Research Article
Volume 6 Issue 5 - 2021
The Omp31r Recombinant Protein of Brucella melitensis and Immunological Evaluation for its Possible Use for the Diagnosis in Bovine Brucellosis
Annie Castillo Ochoa1, Rodolfo Fernández-Gomez2, Juan Carlos Freites3, Ana Teresa Serrano4 and Jose-David Rosales3*
1Simón Bolívar University, Caracas, Venezuela
2Biophysics Solutions CA, Caracas, Venezuela
3Centre of Agriculture and Alimentarium Security, Institute of Advanced Studies Foundation (IDEA), Caracas, Venezuela
4Faculty of Veterinary Sciences, Central University of Venezuela, Maracay, Venezuela
*Corresponding Author: Jose-David Rosales, Centre of Agriculture and Alimentarium Security, Institute of Advanced Studies Foundation (IDEA), Caracas, Venezuela.
Received: February 08, 2021; Published: April 30, 2021




Abstract

In humans, Brucellosis is a potentially mortal disease, caused by intracellular Gram-negative bacteria, which belongs to the Brucella group and is worldwide distributed. In humans, the infection is mediated through the consumption of milk or fresh cheese no pasteurized and raw meat. Therefore, sanitary control in farm animals is greatly important, thus the serologic diagnosis is key to detect antibodies against Brucella´s lipopolysaccharide, by ELISA tests or agglutination tests. The aim of the work is testing a recombinant protein, to use it as a potential protein for serological diagnosis of brucellosis in bovine. Bioinformatics analysis results in the OMP31 gene identification from Brucella melitensis M15, forward cloned from genomic DNA. The OMP31 protein was expressed in a BL21 (DE3) system, using pET28a vector, and isolated by polyhistidine-NI affinity chromatography. Polyclonal antibodies against OMP31 were produced in rabbit and a western blot against Brucella abortus RB51, Brucella melitensis M15 and Brucella abortus 1119 were performed. The antibodies produced did recognize Brucella abortus RB51 and Brucella melitensis M15, but didn´t recognize the vaccine strain Brucella abortus 1119. A standardized ELISAi was performed, using healthy bovine´s serums and Brucella abortus positive bovine´s serums. The specificity and sensibility were estimated contrasting an ELISAi using OMP31r obtaining 90.6% and 77.17% and Brucella abortus 1119, 100%. The recognition of OMP31r by antibodies from serums of B. abortus infected bovines is quite promising, opening the possibility to produce both, rapids test kit and massive diagnosis to enable the control of bovine brucellosis.

 

Keywords: Brucella sp; Omp31r; Brucellosis; Recombinant Protein; Indirect ELISA

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Citation: Jose-David Rosales., et al. “The Omp31r Recombinant Protein of Brucella melitensis and Immunological Evaluation for its Possible Use for the Diagnosis in Bovine Brucellosis”. EC Veterinary Science 6.5 (2021): 06-16.

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