Citation: ZHANG He, WANG Qing, FU Xin, ZHAO Zhi-Liang. Fabrication of “Turn-On” Sensor for Sensitive Detection of Hg²⁺ Based on Rolling Circle Amplification and Tandem G-quadruplex-hemin DNAzymes[J]. Chinese Journal of Analytical Chemistry, ;2018, 46(10): 1644-1651. doi: 10.11895/j.issn.0253-3820.181346 shu

Fabrication of “Turn-On” Sensor for Sensitive Detection of Hg²⁺ Based on Rolling Circle Amplification and Tandem G-quadruplex-hemin DNAzymes

School of Chemistry and Chemical Engineering, Hunan Institute of Engineering, Xiangtan 411104, China

Received Date: 24 May 2018
Revised Date: 11 July 2018

Fund Project: This work was supported by the National Natural Science Foundation of China (No.21005067) and the Scientific Research Fund of Hunan Provincial Education Department (Nos.17A044, 16B060).

Based on the tandem G-quadruplex-hemin DNAzymes for catalytic signal amplification and T-Hg²⁺-T structure for specific recognition, a highly sensitive "Turn-on" biosensor for detection of mercury ion (Hg²⁺) was developed on the polystyrene microbeads to achieve the detection of Hg²⁺ in urine samples. Through the specific binding of streptavidin and biotin, the T-rich biotinylated Hg²⁺ capture probe was immobilized on the surface of the microspheres. When Hg²⁺ was present in reaction system, the T-rich Hg²⁺ capture probe could capture the cyclic DNA containing G-quadruplex complementary fragments to the surface of beads by forming T-Hg²⁺-T structure. Through rolling circle amplification, a large number of tandem G-quadruplex forming sequences were amplified. When hemin was inserted into the G-quadruplex, a G-quadruplex-hemin DNAzyme with enhanced catalytic activity was formed, which could catalyze the oxidation of ABTS to ABTS^·+ under the condition of H₂O₂. Finally, the absorbance difference at 420 nm was detected by UV-vis spectrophotometer. Under the optimal experimental conditions, the linear range for detection of Hg²⁺ was 0.4 pmol/L-100 pmol/L with the detection limit of 0.3 pmol/L, and the regression equation was ΔA_{420 nm}=0.1+0.0019C_Hg²⁺ (pmol/L). This Hg²⁺ sensing system had excellent selectivity for Hg²⁺ against other possible competing ions. The fabricated sensor was applied to detection of Hg²⁺ in urine samples, the relative standard deviation and the recovery were 1.4%-2.6% and 94.0%-106.0%, respectively. The sensor had good selectivity, sensitivity, anti-interference ability and reliable detection capability for complex practical samples.
- G-quadruplex-hemin DNAzyme,
- Rolling circle amplification,
- Mercury ion (II),
- Urine analysis,
- Colorimetric analysis
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通讯作者: 陈斌, bchen63@163.com

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Fabrication of “Turn-On” Sensor for Sensitive Detection of Hg2+ Based on Rolling Circle Amplification and Tandem G-quadruplex-hemin DNAzymes

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通讯作者: 陈斌, bchen63@163.com

Fabrication of “Turn-On” Sensor for Sensitive Detection of Hg²⁺ Based on Rolling Circle Amplification and Tandem G-quadruplex-hemin DNAzymes