title: Immunoproteomics of Brucella abortus RB51 as candidate antigens in serological diagnosis of brucellosis.
abstract: The current brucellosis serodiagnostic assays are chiefly based on detecting anti-LPS (lipopolysaccharide) antibodies. However, cross-reaction with some gram-negative bacteria can occasionally induce due to similar O-polysaccharide (OPS) structure. Therefore, the aim of the present study was to identify new candidate antigens from Brucella abortus RB51, a mutant strain lacking the LPS portion, which might be valuable in brucellosis diagnosis. To detect potential antigens, immobilized pH gradients (IPG) strips with three ranges (pH 3-5.6, 4-7 and 6-11) were applied. After separating the insoluble proteins of B. abortus RB51 using two-dimensional electrophoresis (2-DE), their immunogenicity was evaluated by western blotting using four types of antisera - B. abortus, Yersinia enterocolitica O:9 and Escherichia coli O157:H7-positive, and B. abortus-negative bovine sera. Among the several immunogenic spots, the spots showing specific reactivity with only the B. abortus-positive antisera, were considered as candidate antigens. Overall, eleven immuno-reactive proteins were identified, as follows: Cu/Zn superoxide dismutase, histidinol dehydrogenase, chaperonin DnaK, chaperonin GroES, beta-ketoadipyl CoA thiolase, two-component response regulator, the cell-division protein FtsZ, aldehyde dehydrogenase, 50s ribosomal protein L10 and invasion protein B. These selected highly immunogenic protein spots might be useful as alternative antigens for brucellosis and helpful in reducing the cross-reactivity.
authors.affiliation: OIE Reference Laboratory for Brucellosis, Bacteriology Division, Animal and Plant Quarantine Agency, Anyang, Gyeonggi do 430-757, Republic of Korea.
authors.forename: Ji-Yeon
authors.initials: JY
authors.lastname: Kim
authors.affiliation: OIE Reference Laboratory for Brucellosis, Bacteriology Division, Animal and Plant Quarantine Agency, Anyang, Gyeonggi do 430-757, Republic of Korea.
authors.forename: So-Ra
authors.initials: SR
authors.lastname: Sung
authors.affiliation: OIE Reference Laboratory for Brucellosis, Bacteriology Division, Animal and Plant Quarantine Agency, Anyang, Gyeonggi do 430-757, Republic of Korea.
authors.forename: Kichan
authors.initials: K
authors.lastname: Lee
authors.affiliation: OIE Reference Laboratory for Brucellosis, Bacteriology Division, Animal and Plant Quarantine Agency, Anyang, Gyeonggi do 430-757, Republic of Korea.
authors.forename: Hyang-Keun
authors.initials: HK
authors.lastname: Lee
authors.affiliation: OIE Reference Laboratory for Brucellosis, Bacteriology Division, Animal and Plant Quarantine Agency, Anyang, Gyeonggi do 430-757, Republic of Korea.
authors.forename: Sung-Il
authors.initials: SI
authors.lastname: Kang
authors.affiliation: OIE Reference Laboratory for Brucellosis, Bacteriology Division, Animal and Plant Quarantine Agency, Anyang, Gyeonggi do 430-757, Republic of Korea.
authors.forename: Jin Ju
authors.initials: JJ
authors.lastname: Lee
authors.affiliation: OIE Reference Laboratory for Brucellosis, Bacteriology Division, Animal and Plant Quarantine Agency, Anyang, Gyeonggi do 430-757, Republic of Korea.
authors.forename: Suk Chan
authors.initials: SC
authors.lastname: Jung
authors.affiliation: OIE Reference Laboratory for Brucellosis, Bacteriology Division, Animal and Plant Quarantine Agency, Anyang, Gyeonggi do 430-757, Republic of Korea.
authors.forename: Yong Ho
authors.initials: YH
authors.lastname: Park
authors.affiliation: OIE Reference Laboratory for Brucellosis, Bacteriology Division, Animal and Plant Quarantine Agency, Anyang, Gyeonggi do 430-757, Republic of Korea. Electronic address: herm@qia.go.kr.
authors.forename: Moon
authors.initials: M
authors.lastname: Her
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D000918:Antibody Specificity; D000942:Antigens, Bacterial / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000276:immunology* / Q000302:isolation & purification; D015153:Blotting, Western; D002003:Brucella abortus / Q000235:genetics / Q000276:immunology*; D002007:Brucellosis, Bovine / Q000175:diagnosis* / Q000276:immunology; D002417:Cattle; D003429:Cross Reactions; D015180:Electrophoresis, Gel, Two-Dimensional; D040901:Proteomics; D012698:Serologic Tests; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001426:Bacterial Proteins
keywords: 2-DE; Brucella abortus RB51; Immunogenic protein; Western blotting