title: An evaluation of ELISA using recombinant Brucella abortus bacterioferritin (Bfr) for bovine brucellosis.
abstract: To date, detection of antibodies against the lipopolysaccharide portion is the backbone of most serodiagnostic methods for brucellosis screening. However this pose a risk for false positive reactions related to other pathogens especially that of Yersinia enterocolitica O:9 which has the most prominent cross reactivity with Brucella spp. In this study, cloning and expression of Brucella abortus bacterioferritin (Bfr) was accomplished by PCR amplification into an expression vector system, and purification of a recombinant B. abortus Bfr (rBfr). The immunogenicity of rBfr was confirmed by Western blot with Brucella-positive bovine serum. To determine whether rBfr has a potential benefit for use in the serodiagnosis of bovine brucellosis, rBfr-based ELISA was performed. Interestingly, rBfr was able to detect anti-Brucella antibodies in positive sera in a dependent manner of TAT values but did not show an immunoreaction with negative samples. Particularly, average OD492 values at the lowest, medium and highest TAT titer levels were 1.4, 2.2 and 2.6-fold increase compared with the cutoff value, respectively. The accuracy, specificity and sensitivity of rBfr showed 89.09%, 93.6% and 85.33%, respectively. These findings suggest that rBfr might be a good candidate for serological diagnosis development of bovine brucellosis.
authors.affiliation: Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, Republic of Korea.
authors.forename: Huynh Tan
authors.initials: HT
authors.lastname: Hop
authors.affiliation: Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, Republic of Korea.
authors.forename: Lauren Togonon
authors.initials: LT
authors.lastname: Arayan
authors.affiliation: Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, Republic of Korea.
authors.forename: Hannah Leah
authors.initials: HL
authors.lastname: Simborio
authors.affiliation: Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, Republic of Korea.
authors.forename: Alisha Wehdnesday Bernardo
authors.initials: AW
authors.lastname: Reyes
authors.affiliation: Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, Republic of Korea.
authors.forename: WonGi
authors.initials: W
authors.lastname: Min
authors.affiliation: Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, Republic of Korea.
authors.forename: Hu Jang
authors.initials: HJ
authors.lastname: Lee
authors.affiliation: 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: Institute of Agriculture and Life Science, Gyeongsang National University, Jinju 660-701, Republic of Korea.
authors.forename: Hong Hee
authors.initials: HH
authors.lastname: Chang
authors.affiliation: Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, Republic of Korea; Institute of Agriculture and Life Science, Gyeongsang National University, Jinju 660-701, Republic of Korea. Electronic address: kimsuk@gnu.ac.kr.
authors.forename: Suk
authors.initials: S
authors.lastname: Kim
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood*; D000942:Antigens, Bacterial / Q000276:immunology; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D015153:Blotting, Western; D002003:Brucella abortus / Q000235:genetics / Q000276:immunology*; D002007:Brucellosis, Bovine / Q000175:diagnosis*; D002417:Cattle; D003001:Cloning, Molecular; D003429:Cross Reactions; D003573:Cytochrome b Group / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D005189:False Positive Reactions; D005293:Ferritins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification; D012698:Serologic Tests / Q000662:veterinary*; D015008:Yersinia enterocolitica / Q000276:immunology
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D003573:Cytochrome b Group; D011994:Recombinant Proteins; D005293:Ferritins; C018032:bacterioferritin
keywords: Bfr; Brucella abortus; ELISA; Recombinant protein; Serodiagnosis