A label-free electrochemical assay for coronavirus IBV H120 strain quantification based on equivalent substitution effec
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ORIGINAL PAPER
A label-free electrochemical assay for coronavirus IBV H120 strain quantification based on equivalent substitution effect and AuNPs-assisted signal amplification Yazhi Yang 1 & Dawei Yang 2 & Yingge Shao 1 & Yi Li 1 & Xifeng Chen 2 & Yuanyuan Xu 1
&
Jinfeng Miao 1
Received: 2 June 2020 / Accepted: 29 September 2020 # Springer-Verlag GmbH Austria, part of Springer Nature 2020
Abstract 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 μM with the detection limit of 2.96e−10 μM 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.
Keywords Coronavirus . IBV H120 strain . Equivalent substitution effect . AuNPs-assisted signal amplification . Electrochemical assay
Introduction
Yazhi Yang and Dawei Yang contributed equally to this work. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00604-020-04582-3) contains supplementary material, which is available to authorized users. * Yuanyuan Xu [email protected] * Jinfeng Miao [email protected] 1
MOE Joint International Research Laboratory of Animal Health and Food Safety, Key Laboratory of Animal Physiology & Biochemistry, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China
2
Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou 215163, China
In the last few decades, viruses are a real menace to safety. The pandemic dimension spread of coronavirus disease poses a severe threat to the health and lives of seven billion people worldwide [1]. Rapid identification of viruses should be one of the best ways to prevent disease outbreaks and is of great significance to medical healthcare [1]. IBV, one kind of coronaviruses, is a positive-sense single-stranded enveloped RNA virus with a length of 27–
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