Produção da proteína recombinante p26 fusionada à MBP para diagnóstico da anemia infecciosa equina

• Ano de defesa: 2017
• Autor(a) principal: FONTES, Karin Florencio Lins de Paiva
• Orientador(a): CASTRO, Roberto Soares de
• Banca de defesa: SOUZA, Paulo Roberto Eleutério de, RIZZO, Huber, JESUS, André Luiz Santos de
• Tipo de documento: Tese
• Tipo de acesso: Acesso aberto
• dARK ID: ark:/57462/0013000008c12
• Idioma: por
• Instituição de defesa: Universidade Federal Rural de Pernambuco
• Programa de Pós-Graduação: Programa de Pós-Graduação em Ciência Veterinária
• Departamento: Departamento de Medicina Veterinária
• País: Brasil
• Palavras-chave em Português: Anemia Infecciosa; Equino; Proteína recombinante; Diagnóstico
• Área do conhecimento CNPq: CIENCIAS AGRARIAS::MEDICINA VETERINARIA
• Link de acesso: http://www.tede2.ufrpe.br:8080/tede2/handle/tede2/7105

Equine Infectious Anemia (EIA) is caused by a lentivirus of the family Retroviridae and is considered one of the most important viruses in horses in the world. It is a chronic infection, untreated, prevalent in hot and humid climate regions favorable to transmission by hematophagous insects. The official diagnosis of the disease is made through the detection of circulating antibodies in the IDGA and ELISA tests. In order to increase protein yield and improve the performance of serological tests, the p26 gene sequence was optimized for preferred codons for use in E. coli and fused to maltose binding protein. The resulting recombinant fusion protein (p26rec) was detected by SDS-PAGE and Western blot with anti- HIS monoclonal antibody and used as an antigen in the development and evaluation to evaluate an IDGA and an ELISA (IDGArec and ELISArec) for the diagnosis of EIA. After analysis of 569 equine sera, diagnostic sensitivity (SeD) and diagnostic specificity (SpD) were determined. The cutoff point for ELISArec (> 29.51%) was determined by the ROC curve analysis. For IDGArec both SeD and SpD were 100% and for ELISArec were 100% and 99.6% respectively. The p26rec has been maintained with stable working dilutions for more than three years stored at 4 ° C. The results demonstrated the high degree of confidence obtained with p26rec in the IDGArec and ELISArec tests and could therefore be recommended in the serological diagnosis of the AIE.