Evaluation of recombinant 28 kDa outer membrane protein of Brucella abortus for the clinical diagnosis of bovine brucellosis in Korea.
Brucella spp. are Gram-negative, facultative, intracellular coccobacilli that are pathogenic to a variety of mammals, including ruminants and humans. The conventional serological test for diagnosing brucellosis in cattle in Korea is the standard tube agglutination test. However, agglutination tests sometimes give false-positive results due to cross-reactions with other pathogens. The outer membrane proteins of Brucella species have been extensively studied for their immunogenicity and serodiagnostic applications. However, an application of B. abortus OMPs for serodiagnosis has not been successfully established. In this study, cloning and expression of B. abortus Omp28, a group 3 antigen, were accomplished by PCR amplification cloning into a pMAL expression system, and purification of a recombinant Omp28 (rOmp28). The immunogenicity of rOmp28 was confirmed by Western blot analysis with Brucella-positive bovine serum. To determine whether rOmp2 has a potential benefit for use in the serodiagnosis of bovine brucellosis, rOmp28-based ELISA and latex bead agglutination test were performed. B. abortus positive (n=122) or negative (n=88) from TAT were positive (118/122, 96.7%) or negative (84/88, 95.4%) in ELISA and were positive (94/122, 77%) or negative (71/88, 81.7%) in that the latex bead agglutination test, respectively. The sensitivity, specificity and accuracy were 96.7, 95.4, 96.2% in ELISA and 77, 80.6, 78.5% in latex bead agglutination test, respectively. These findings suggest that the rOmp28 of B. abortus might be a good candidate for developing serological diagnostic tools for bovine brucellosis.
College of Veterinary Medicine, Gyeongsang National University, Jinju 660-701, Republic of Korea.
Jeong Ju
JJ
Lim
Dong Hyeok
DH
Kim
Jin Ju
JJ
Lee
Dae Geun
DG
Kim
Wongi
W
Min
Hu Jang
HJ
Lee
Man Hee
MH
Rhee
Hong Hee
HH
Chang
Suk
S
Kim
D000818:Animals; D015153:Blotting, Western / Q000662:veterinary; D002003:Brucella abortus / Q000235:genetics*; D002007:Brucellosis, Bovine / Q000175:diagnosis*; D002417:Cattle; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D007723:Korea; D007841:Latex Fixation Tests / Q000662:veterinary; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D012680:Sensitivity and Specificity
C106040:BP26 protein, Brucella; D017931:DNA Primers; D008565:Membrane Proteins; D011994:Recombinant Proteins