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Development of recombinant NS1-NS3 antigen based indirect ELISA for detection of bluetongue antibodies in sheep.

Abstract
Bluetongue is an insect borne (Culicoides) viral disease of small ruminants. The virus blankets the globe with a wide serotypic variation, numbered from 1 to 28. In India 21 different serotypes have been reported to be circulating across the various agro-climatic zones of the country. Non-structural proteins (NSPs) of bluetongue virus have always remained ideal target for differentiation of infected from vaccinated animals. The current study is an extrapolation of our previous work where a novel fusion construct comprising of bluetongue viral segment NS1 and NS3 was successfully cloned, expressed, purified with an efficient strategy for its suitable implementation as a diagnostic antigen. In this study, the applicability of the fusion construct has been further evaluated and optimised for field applicability. The fusion construct used in an ELISA platform projected a relative diagnostic sensitivity and specificity of 98.1% and 95.5% respectively against a pre-established test panel. The rNS1-NS3 ELISA showed substantially good agreement with the commercial BTV antibody detection kit. Finally, the study brings together the diagnostic capability of two NSPs, which can be a handy tool for sero-surveillance of bluetongue.
AuthorsNihar Nalini Mohanty, Divakar Hemadri, Archana Munivenkatarayappa, Namrata Shetty, Vinutha Subramanyam, Sanchay Kumar Biswas, Mohammed Mudassar Chanda, Sathish B Shivachandra
JournalJournal of immunological methods (J Immunol Methods) Vol. 490 Pg. 112959 (03 2021) ISSN: 1872-7905 [Electronic] Netherlands
PMID33412173 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
CopyrightCopyright © 2021 Elsevier B.V. All rights reserved.
Chemical References
  • Antibodies, Viral
  • NS1 protein, bluetongue virus
  • Recombinant Fusion Proteins
  • S10 protein, Bluetongue virus
  • Viral Nonstructural Proteins
Topics
  • Animals
  • Antibodies, Viral (blood)
  • Bluetongue (diagnosis, immunology)
  • Bluetongue virus (physiology)
  • Enzyme-Linked Immunosorbent Assay (methods)
  • Immunity, Humoral
  • Recombinant Fusion Proteins (genetics, metabolism)
  • Sheep (immunology, virology)
  • Viral Nonstructural Proteins (genetics, metabolism)

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