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Int. J. Mol. Sci. 2016, 17(5), 659; doi:10.3390/ijms17050659

Comprehensive Identification of Immunodominant Proteins of Brucella abortus and Brucella melitensis Using Antibodies in the Sera from Naturally Infected Hosts

1
Institute of Animal Hygiene and Environmental Health, Centre for Infectious Medicine, Freie Universität Berlin, Robert-von-Ostertag-Str. 7-13, Berlin 14163, Germany
2
Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Institute of Bacterial Infections and Zoonoses, Naumburger Str. 96a, Jena 07743, Germany
3
Faculty of Veterinary Medicine, Benha University, Moshtohor, Toukh 13736, Egypt
4
Institute of Chemistry and Biochemistry, Freie Universität Berlin, Thielallee 63, Berlin 14195, Germany
*
Author to whom correspondence should be addressed.
Academic Editor: Patrick C.Y. Woo
Received: 11 March 2016 / Revised: 15 April 2016 / Accepted: 22 April 2016 / Published: 30 April 2016
(This article belongs to the Special Issue Host-Microbe Interaction)
View Full-Text   |   Download PDF [2072 KB, uploaded 30 April 2016]   |  

Abstract

Brucellosis is a debilitating zoonotic disease that affects humans and animals. The diagnosis of brucellosis is challenging, as accurate species level identification is not possible with any of the currently available serology-based diagnostic methods. The present study aimed at identifying Brucella (B.) species-specific proteins from the closely related species B. abortus and B. melitensis using sera collected from naturally infected host species. Unlike earlier reported investigations with either laboratory-grown species or vaccine strains, in the present study, field strains were utilized for analysis. The label-free quantitative proteomic analysis of the naturally isolated strains of these two closely related species revealed 402 differentially expressed proteins, among which 63 and 103 proteins were found exclusively in the whole cell extracts of B. abortus and B. melitensis field strains, respectively. The sera from four different naturally infected host species, i.e., cattle, buffalo, sheep, and goat were applied to identify the immune-binding protein spots present in the whole protein extracts from the isolated B. abortus and B. melitensis field strains and resolved on two-dimensional gel electrophoresis. Comprehensive analysis revealed that 25 proteins of B. abortus and 20 proteins of B. melitensis were distinctly immunoreactive. Dihydrodipicolinate synthase, glyceraldehyde-3-phosphate dehydrogenase and lactate/malate dehydrogenase from B. abortus, amino acid ABC transporter substrate-binding protein from B. melitensis and fumarylacetoacetate hydrolase from both species were reactive with the sera of all the tested naturally infected host species. The identified proteins could be used for the design of serological assays capable of detecting pan-Brucella, B. abortus- and B. melitensis-specific antibodies. View Full-Text
Keywords: Brucella; host specificity; mass spectrometry; Liquid chromatography–mass spectrometry (LC-MS); two dimensional electrophoresis; 2D-PAGE; Matrix-assisted laser desorption/ionization-Tine of Flight-Mass Spectrometry MALDI-TOF MS; proteomics; Western blot Brucella; host specificity; mass spectrometry; Liquid chromatography–mass spectrometry (LC-MS); two dimensional electrophoresis; 2D-PAGE; Matrix-assisted laser desorption/ionization-Tine of Flight-Mass Spectrometry MALDI-TOF MS; proteomics; Western blot
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Wareth, G.; Eravci, M.; Weise, C.; Roesler, U.; Melzer, F.; Sprague, L.D.; Neubauer, H.; Murugaiyan, J. Comprehensive Identification of Immunodominant Proteins of Brucella abortus and Brucella melitensis Using Antibodies in the Sera from Naturally Infected Hosts. Int. J. Mol. Sci. 2016, 17, 659.

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