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化学学报
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化学学报 ›› 2011, Vol. 69 ›› Issue (20): 2493-2498. 上一篇    下一篇

研究论文

非标记纳米银探针催化共振散射光谱检测痕量ATP

欧阳辉祥，刘庆业，梁爱惠，蒋治良*   

  1. (广西师范大学环境与资源学院 桂林 541004)
  • 投稿日期:2011-04-04 修回日期:2011-05-30 发布日期:2011-06-20
  • 通讯作者: 蒋治良 E-mail:zljiang@mailbox.gxnu.edu.cn

Nanocatalytic Resonance Scattering Spectral Detection of Trace ATP Using Labeled-free Nanosilver as Probe

OUYang-Hui-Xiang , LIU Qing-Ye, LIANG Ai-Hui, JIANG Zhi-Liang   

  1. (School of Environment and Resource, Guangxi Normal University, Guilin 541004)
  • Received:2011-04-04 Revised:2011-05-30 Published:2011-06-20

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1431

被引次数

13

在pH 7.8 Tris-HCl缓冲液中, 三磷酸腺苷(adenosine triphosphate, ATP)的核酸适体(Apt 1)与其互补链(Apt 2)结合生成双链DNA (double-strand DNA, dsDNA). 此dsDNA不能稳定纳米银(AgNP), NaCl可致AgNP聚集, 在500 nm波长处产生一个较强的共振散射峰. 加入ATP后, ATP与dsDNA中的Apt 1结合形成较稳定的发夹结构结合物并释放出可稳定AgNP的Apt 2. 随着ATP浓度(16.5～1650 nmol/L)增加, 生成的Apt 2增加, 被Apt 2稳定的AgNP即AgNP-Apt 2结合物增加, 聚集的AgNP减少, 500 nm处的共振散射值线性减小. 该适配体反应中的AgNP-Apt 2对葡萄糖-铜(II)微粒反应具有较强的催化作用, 其产物氧化亚铜微粒在610 nm处有一较强共振散射峰. 随着ATP浓度增大, 反应液中AgNP-Apt 2增多, 催化作用增强, 610 nm处的共振散射峰增强. ATP浓度在4.95～165 nmol/L范围内与共振散射增大值ΔI610 nm呈线性关系, 检出限为1.8 nmol/L ATP. 据此建立了灵敏度高、选择性好、简便快速检测ATP的共振散射光谱新方法.

关键词: ATP, 适配体, 纳米银催化, 共振散射光谱法

n pH 7.8 Tris-HCl buffer solution, the aptamer (Apt 1) combined with its complementary aptamer (Apt 2) to form double-strand DNA (dsDNA) that can not protect nanosilver (AgNP). Under the action of NaCl, the AgNP particles were aggregated to large particles that exhibited a resonance scattering peak at 500 nm. Upon addition of adenosine triphosphate (ATP), it reacted with the Apt 1 of dsDNA to produce stable ATP-Apt 1 conjugate, and the released Apt 2 combined with AgNP to form stable AgNP-Apt 2 conjugate. When ATP concentration increased from 16.5 to 1650 nmol/L, the stable AgNP-Apt 2 increased, and the aggregated AgNP decreased. Thus, the RS intensity at 500 nm decreased linearly. Results showed that the AgNP-Apt 2 exhibited strong catalytic effect on the Cu2O particle reaction between glucose and Cu(II), and the product of Cu2O particles appeared a RS peak at 610 nm. When ATP concentration increased, the AgNP-Apt 2 increased, the RS intensity at 610 nm enhanced linearly. The enhanced RS intensity was linear to ATP concentration in the range of 4.95～165 nmol/L, with a detection limit of 1.8 nmol/L ATP. Thus, a highly sensitive, selective and simple RS assay was proposed for ATP.

Key words: ATP, aptamer, nanosilver catalysis, resonance scattering spectral assay