The rGO@AuNPs modified label-free electrochemical immunosensor to sensitive detection of CP-BNYVV protein of Rhizomania disease agent in sugar beet.

IF 4.7 2区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS
Marziye Karimzade, Hashem Kazemzadeh-Beneh, Negar Heidari, Mehrasa Rahimi Boroumand, Parviz Norouzi, Mohammad Reza Safarnejad, Masoud Shams-Bakhsh
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Abstract

For the first time, a novel simple label-free electrochemical immunosensor was fabricated for sensitive detection of the coat protein of beet necrotic yellow vein virus (CP-BNYVV) as the causal agent of Rhizomania disease in sugar beet. To boost the amplification of the electrochemical signal, gold nanoparticles-reduced graphene oxide (AuNPs-rGO) nanocomposite was employed to modify the glassy carbon electrode. Anti-BNYVV polyclonal was immobilized onto a modified electrode by applying a thiol linker via a self-assembly monolayer (SAM) and activating the functionalized surface using (3-aminopropyl triethoxysilane) and glutaraldehyde. The determination step relied on the forming of an immunocomplex between the antigen and oriented antibody, resulting in a decrease in current in the [Fe (CN)6]3-/4- redox reaction. The response value exhibited direct proportionality to the concentrations of CP-BNYVV. Scanning electron microscopy, energy dispersive x-ray, cyclic voltammetry, and electrochemical impedance spectroscopy techniques collectively provided a comprehensive understanding of the structural, morphological, and electrochemical features during the modification steps. Under optimized experimental conditions, the fast Fourier transform square wave voltammetry responds to the logarithm of CP-BNYVV concentrations in a wide linear range from 0.5 to 50000 pg/mL and the limit of detection is calculated to be 150 fg/mL, implying the admirable sensitivity. Selectivity assay exhibited no cross-reactivity with other proteins from interfering virus samples. Satisfactory reproducibility and stability were achieved with a relative standard deviation of 3.1% and a stable value of 90% after 25 days, respectively. More importantly, the high performance of the immunosensor resulted in the direct detection of CP-BNYVV in spiked and infected plant samples, which affords a sensing platform with huge potential application for the early detection of BNYVV virus in field conditions.

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来源期刊
Plant Methods
Plant Methods 生物-植物科学
CiteScore
9.20
自引率
3.90%
发文量
121
审稿时长
2 months
期刊介绍: Plant Methods is an open access, peer-reviewed, online journal for the plant research community that encompasses all aspects of technological innovation in the plant sciences. There is no doubt that we have entered an exciting new era in plant biology. The completion of the Arabidopsis genome sequence, and the rapid progress being made in other plant genomics projects are providing unparalleled opportunities for progress in all areas of plant science. Nevertheless, enormous challenges lie ahead if we are to understand the function of every gene in the genome, and how the individual parts work together to make the whole organism. Achieving these goals will require an unprecedented collaborative effort, combining high-throughput, system-wide technologies with more focused approaches that integrate traditional disciplines such as cell biology, biochemistry and molecular genetics. Technological innovation is probably the most important catalyst for progress in any scientific discipline. Plant Methods’ goal is to stimulate the development and adoption of new and improved techniques and research tools and, where appropriate, to promote consistency of methodologies for better integration of data from different laboratories.
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