Citation: HAI Hong, HUANG Wen-Gang, LIANG Shun-Chao, LI Jian-Ping. Ultrasensitive Electrochemiluminescence Biosensor for DNA Determination Based on Nicking Endonuclease Assisted Signal Amplification[J]. Chinese Journal of Analytical Chemistry, ;2016, 44(5): 779-786. doi: 10.11895/j.issn.0253-3820.150742 shu

Ultrasensitive Electrochemiluminescence Biosensor for DNA Determination Based on Nicking Endonuclease Assisted Signal Amplification

Guangxi Key Laboratory of Electrochemical and Magnetochemical Function Materials, College of Chemistry and Bioengineering, Guilin University of Technology, Guilin 541004, China

Received Date: 19 September 2015
Revised Date: 29 December 2015

Fund Project: This work was supported by the National Natural Science Foundation of China (Nos.21165007, 21375031)

A novel electrochemiluminescence (ECL) DNA biosensor based on nicking endonuclease and the efficient ECL property of quantum dots(QDs) assisted signal amplification was developed. The capture probe DNA(c-DNA) was self-assembled on gold electrode via AuS bond, and then hybridized with target DNA (t-DNA) to form double-stranded DNA. Then nicking endonuclease Nt.BstNBI recognized a cutting site (5'-GAGTC-3') in the double chain and cleaved c-DNA strand at 4 bases away from the 3' end of its recognition site, thus releasing the t-DNA and achieving a recycle of t-DNA in the next hybridization and signal amplification. Finally, carboxyl groups on the surface of CdTe QDs were activated with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and N-hydroxysuccinimide (NHS), and then reacted with amino groups at the terminal of residual c-DNA on the electrode surface. So the QDs were fabricated and the t-DNA concentration could be determined by measuring the ECL signal of the CdTe QDs. The experimental conditions were optimized and 1 μmol/L c-DNA, 60 min of hybridization time, 0.5 U/μL Nt.BstNBI and 4 h of endonuclease reaction time were chosen. The experimental results showed that under optimal conditions, t-DNA could be specifically assayed with a linear relationship between the ECL signal intensity and the logarithm of t-DNA concentration in the range of 2.0×10^-13-2.0×10^-11 mol/L, with a limit of detection of 7.3×10^-14 mol/L. The biosensor was successfully applied to determine t-DNA concentration in human blood sample with recoveries of 96.4%-108.0%.
- DNA biosensor,
- Nicking endonuclease,
- Quantum dots,
- Electrochemiluminescence,
- Signal amplification
1. [1]
2. [2]
3. [3]
4. [4]
5. [5]
6. [6]
7. [7]
8. [8]
9. [9]
10. [10]
11. [11]
12. [12]
13. [13]
14. [14]
15. [15]
16. [16]
17. [17]
18. [18]
19. [19]
20. [20]
21. [21]
22. [22]
23. [23]
24. [24]
25. [25]
26. [26]
27. [27]
28. [28]
29. [29]
30. [30]
31. [31]
32. [32]
33. [33]
34. [34]
35. [35]
36. [36]
37. [37]
38. [38]
1. [1]
  Miaomiao He , Zhiqing Ge , Qiang Zhou , Jiaqing He , Hong Gong , Lingling Li , Pingping Zhu , Wei Shao . Exploring the Fascinating Realm of Quantum Dots. University Chemistry, 2024, 39(6): 231-237. doi: 10.3866/PKU.DXHX202310040
2. [2]
  Yu SU , Xinlian FAN , Yao YIN , Lin WANG . From synthesis to application: Development and prospects of InP quantum dots. Chinese Journal of Inorganic Chemistry, 2024, 40(11): 2105-2123. doi: 10.11862/CJIC.20240126
3. [3]
  Zeyu XU , Anlei DANG , Bihua DENG , Xiaoxin ZUO , Yu LU , Ping YANG , Wenzhu YIN . Evaluation of the efficacy of graphene oxide quantum dots as an ovalbumin delivery platform and adjuvant for immune enhancement. Chinese Journal of Inorganic Chemistry, 2024, 40(6): 1065-1078. doi: 10.11862/CJIC.20240099
4. [4]
  Meiqing Yang , Lu Wang , Haozi Lu , Yaocheng Yang , Song Liu . Recent Advances of Functional Nanomaterials for Screen-Printed Photoelectrochemical Biosensors. Acta Physico-Chimica Sinica, 2025, 41(2): 100018-. doi: 10.3866/PKU.WHXB202310046
5. [5]
  Jiarong Feng , Yejie Duan , Chu Chu , Dezhen Xie , Qiu'e Cao , Peng Liu . Preparation and Application of a Streptomycin Molecularly Imprinted Electrochemical Sensor: A Suggested Comprehensive Analytical Chemical Experiment. University Chemistry, 2024, 39(8): 295-305. doi: 10.3866/PKU.DXHX202401016
