基于类酶活性DNA-Ag/Pt双金属纳米簇的比色法检测血清中尿酸

Colorimetric assay of uric acid in serum based on DNA-Ag/Pt bimetallic nanoclusters with enzyme-like activity

  • 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0.3~75.0 μmol/L,检测限为0.15 μmol/L。选取的健康人血清样本的尿酸含量在203.04~381.40 μmol/L之间,高尿酸患者样本的尿酸含量在420.97~463.27 μmol/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。

     

    Abstract:
    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 H 2O2 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 75 μmol/L, with a detection limit of 0.15 μmol/ L. The concentration of uric acid in selected healthy human serum samples were between 203.04 μmol/L and 381.40 μmol/L, while that of samples from patients with hyperuricemia ranged from 420.97 μmol/L to 463.27 μmol/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.

     

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