Advances in Animal and Veterinary Sciences

Research Article
Adv. Anim. Vet. Sci. 8(8): 853-860
Http://dx.doi.org/10.17582/journal.aavs/2020/8.8.853.860
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Barkha Ratta1, Swagatika Priyadarsini1, Nikhil K. Channabasappa1, Pashupathi Mani1, Meeta Saxena1, R Saravanan2, Ajay Kumar1*

1Division of Biochemistry, Indian Veterinary Research Institute, Izatnagar, Bareilly (UP), 243122, India; 2Immunology Section, Indian Veterinary Research Institute, Izatnagar, Bareilly (UP), 243122, India.

Abstract | Bovine herpesvirus 1 (BoHV-1) affects mainly large ruminants (cattle and buffalo), resulting in a substantial negative impact on the industry. The virus causes respiratory symptoms, abortion and a decrease in milk production. Among envelope proteins of the virus, the glycoprotein D (gD) is considered a dominant immunogenic. We expressed the immunogenic portion of gD in the prokaryotic system (E. cloni 10G) and did evaluate its potential as a diagnostic antigen for indirect ELISA. Using the Ni-HRP probe, the expressed recombinant gD protein (rgD, 45kDa) was characterized by SDS-PAGE and western blotting. The immunogenic reactivity of red was optimized for indirect ELISA using known true positive and negative sera (antibodies) of the virus. Sera (n=158) were screened and, with a cut-off value of 0.238, relative diagnostic sensitivity (69%) and specificity (52.3%) of in-house developed indirect ELISA was determined. With these preliminary results, the study suggests the diagnostic potential of recombinant gD protein for serodiagnosis of BoHV-1 infection at a large scale.

Keywords | Indirect ELISA, Bovine herpesvirus-1, Glycoprotein D, Antibodies, Recombinant.