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A Nano-Au/C-MWCNT based label free amperometric immunosensor for the detection of capsicum chlorosis virus in bell pepper

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Abstract

Accurate and on time diagnosis of plant viruses is an essential prerequisite for efficient control in field conditions. A number of diagnostic methods have been reported with the required level of sensitivity. Here, we propose a label free immunosensor for efficient and sensitive detection of capsicum chlorosis virus (CaCV) in bell pepper. Antigen was immobilized over the surface of gold nanoparticle/multi-walled carbon nanotube (Nano-Au/C-MWCNT) screen printed electrodes using 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC)/N-hydroxysuccinimide (NHS) cross linking chemistry followed by interaction with groundnut bud necrosis virus (GBNV)/CaCV specific polyclonal antibody. The electrochemical response was measured by cyclic voltammetry (CV), differential pulse voltammetry (DPV) using the redox indicator. Electrode surface characterization was done by performing scanning electron microscopy (SEM). Electrochemical studies showed positive results at different antigenic dilutions ranging from 10−2 – 8x10−5. The sensitivity of the immunosensor developed has been compared with direct antigen coated enzyme-linked immunosorbent assay (DAC-ELISA) and the results showed that the immunosensor developed was 800-1000 times more sensitive, when compared to DAC-ELISA for CaCV detection. The immunosensor we have developed is economical and sensitive and could be used for immediate determination of the presence of virus in extracts from bell pepper leaves.

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Acknowledgements

The authors would like to thank Prof. P. K. Khosla, Hon’ble Vice-Chancellor, Shoolini University of Biotechnology and Management Sciences, Solan and Foundation for Life Sciences and Business Management (FLSBM), Solan for providing financial support and the necessary facilities. Authors are also thankful to Dr Bikash Mandal (Senior Scientist), Indian Agricultural Research Institute (IARI) for kindly providing CaCV/GBNV-specific antibodies.

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Authors and Affiliations

  1. Faculty of Applied Sciences and Biotechnology, Shoolini University of Biotechnology and Management Sciences, Bajhol, Solan, Himachal Pradesh, 173229, India

    Anshul Sharma, Ankur Kaushal & Saurabh Kulshrestha

  2. Department of Food Science and Biotechnology, Gachon University, Seongnam, 13120, South Korea

    Anshul Sharma

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  1. Anshul Sharma
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  2. Ankur Kaushal
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  3. Saurabh Kulshrestha
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Correspondence to Saurabh Kulshrestha.

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Sharma, A., Kaushal, A. & Kulshrestha, S. A Nano-Au/C-MWCNT based label free amperometric immunosensor for the detection of capsicum chlorosis virus in bell pepper. Arch Virol 162, 2047–2052 (2017). https://doi.org/10.1007/s00705-017-3293-5

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  • DOI: https://doi.org/10.1007/s00705-017-3293-5

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