron microscopy,dynamic light scattering,Zeta potential,X-ray photoelectron spectroscopy,X-ray diffractometer,electron resonance spectroscopy and Fourier transform infrared spectroscopy.Then,DNA-Ag/Pt NCs catalyzed the color reaction between H2O2 and 3,3’,5,5’-tetramethylbenzidine,in which H2O2 was generated by the reaction of uricase and UA.The concentration of uric acid was quantitatively determined based on the color changes and the measurement results by UV-vis spectrometer.Furthermore,this method was applied to the detection of uric acid concentration and the standard addition recovery experiments in human serum samples.Results:DNA-Ag/Pt NCs with high peroxidase-like activity were successfully prepared.This method showed a linear detection range for uric acid from 0.3 to 75μmol/L,with a detection limit of 0.15μmol/L.The concentration of uric acid in selected healthy human serum samples were between 203.04μmol/L and 381.40μmol/L,while that of samples from patients with hyperuricemia ranged from 420.97μmol/L to 463.27μmol/L.The standard addition recovery rate was in the range from 95.0%to 109.8%.Conclusion:The colorimetric assay based on DNA-Ag/Pt NCs for uric acid exhibits high specificity and sensitivity,and successfully achieves the detection of uric acid in serum samples.
作者
洪婉蓉
潘向和
覃英凤
刘金文
HONG Wanrong;PAN Xianghe;QIN Yingfeng;LIU Jinwen(School of Basic Medical Sciences,Key Laboratory of Biological Molecular Medicine Research,Education Department of Guangxi Zhuang Autonomous Region,Guangxi Medical University,Nanning 530021,China;Key Laboratory of Longevity and Aging-related Diseases Ministry of Education,Guangxi Medical University,Nanning 530021,China)
出处
《广西医科大学学报》
2025年第3期377-384,共8页
Journal of Guangxi Medical University
基金
国家自然科学基金资助项目(No.22266007)
广西自然科学基金资助项目(No.2024GXNSFAA010208)
长寿与老年相关疾病教育部重点实验室资助项目(No.KLLAD202302)。
