Characterization of an immuno-dominant antigen in Brucella ovis and evaluation of its use in an enzyme-linked immunosorbent assay

Veterinary Microbiology
R KittelbergerA Cloeckaert

Abstract

A panel of 45 Brucella ovis serologically positive sera were tested in immunoblots against B. ovis outer membrane proteins Omp31 and Omp25, purified by preparative SDS-gel electrophoresis. Forty-three sera reacted with Omp31, while only 11 reacted with Omp25, suggesting that Omp31 is identical to the previously reported immuno-dominant 29-kDa protein. Attempts to purify Omp31 on a larger scale by using procedures such as ion exchange-, reversed phase-, affinity- and gel filtration chromatography suggested that the outer membrane proteins were aggregated with rough lipopolysaccharide. Only denaturing SDS-gel filtration chromatography was able to separate proteins of about 29 kDa from rough lipopolysaccharide but did not separate Omp31 from Omp25 in B. ovis preparations. When used in an enzyme-linked immunosorbent assay, this 29-kDa protein preparation was less sensitive and less specific than the routinely used heat-extracted B. ovis antigen. A readily available recombinant E. coli, expressing the gene for Omp31 from Brucella melitensis 16 M, was used to extract and enrich recombinant Omp31 by a temperature-dependent Triton X-114-based technique. When this material was used in immunoblots with the 45 sera from B. ovis-infected s...Continue Reading

References

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Apr 1, 1995·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·R KittelbergerJ de Bruyn
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Nov 15, 1996·FEMS Microbiology Letters·A CloeckaertN Vizcaíno

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Citations

Nov 5, 2002·Veterinary Microbiology·Axel CloeckaertPhilip H Elzer
Jan 13, 2004·Clinical and Diagnostic Laboratory Immunology·Juliana CassataroPablo C Baldi
Apr 20, 2007·Research in Veterinary Science·Stella Maria Barrouin-MeloSongelí Menezes Freire
Jul 26, 2015·Veterinary Microbiology·P G Gouletsou, G C Fthenakis
Sep 26, 2002·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·Silvia M EsteinRaúl A Bowden
Oct 9, 2018·Frontiers in Microbiology·Rebeca Singh Sidhu-MuñozNieves Vizcaíno

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