期刊
MICROCHIMICA ACTA
卷 187, 期 11, 页码 -出版社
SPRINGER WIEN
DOI: 10.1007/s00604-020-04582-3
关键词
Coronavirus; IBV H120 strain; Equivalent substitution effect; AuNPs-assisted signal amplification; Electrochemical assay
资金
- Natural Science Foundation of Jiangsu Province for Excellent Young Scholars [BK20170087]
- Fundamental Research Funds for the Central Universities [KYGD202001]
- Key Project of Inter-governmental International Scient i f i c and Technological Innovation Cooperation [2018YFE0102200]
- Key Scientific and Technological Project of XPCC [2020AB025]
- Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD)
A label-free electrochemical strategy is proposed combining equivalent substitution effect with AuNPs-assisted signal amplification. According to the differences of S1 protein in various infectious bronchitis virus (IBV) strains, a target DNA sequence that can specifically recognize H120 RNA forming a DNA-RNA hybridized double-strand structure has been designed. Then, the residual single-stranded target DNA is hydrolyzed by S1 nuclease. Therefore, the content of target DNA becomes equal to the content of virus RNA. After equivalent coronavirus, the target DNA is separated from DNA-RNA hybridized double strand by heating, which can partly hybridize with probe 2 modified on the electrode surface and probe 1 on AuNPs' surface. Thus, AuNPs are pulled to the surface of the electrode and the abundant DNA on AuNPs' surface could adsorb a large amount of hexaammineruthenium (III) chloride (RuHex) molecules, which produce a remarkably amplified electrochemical response. The voltammetric signal of RuHex with a peak near - 0.28 V vs. Ag/AgCl is used as the signal output. The proposed method shows a detection range of 1.56e(-9) to 1.56e(-6) mu M with the detection limit of 2.96e(-10) mu M for IBV H120 strain selective quantification detection, exhibiting good accuracy, stability, and simplicity, which shows a great potential for IBV detection in vaccine research and avian infectious bronchitis diagnosis.
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