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Clinical and Diagnostic Laboratory Immunology, July 2003, p. 647-651, Vol. 10, No. 4
1071-412X/03/$08.00+0     DOI: 10.1128/CDLI.10.4.647-651.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Epitope Mapping of the Brucella melitensis BP26 Immunogenic Protein: Usefulness for Diagnosis of Sheep Brucellosis

Patricia Seco-Mediavilla,1 Jean-Michel Verger,2 Maggy Grayon,2 Axel Cloeckaert,3 Clara M. Marín,4 Michel S. Zygmunt,2 Luis Fernández-Lago,1 and Nieves Vizcaíno1*

Departamento de Microbiología y Genética, Universidad de Salamanca, 37007 Salamanca,1 Unidad de Sanidad Animal, Servicio de Investigación Agroalimentaria, Diputación General de Aragón, 50080 Zaragoza, Spain ,4 Station de Pathologie Infectieuse et Immunologie,2 Unité BioAgresseurs, Santé et Environnement, Institut National de la Recherche Agronomique, 37380 Nouzilly, France3

Received 25 October 2002/ Returned for modification 18 December 2002/ Accepted 21 January 2003

Sequencing of bp26, the gene encoding the Brucella sp. immunogenic BP26 periplasmic protein, was performed in the reference strains of Brucella abortus, B. suis, and B. ovis. The three bp26 sequences were almost identical to that published for B. melitensis 16M bp26, and only minor nucleotide substitutions, without modifying the amino acid sequence, were observed between species. The bp26 genes of the seven B. abortus biovar reference strains and B. abortus S19 and RB51 vaccine strains were also sequenced. Again, only minor differences were found. Surprisingly, the bp26 nucleotide sequence for B. abortus S19 was almost identical to that found for B. melitensis 16M and differed from the sequence described previously by others (O. L. Rossetti, A. I. Arese, M. L. Boschiroli, and S. L. Cravero, J. Clin. Microbiol. 34:165-169, 1996) for the same B. abortus strain. The epitope mapping of BP26, performed by using a panel of monoclonal antibodies and recombinant DNA techniques, allowed the identification of an immunodominant region of the protein interesting for the diagnosis of B. melitensis and B. ovis infection in sheep. A recombinant fusion protein containing this region of BP26 reacted indeed, in Western blotting, as the entire recombinant BP26 against sera from B. melitensis- or B. ovis-infected sheep while it avoided false-positive reactions observed with sera from Brucella-free sheep when using the entire recombinant BP26. Thus, use of this recombinant fusion protein instead the entire recombinant BP26 could improve the specific serological diagnosis of B. melitensis or B. ovis infection in sheep.

* Corresponding author. Mailing address: Departamento de Microbiología y Genética, Edificio Departamental, Universidad de Salamanca, Avda. Campo Charro s/n, 37007 Salamanca, Spain. Phone: 34-923-294532. Fax: 34-923-224876. E-mail: vizcaino{at}usal.es.

Clinical and Diagnostic Laboratory Immunology, July 2003, p. 647-651, Vol. 10, No. 4
1071-412X/03/$08.00+0     DOI: 10.1128/CDLI.10.4.647-651.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Yang, X., Hudson, M., Walters, N., Bargatze, R. F., Pascual, D. W. (2005). Selection of Protective Epitopes for Brucella melitensis by DNA Vaccination. Infect. Immun. 73: 7297-7303 [Abstract] [Full Text]  


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