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Detection of bluetongue virus group-specific antigen using monoclonal antibody based sandwich ELISA

Virologica Sinica volume 25, pages 390–400 (2010)Cite this article

Abstract

A monoclonal antibody (MAb) specific for the bluetongue virus (BTV) group specific antigen (VP7) was characterized for its reactivity with purified virus and recombinant BTV VP7 (rVP7) protein and its suitability for use in the sandwich ELISA. The MAb, designated as 5B5 was specific to VP7 and belongs to IgG2a subclass and was selected for the development of the sELISA in this study. The MAb had a titer of 1:25 with BTV and 1:2 with the rVP7 protein. The sELISA is based on capturing of BTV antigen with VP7 specific MAb followed by detection using BTV polyclonal antiserum raised in rabbits. The assay was evaluated with six cell culture adapted serotypes of BTV that have been isolated from India, 1, 2, 15, 17, 18 and 23. The assay could detect BTV antigen as early as day 8 in blood. It was also successfully applied for the detection of BTV group specific antigen in clinical samples of blood, washed RBCs, buffy coat and plasma. A total of 102 field samples from animals, suspected of being infected with BTV, were tested and 29.42% were positive. The blood samples were also amplified in cell culture which improved the sensitivity of the assay. Results confirmed that the sELISA is rapid and specific.

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Authors and Affiliations

  1. Division of Virology, Indian Veterinary Research Institute, Mukteswar, 263138, Nainital (Distt.), Uttarakhand, India

    Pradeep Narayan Gandhale, Veerakyathappa Bhanuprakash & Gnanavel Venkatesan

  2. Project Directorate on Animal Disease Monitoring And Surveillance, 560024, Bangalore, India

    Vinayagamurthy Balamurugan

  3. Indian Veterinary Research Institute, Campus Bangalore, 560024, India

    Madhusudhan Hosamani

  4. National Research Centre on Equines, Hisar, 125001, Haryana, India

    Raj Kumar Singh

Authors
  1. Pradeep Narayan Gandhale
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  2. Veerakyathappa Bhanuprakash
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  3. Vinayagamurthy Balamurugan
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  4. Madhusudhan Hosamani
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  5. Gnanavel Venkatesan
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  6. Raj Kumar Singh
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Correspondence to Veerakyathappa Bhanuprakash.

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Gandhale, P.N., Bhanuprakash, V., Balamurugan, V. et al. Detection of bluetongue virus group-specific antigen using monoclonal antibody based sandwich ELISA. Virol. Sin. 25, 390–400 (2010). https://doi.org/10.1007/s12250-010-3160-y

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  • DOI: https://doi.org/10.1007/s12250-010-3160-y

Key words

  • Bluetongue
  • Diagnosis
  • ELISA
  • Monoclonal antibody
  • Sandwich ELISA