Ultrasensitive Electrochemical DNA Biosensor Fabrication by Coupling an Integral Multifunctional Zirconia-Reduced Graphene Oxide-Thionine Nanocomposite and Exonuclease I-Assisted Cleavage

被引:19
作者
Chen, Zhiqiang [1 ]
Liu, Xueqian [2 ]
Liu, Dengren [1 ]
Li, Fang [2 ]
Wang, Li [1 ]
Liu, Shufeng [1 ]
机构
[1] Qingdao Univ Sci & Technol, Coll Chem & Mol Engn, Minist Educ, Key Lab Opt Elect Sensing & Analyt Chem Life Sci, Qingdao, Peoples R China
[2] Qingdao Univ Sci & Technol, Coll Marine Sci & Biol Engn, Qingdao, Peoples R China
基金
中国国家自然科学基金;
关键词
electrochemical DNA biosensor; reduced graphene oxide; zirconia; signal amplification; exonuclease I; STRAND-DISPLACEMENT; AMPLIFICATION STRATEGY; CASCADE AMPLIFICATION; SIGNAL AMPLIFICATION; ENDONUCLEASE; PROTEIN; NANOCLUSTERS; MICROARRAYS; BIOMARKERS; BEACON;
D O I
10.3389/fchem.2020.00521
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
In this work, a simple but sensitive electrochemical DNA biosensor for nucleic acid detection was developed by taking advantage of exonuclease (Exo) I-assisted cleavage for background reduction and zirconia-reduced graphene oxide-thionine (ZrO2-rGO-Thi) nanocomposite for integral DNA recognition, signal amplification, and reporting. The ZrO2-rGO nanocomposite was obtained by a one-step hydrothermal synthesis method. Then, thionine was adsorbed onto the rGO surface,via pi-pi stacking, as an excellent electrochemical probe. The biosensor fabrication is very simple, with probe DNA immobilization and hybridization recognition with the target nucleic acid. Then, the ZrO2-rGO-Thi nanocomposite was captured onto an electrodeviathe multicoordinative interaction of ZrO(2)with the phosphate group on the DNA skeleton. The adsorbed abundant thionine molecules onto the ZrO2-rGO nanocomposite facilitated an amplified electrochemical response related with the target DNA. Since upon the interaction of the ZrO2-rGO-Thi nanocomposite with the probe DNA an immobilized electrode may also occur, an Exo I-assisted cleavage was combined to remove the unhybridized probe DNA for background reduction. With the current proposed strategy, the target DNA related with P53 gene could be sensitively assayed, with a wide linear detection range from 100 fM to 10 nM and an attractive low detection limit of 24 fM. Also, the developed DNA biosensor could differentiate the mismatched targets from complementary target DNA. Therefore, it offers a simple but effective biosensor fabrication strategy and is anticipated to show potential for applications in bioanalysis and medical diagnosis.
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页数:11
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