PDF全文下载：2011050478
邹元明， 冷翠翠， 万钧*
（青岛科技大学 化学与分子工程学院，山东 青岛 266042）摘要: 以聚苯胺纳米管、壳聚糖、室温离子液体修饰的丝网印刷电极为基底电极，以辣根过氧化物酶（HRP）催化的H2O2氧化邻苯二胺（OPD）的反应作为信号放大手段，构建了一个酶放大丝网印刷DNA传感器。考察并确定了在修饰丝网印刷电极上，上述反应的最优条件及进行电化学测定的最优条件：HRP催化OPD与H2O2反应的最佳缓冲液为pH 5.0的醋酸-醋酸钠缓冲液；最佳检测支持电解质为pH 7.2的PBS缓冲液。HRP催化OPD与双氧水反应的最佳反应条件：使用16 mmol·L-1双氧水和30 mmol·L-1 OPD反应15 min。在该传感器上，在最优条件下，DNA测定的线性范围为2×10-14 mol·L-1到8×10-13 mol·L-1，回归方程为Δip=0.308 6+0.016 0 cT。通过3σ规则计算出的DNA检测限为8×10-15 mol·L-1。该传感器在DNA检测方面具有良好的选择性。
关键词: 修饰丝网印刷电极； 辣根过氧化物酶； 邻苯二胺； DNA传感器
中图分类号: O 657.1；Q 523文献标志码： AA Screen-printed Electrode DNA Sensor Based on Horseradish
Peroxidase-catalysed AmplificationZOU Yuan-ming, LENG Cui-cui, WAN Jun
（College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao 266042, China)Abstract: In this study, a DNA sensor was constructed based on the screen-printed electrode that was modified with polyaniline nanotubes, chitosan and room-temperature ionic liquids, and by using the horseradish peroxidase catalyzed oxidation of H2O2 to o-phenenyldiamine (OPD) as the signal amplification approach. The optimal conditions for the above-mentioned reaction and for the electrochemical determination on the modified screen-printed electrode were established as follows: the optimal buffer for the HRP-catalyzed oxidation of OPD by H2O2 was HOAc-NaOAc at pH 5.0; the optimal buffer for the electrochemical determination was PBS at pH 7.2; the best working condition for HRP-catalyzed oxidation was at a reaction time of 15 min with 16 mmol·L-1 H2O2 and 30 mmol·L-1 OPD. Under these optimal conditions, DNA can be determined in a linear range from 2×10-14 mol·L-1 to 8×10-13 mol·L-1, with a regression equation Δip=0.308 6+0.016 0cT. The detected limit was found to be 8×10-15 mol·L-1 according to 3σ rule. Good selectivity against mismatched DNAs was verified.
Key words: modified screen-printed electrode; horseradish peroxidase; o-phenenyldiamine; DNA sensor
收稿日期:2011-03-07
作者简介: 邹元明（1985—），男，硕士研究生.*通信联系人.
