Cloning, expression and immunological evaluation of the Omp31 protein of Brucella melitensis and evaluation of its possible use for the diagnosis in bovine brucellosis

Authors

  • Jose David Rosales Rangel Centro de Inmunoproducción Masiva de Anticuerpos, Fundación Instituto de Estudios Avanzados (IDEA), Caracas
  • Annie Castillo Ochoa Centro de Inmunoproducción Masiva de Anticuerpos, Fundación Instituto de Estudios Avanzados (IDEA), Caracas
  • Armando Reyna Bello Carrera Ingeniería en Biotecnología, Departamento de Ciencias de la Vida, Universidad de las Fuerzas Armadas ESPE, Santo Domingo
  • Ana Teresa Serrano Facultad de Ciencias Veterinarias, Universidad Central de Venezuela, Maracay
  • Rodolfo Fernandez Gomez Carrera Ingeniería en Biotecnología, Departamento de Ciencias de la Vida, Universidad de las Fuerzas Armadas ESPE, Sangolquí

DOI:

https://doi.org/10.15381/rivep.v29i3.14008

Keywords:

brucellosis, Omp31r, recombinant protein

Abstract

The serological diagnosis of brucellosis is carried out by the detection of antibodies generated against the liposaccharide «LPS» or bacterial extracts of whole cells by ELISA or agglutination tests. The objective of this study was to evaluate a recombinant protein that can be used in the serological diagnosis of bovine brucellosis. The Omp31 gene of Brucella melitensis 16M was chosen. The gene was cloned from DNA of B. melitensis; the expression of the Omp31r protein was performed in a prokaryotic expression system using the pET28a vector and purification by metal affinity chromatography (IMAC). An indirect ELISA was standardized using control sera from positive and negative bovines. The Omp31r protein reacted with the positive control sera and succeeded in discriminating against the negative sera. The sensitivity and specificity of the ELISA assay was 77.2% and 90.6%, respectively. This indirect ELISA system can be useful for the rapid diagnosis of large numbers of samples in the diagnosis of bovine brucellosis.

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Author Biography

  • Jose David Rosales Rangel, Centro de Inmunoproducción Masiva de Anticuerpos, Fundación Instituto de Estudios Avanzados (IDEA), Caracas

    Investigador, Area de Salud.Centro de Inmunoproduccin de Anticuerpos y Unidad de Pesquisa Neonatal.

    Busqueda de antigenos y anticuerpos recombinantes "NanoAnticuerpos" para el desarrollo de test de diagnostico en enfermedades humanas, veteinarias y agricolas.

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Published

2018-09-06

Issue

Vol. 29 No. 3 (2018)

Section

Artículos Primarios

License

Copyright (c) 2018 Jose David Rosales Rangel

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

AUTHORS RETAIN THEIR RIGHTS:

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b. Authors retain their right to share, copy, distribute, perform and publicly communicate their article (eg, to place their article in an institutional repository or publish it in a book), with an acknowledgment of its initial publication in the Revista de Investigaciones Veterinarias del Perú (RIVEP).

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How to Cite

Rosales Rangel, J. D., Castillo Ochoa, A., Reyna Bello, A., Serrano, A. T., & Fernandez Gomez, R. (2018). Cloning, expression and immunological evaluation of the Omp31 protein of Brucella melitensis and evaluation of its possible use for the diagnosis in bovine brucellosis. Revista De Investigaciones Veterinarias Del Perú, 29(3), 996-1008. https://doi.org/10.15381/rivep.v29i3.14008