6. [6]
  Qiaoqiao BAI , Anqi ZHOU , Xiaowei LI , Tang LIU , Song LIU . Construction of pressure-temperature dual-functional flexible sensors and applications in biomedicine. Chinese Journal of Inorganic Chemistry, 2024, 40(12): 2259-2274. doi: 10.11862/CJIC.20240128
7. [7]
  Minna Ma , Yujin Ouyang , Yuan Wu , Mingwei Yuan , Lijuan Yang . Green Synthesis of Medical Chemiluminescence Reagents by Photocatalytic Oxidation. University Chemistry, 2024, 39(5): 134-143. doi: 10.3866/PKU.DXHX202310093
8. [8]
  Jianjun Liu , Xue Yang , Chi Zhang , Xueyu Zhao , Zhiwei Zhang , Yongmei Chen , Qinghong Xu , Shao Jin . Preparation and Fluorescence Characterization of CdTe Semiconductor Quantum Dots. University Chemistry, 2024, 39(7): 307-315. doi: 10.3866/PKU.DXHX202311031
9. [9]
  Li'na ZHONG , Jingling CHEN , Qinghua ZHAO . Synthesis of multi-responsive carbon quantum dots from green carbon sources for detection of iron ions and L-ascorbic acid. Chinese Journal of Inorganic Chemistry, 2025, 41(4): 709-718. doi: 10.11862/CJIC.20240280
10. [10]
  Borong Yu , Huijiao Zhang , Xinyu Zhang , Xiaoying Li , Shuming Chen , Zhangang Han . The Blue Elf in the Dark: Gradient Science Popularization Experiments on Chemiluminescence. University Chemistry, 2024, 39(9): 295-303. doi: 10.12461/PKU.DXHX202403107
11. [11]
  Xingchao Zhao , Xiaoming Li , Ming Liu , Zijin Zhao , Kaixuan Yang , Pengtian Liu , Haolan Zhang , Jintai Li , Xiaoling Ma , Qi Yao , Yanming Sun , Fujun Zhang . 倍增型全聚合物光电探测器及其在光电容积描记传感器上的应用. Acta Physico-Chimica Sinica, 2025, 41(1): 2311021-. doi: 10.3866/PKU.WHXB202311021
12. [12]
  Wenkai Chen , Yunjia Shen , Xiangmeng Kong , Yanli Zeng . Quantum Chemistry Calculation of Key Physical Quantity in Circularly Polarized Luminescence: Introducing an Exploratory Computational Chemistry Experiment. University Chemistry, 2025, 40(3): 83-91. doi: 10.12461/PKU.DXHX202405018
13. [13]
  Jing SU , Bingrong LI , Yiyan BAI , Wenjuan JI , Haiying YANG , Zhefeng Fan . Highly sensitive electrochemical dopamine sensor based on a highly stable In-based metal-organic framework with amino-enriched pores. Chinese Journal of Inorganic Chemistry, 2024, 40(7): 1337-1346. doi: 10.11862/CJIC.20230414
14. [14]
  Zhenjun Mao , Haorui Gu , Haiyan Che , Xufeng Lin . Exploration on Experiment Teaching of UHPLC-IC Based on Valve Switching Method. University Chemistry, 2024, 39(4): 81-86. doi: 10.3866/PKU.DXHX202311013
15. [15]
  Jinghan Xu , Yang Wang , Donghui Wei . Drawing Cross-Sectional Contour Maps of π Molecular Orbitals. University Chemistry, 2025, 40(3): 23-29. doi: 10.12461/PKU.DXHX202403023
16. [16]
  Hongyan Feng , Weiwei Li . Reflections on the Safety of Chemical Science Popularization Activities. University Chemistry, 2024, 39(9): 379-384. doi: 10.12461/PKU.DXHX202404087
17. [17]
  Yanan Jiang , Yuchen Ma . Brief Discussion on the Electronic Exchange Interaction in Quantum Chemistry Computations. University Chemistry, 2025, 40(3): 10-15. doi: 10.12461/PKU.DXHX202402058
18. [18]
  Yaqin Zheng , Lian Zhuo , Meng Li , Chunying Rong . Enhancing Understanding of the Electronic Effect of Substituents on Benzene Rings Using Quantum Chemistry Calculations. University Chemistry, 2025, 40(3): 193-198. doi: 10.12461/PKU.DXHX202406119
19. [19]
  Jiabo Huang , Quanxin Li , Zhongyan Cao , Li Dang , Shaofei Ni . Elucidating the Mechanism of Beckmann Rearrangement Reaction Using Quantum Chemical Calculations. University Chemistry, 2025, 40(3): 153-159. doi: 10.12461/PKU.DXHX202405172
20. [20]
  Dongju Zhang , Rongxiu Zhu . Construction of Ideological and Political Education in Quantum Chemistry Course: Several Teaching Cases to Reveal the Universal Connection of Things. University Chemistry, 2024, 39(7): 272-277. doi: 10.3866/PKU.DXHX202311032

Get Citation

PDF

XML

Metrics

PDF Downloads(0)
Abstract views(653)
HTML views(110)

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142