NDLI: Quadruple signal amplification strategy based on hybridization chain reaction and an immunoelectrode modified with graphene sheets, a hemin/G-quadruplex DNAzyme concatamer, and alcohol dehydrogenase: ultrasensitive determination of influenza virus subtype H7N9

Content Provider	Springer Nature Link
Author	Yang, Juan Xiang, Yu Song, Chao Liu, Lingzhi Jing, Xiaoying Xie, Guoming Xiang, Hua
Copyright Year	2015
Abstract	We report on a new amplification strategy for use in an immunoassay for influenza virus subtype H7N9. Graphene sheets were first placed on a glassy carbon electrode (GCE), and gold nanoparticles were then electrodeposited as a support for a layer of alcohol dehydrogenase (ADH) in a sol–gel containing thiol groups. Protein A was used to properly orientate immobilized antibody against H7N9 on the sol–gel, and this is shown to result in strongly improved specificity of the antigen-antibody binding. Thus, a sensitive and specific immunosensor was obtained in which a quadruple signal amplification strategy is employed, viz. (a) via the use of graphene sheets, (b) via a hybridization chain reaction, (c) the use of hemin/G-quadruplex DNAzyme concatamers, and (d) the use of ADH. The hemin/G-quadruplex is a typical DNAzyme, which simultaneously acts as NADH oxidase and HRP-mimicking DNAzyme. The hybridization chain reaction-based DNAzyme concatamers assembled on multi-walled carbon nanotubes (MWCNTs) and the ADH represent a triple electrocatalytic enzyme cascade system. Sandwich immunoreactions occurred between the capture antibody on the electrode and the secondary antibody labeled with MWCNTs. Positively charged Methylene Blue (MB) was then used as an intercalator to detect the DNAzyme concatamer formed. The differential pulse voltammetric signals for MB are related to the concentration of H7N9 in the range from 8 to 60 pg · mL−1, and the detection limit is 0.81 pg · mL−1 (at an S/N ratio of 3). This immunoassay is very sensitive, specific and robust. Graphical Abstract An electrochemical sandwich immunosensor has been developed for sensitive and specific detection of influenza virus subtype H7N9. Protein A was used to properly orientate antibody. The hybridization chain reaction based DNAzyme concatamers assembled on multi-walled carbon nanotubes (MWCNTs) and the ADH represent a triple electrocatalytic enzyme cascade system.
Starting Page	2377
Ending Page	2385
Page Count	9
File Format	PDF
ISSN	00263672
Journal	Microchimica Acta
Volume Number	182
Issue Number	15-16
e-ISSN	14365073
Language	English
Publisher	Springer Vienna
Publisher Date	2015-08-05
Publisher Place	Vienna
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	H7N9 Oriented antibody binding Quadruple signal amplification Enzyme-catalysis Immunosensor Multiwalled carbon nanotubes Influenza Nanochemistry Nanotechnology Characterization and Evaluation of Materials Analytical Chemistry Microengineering
Content Type	Text
Resource Type	Article
Subject	Analytical Chemistry

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