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Efficient immunodiagnosis of Citrus yellow mosaic virus using polyclonal antibodies with an expressed recombinant virion-associated protein

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Abstract

Citrus yellow mosaic virus (CYMV) is a member of genus Badnavirus of the family Caulimoviridae. It is the causal agent of citrus yellow mosaic disease in citrus and causes reduction in yield. As the virus is vegetative propagated by grafting, development of high-throughput diagnosis methods based on serological techniques is a prerequisite for production of healthy virus-free planting material. The current study describes the development of polyclonal antibodies raised in rabbits against purified recombinant virion-associated protein (rVAP) encoded by ORF-II of CYMV. The specificity of developed antiserum was evaluated in immunosorbent electron microscopy (ISEM), antigen-coated plate-enzyme linked immunosorbent assay (ACP-ELISA) and immunocapture PCR (IC-PCR). The antiserum specifically reacted up to a dilution of 1:2000 in ACP-ELISA for detection of CYMV-infected plants. The antiserum was validated by screening CYMV-infected plants maintained in the glass house through ACP-ELISA. To the best for our knowledge, this is the first report on production of polyclonal antiserum using recombinant virion-associated protein as fusion protein, which could be used for screening CYMV-infected plants by ELISA and IC-PCR. These immunodiagnostic methods can be effectively employed in routine indexing of citrus and in quarantine process.

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Acknowledgements

We thank ICAR-Outreach project (Grant no. 02) to bear financial assistance for carrying out experiments, Head, Division of Plant Pathology and Director, ICAR-Indian Agricultural Research Institute, New Delhi for providing necessary lab facilities.

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Authors and Affiliations

  1. Division of Plant Pathology, Advanced Centre for Plant Virology, ICAR-Indian Agricultural Research Institute, New Delhi, 110012, India

    P. Vignesh Kumar & Virendra Kumar Baranwal

  2. ICAR Research Complex for NEH Region, Manipur Centre, Imphal, 795004, India

    Susheel Kumar Sharma

  3. Amity Institute of Virology and Immunology, Amity University, Noida, 201303, India

    Narayan Rishi

Authors
  1. P. Vignesh Kumar
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  2. Susheel Kumar Sharma
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  3. Narayan Rishi
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  4. Virendra Kumar Baranwal
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Correspondence to Virendra Kumar Baranwal.

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Kumar, P.V., Sharma, S.K., Rishi, N. et al. Efficient immunodiagnosis of Citrus yellow mosaic virus using polyclonal antibodies with an expressed recombinant virion-associated protein. 3 Biotech 8, 39 (2018). https://doi.org/10.1007/s13205-017-1063-4

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