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Immunobiological for the laboratory diagnosis of ranavirose: cloning and expression of the MCP gene of Brazilian Ranavirus isolate and production of polyclonal antibodies against recombinant MCP protein

Grant number: 17/01356-4
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): July 01, 2017
Effective date (End): June 30, 2019
Field of knowledge:Agronomical Sciences - Veterinary Medicine
Cooperation agreement: Coordination of Improvement of Higher Education Personnel (CAPES)
Principal Investigator:Ricardo Luiz Moro de Sousa
Grantee:Thaís Camilo Corrêa
Home Institution: Faculdade de Zootecnia e Engenharia de Alimentos (FZEA). Universidade de São Paulo (USP). Pirassununga , SP, Brazil

Abstract

The genus Ranavirus is responsible for acute or systemic diseases in fish, amphibians and reptiles, including species of zootechnical interest, causing high mortality and morbidity, and therefore generate large ecological and economic impacts in several parts of the world. The best studied gene in Ranavirus is that encoding the major capsid protein (MCP), a 50 kDa structural protein with highly conserved domains comprising 40-45% of the virus protein structure. The present project has as objectives: produce the recombinant MCP protein (rMCP); producing polyclonal anti-rMCP antibodies; and to standardize an Indirect-Blocking ELISA (ELISA-IB) test to detect the presence of anti-ranavirus antibodies in fish serum. In order to do so, the Brazilian isolate of Ranavirus, Frog virus 3 (FV3-simile belonging to the Laboratório de Higiene Zootécnica of FZEA/USP), will be used for the cloning and expression of the MCP gene in the prokaryotic expression system (Escherichia coli), serum polyclonal antibodies will be produced from experimental animals (rabbits), and the ELISA-IB test will be standardized according to FAO/OIE recommendations. In view of the scarcity of information on the occurrence of ranavirosis in fish in Brazil, the importance of aquaculture for Brazilian zootechnical production and possible economic impact caused by infectious diseases of the genus Ranavirus, this project is intends to fill in the absence existing in the country in the area of monitoring and epidemiology of viral diseases of fish, due to the lack of immunoreagents necessary to carry out specific laboratory tests for viral variants found in Brazil. (AU)