Citation: ZHANG Meng, HAI Hong, ZHOU Fen-Yue, ZHONG Jing-Cai, LI Jian-Ping. Electrochemical Luminescent DNA Sensor Based on Polymerase-assisted Signal Amplification[J]. Chinese Journal of Analytical Chemistry, ;2018, 46(2): 203-210. doi: 10.11895/j.issn.0253-3820.171020 shu

Electrochemical Luminescent DNA Sensor Based on Polymerase-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 June 2017
Revised Date: 10 November 2017

Fund Project: This work was supported by the National Natural Science Foundation of China (Nos. 21375031, 21765006), the Natural Science Foundation of Guangxi Province, China (No. 2015GXNSFFA139005), and the High Level Innovation Teams of Guangxi Colleges & Universities and Outstanding Scholars Program of China (Gui Jiao Ren[2014] No. 49).

A novel polymerase-based electrochemiluminescence DNA sensor was constructed for messenger RNA (mRNA) detection by cyclic chain displacement polymerization, assisted by target mRNA cycle, and quantum dots signal amplification. Firstly, the mercapto-modified capture-type capture DNA (CP) was immobilized on the surface of a magneto-controlled glassy carbon electrode via Au-S bond. After adding the target mRNA, CP was opened and hybridized with mRNA to form dsDNA. After adding polymerase, primer chain (DNA1) and the base, the primer chain was extended to replace the target mRNA. After one cycle, the mRNA chain could open another hairpin in order to carry out next cycle of amplification. Finally, electrochemical luminescence detection was carried out by adding DNA2 labeled TGA-CdTe quantum dots. The amplification of the target mRNA by the addition of polymerase and the signal combined with the quantum dot mark improved the sensitivity of the sensor greatly. The result showed that the logarithm of target mRNA concentration had a good linear relationship with the corresponding ECL signal in the range of 1×10^-15-1×10^-11 mol/L, with the detection limit of 3.4×10^-16 mol/L(S/N=3). Under the optimal conditions, the recoveries of mRNA spiked in human serum sample were 97.2%-102.3%. This sensor exhibited good selectivity, stability and reproducibility.
- DNA sensor,
- Polymerase,
- Target cycle,
- Signal amplification,
- Electrochemical luminescence
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