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Publications

Journal Article Ppublish

PubMed: 39824477 DOI: 10.1016/j.jim.2025.113801

Expression of bluetongue virus full-length VP7 protein in insect cells and its diagnostic utility for detection of antibodies to the virus infection.

J Immunol Methods 538:113801

Authors: Sanchay Kumar Biswas, Madhusudan Hosamani, Karam Chand, Ankita Chauhan, Kurat Ul Ain, Vanitha Selvarajan, Sushmita Nautiyal, Muzamil Bashir, Divakar Hemadri, Gaurav Kumar Sharma, B P Sreenivasa

Affiliations

  • ICAR-Indian Veterinary Research Institute, Centre for Animal Disease Research and Diagnosis, Izatnagar, Bareilly 243 122, UP, India.
  • ICAR-Indian Veterinary Research Institute, Bangalore, Karnataka 560024, India.
  • ICAR-Indian Veterinary Research Institute, Mukteswar, Kumaon, Uttarakhand 263138, India.
  • ICAR-Indian Veterinary Research Institute, Centre for Animal Disease Research and Diagnosis, Izatnagar, Bareilly 243 122, UP, India.
  • ICAR-National Institute of Veterinary Epidemiology and Disease Informatics, Bangalore, 560064, India.

Abstract

Bluetongue (BT) is a vector-borne viral disease of multiple domestic and wild ruminants across the globe. The VP7 protein of bluetongue virus (BTV) is the major immune-dominant structural protein that is conserved across the BTV serotypes and therefore, targeted for the development of immuno-diagnostics for BT. In this study, full-length recombinant VP7 protein (rVP7) of BTV-1 was expressed in Trochoplusia ni derived insect cells (Tn5) using codon-optimized synthetic gene construct through baculovirus expression system. The seed stock of recombinant baculovirus was amplified to a high titre (>1 × 10 pfu/ml) at P2 and P3 in sf9 cells. The rVP7 was successfully produced and purified from infected Tn5 culture lysate for evaluation of the immuno-reactivity and its diagnostic potential. The purified protein showed strong reactivity in western blot analysis with the polyclonal immune serum produced against BTV core antigen in guinea pigs. An indirect ELISA (iELISA) was optimized by using the purified rVP7 for the detection of the group-specific antibodies to BTV in sheep and goats. The iELISA was found to be highly sensitive (98.9 %), specific (98.1 %), and reproducible (CV < 10 %) for detection of the antibodies to BTV in sheep and goat serum. The iELISA could detect the specific antibodies in naturally infected goat serum containing type-specific neutralizing antibodies to different BTV serotypes indicating the potential of the rVP7 for the development of the group-specific sero-diagnostics for BT.

MeSH

Cells, Bluetongue, Serum, Antibodies, Baculoviridae, Viruses, Bluetongue Virus, Infections, Virus Diseases

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