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Citation: LI Chen-Wei, LIN Sheng-Hao, DU Zai-Hui, SUN Chun-Yan, XU Wen-Tao. Establishment and Application of Lead Ion Functional Nucleic Acid Colorimetric Biosensor[J]. Chinese Journal of Analytical Chemistry, ;2019, 47(9): 1427-1432. doi: 10.19756/j.issn.0253-3820.191217 shu

Establishment and Application of Lead Ion Functional Nucleic Acid Colorimetric Biosensor

1.
College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China;
2.
College of Food Science and Engineering, Jilin University, Changchun 130062, China

Received Date: 19 April 2019
Revised Date: 5 July 2019

Fund Project: This work was supported by the National Natural Science Foundation of China (No.31871875).

A method for visual detection of Pb²⁺ in water based on lead ion (Pb²⁺) functional nucleic acid colorimetric biosensor was developed. The specific recognition and cleavage of Pb²⁺ by GR-5 deoxyribozyme was carried out, and the cleavage products of GR-5 deoxyribozyme were combined with the color templates to convert into a G-quadruplex by clever nucleic acid design, wherein the color templates mainly included a complementary region to the cleavage product nucleic acid sequences, a guanine-rich G quadruplet forming region, and a spacer arm. In the presence of Pb²⁺, the substrate chain of the GR-5 deoxyribozyme was recognized to be cleaved and released, and the cleavage product was converted into a G-quadruplex by binding to a chromogenic template, since the G-quadruplex was under specific conditions. The peroxidase-like activity catalyzed the color development of H₂O₂ and 3,3',5,5'-tetramethylbenzidine (TMB), thus completing the construction of the Pb²⁺ functional nucleic acid colorimetric biosensor. Through a series of conditions optimization, it was found that the optimal ratio of substrate chain and enzyme chain of GR-5 deoxyribozyme was 1:1, the cleavage time was 3 min, the color template concentration was 5 μmol/L, and the optimal TMB concentration was 50 μL. Under the optimal conditions, the detection range of Pb²⁺ was 25 nmol/L-2.5 μmol/L, the linear relationship was y=0.039x+0.3486 (R=0.9904), and the lowest detection concentration was 10.1 nmol/L (3σ). The recoveries were 98.2%-115.5%. The proposed method had a good application prospect due to its high sensitivity, strong specificity and simple operation.
- Lead ion (Ⅱ),
- Functional nucleic acid,
- Visualization,
- G-quadruplex,
- Biosensor
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