DEVELOPMENT AND STANDARDIZATION OF AN INDIRECT ELISA FOR THE DIAGNOSIS MAEDI-VISNA IN SHEEP

Authors

  • Tânia Valeska Medeiros Dantas Universidade Estadual do Ceará (UECE), Fortaleza, Ceará, Brasil
  • Suzana Aparecida Costa de Araújo Universidade Estadual do Ceará (UECE), Fortaleza, Ceará, Brasil
  • Raymundo Rizaldo Pinheiro CNPC/EMBRAPA
  • Maria Alzira do Carmo Aragão Universidade Estadual do Ceará (UECE), Fortaleza, Ceará, Brasil
  • Jean Berg Alves da Silva Universidade Estadual do Ceará (UECE), Fortaleza, Ceará, Brasil
  • Aracely Rafaelle Fernandes Ricarte Universidade Estadual do Ceará (UECE), Fortaleza, Ceará, Brasil
  • Adriana Lopes Ribeiro Universidade Estadual do Ceará (UECE), Fortaleza, Ceará, Brasil
  • Maria Fátima da Silva Teixeira Universidade Estadual do Ceará (UECE), Fortaleza, Ceará, Brasil

Keywords:

Medicina Veterinária Preventiva

Abstract

The objective of this work was to develop and standardize an indirect ELISA for diagnosis of Maedi Visna (MV) infection. The antigen was produced from the supernatants of caprine synovial membrane (CSM) cell monolayers inoculated with Maedi Visna virus (MVV), strain K1514, by cycles of freezing and thawed, and clarified by centrifugation at 3000 g for 40 min. The clarified suspension was precipitated with PEG 8000 and centrifugated at 12000 g for 60 min; the pellet was resuspended in buffer TNE and layered onto a sucrose cushion by centrifugation at 42000 g for 105 min. The pellet was resuspended in PBS containing phenylmethylsulphonyl fluoride (PMSF). The ELISA was performed in 96 wells plates, incubated for 1 h at 37 ºC. The reaction was detected by incubation with the enzyme substrate and o-phenylenediamine (OPD) during 15 min.  The tests were compared using 175 sera samples. The concentration of the antigen was 2 µg/mL and the best dilution of the sera was 1:100. ELISA detected more positive samples (22.9%) than AGID (6.3%) and presented a better sensitivity than AGID and, although the specificity was below the expected, it could already be recomended in the diagnosis of MVV infection.KEY WORDS: ELISA, MaediVisna, serologic diagnosis, sheep lentivirus.

Downloads

Download data is not yet available.

Downloads

Published

2008-04-03

How to Cite

DANTAS, Tânia Valeska Medeiros; ARAÚJO, Suzana Aparecida Costa de; PINHEIRO, Raymundo Rizaldo; ARAGÃO, Maria Alzira do Carmo; SILVA, Jean Berg Alves da; RICARTE, Aracely Rafaelle Fernandes; RIBEIRO, Adriana Lopes; TEIXEIRA, Maria Fátima da Silva. DEVELOPMENT AND STANDARDIZATION OF AN INDIRECT ELISA FOR THE DIAGNOSIS MAEDI-VISNA IN SHEEP. Brazilian Animal Science/ Ciência Animal Brasileira, Goiânia, v. 9, n. 1, p. 181–187, 2008. Disponível em: https://revistas.ufg.br/vet/article/view/3677. Acesso em: 6 dec. 2025.

Issue

Vol. 9 No. 1 (2008)

Section

Veterinary Medicine

License

Copyright (c) 2008 Brazilian Animal Science/ Ciência Animal Brasileira

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Authors who publish with this journal agree to the following terms:

  1. Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
  2. Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
  3. Authors are permitted and encouraged to post their work online (e.g. in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).