소 브루셀라병 진단용 고면역원성 항원의 추출과 검증
Extraction and verification of highly immunologenic antigen for diagnosis of bovine brucellosis
2
조회
0
다운로드
1
인용
0
참고문헌
키워드
초록
Bovine brucellosis, an important zoonosis, is diagnosed with serological tests such as the RBT, TAT using inactivated whole bacterial cells or bacterial lipopolysaccharide (LPS) antigen in Korea. However, a strong cross-reaction between Brucella spp. and Yersinia enterocolitica O9 in these tests has seriously complicated the diagnosis of animal brucellosis because Brucella spp. shares common antigenic determinants with Y. enterocolitica O9 in the smooth LPS region. In this study, Brucella-field strains were isolated from Brucellapositive Hanwoo in Kimhae, Korea and outer membrane protein (omp) which has low cross-reaction with Y. enterocolitica O9 and high immunogenicity was extracted from the field strains Then we compared ELISA using the extract with RBT-TAT. Fifteen field strains were isolated from 47 supra-mammary-lymph nodes, which were collected from 18 farms. Isolation rate was 32%. Brucella-specific antigen was identified by performing SDS-PAGE or Western blotting on extracted omp with at 0.5% n-lauroylsarcosine One hundred and ninety-two serum-samples were used in the experiment: 142 negative and 50 positive samples verified by RBT-TAT. According to ELISA results, 127 samples were negative and 15 appeared positive among 142 negatives by RBT-TAT, while 42 samples were positive and 8 were negative among 50 positives by RBT-TAT. Therefore, it showed 89.4% of specificity and 84% of sensi-tivity. Through the current experiments, we could set up an ELISA based on the omp which has low cross-reaction and high immunogenicity and concluded that the omp could be a good material for accurate diagnosis of bovine brucellosis.
Abstract
Bovine brucellosis, an important zoonosis, is diagnosed with serological tests such as the RBT, TAT using inactivated whole bacterial cells or bacterial lipopolysaccharide (LPS) antigen in Korea. However, a strong cross-reaction between Brucella spp. and Yersinia enterocolitica O9 in these tests has seriously complicated the diagnosis of animal brucellosis because Brucella spp. shares common antigenic determinants with Y. enterocolitica O9 in the smooth LPS region. In this study, Brucella-field strains were isolated from Brucellapositive Hanwoo in Kimhae, Korea and outer membrane protein (omp) which has low cross-reaction with Y. enterocolitica O9 and high immunogenicity was extracted from the field strains Then we compared ELISA using the extract with RBT-TAT. Fifteen field strains were isolated from 47 supra-mammary-lymph nodes, which were collected from 18 farms. Isolation rate was 32%. Brucella-specific antigen was identified by performing SDS-PAGE or Western blotting on extracted omp with at 0.5% n-lauroylsarcosine One hundred and ninety-two serum-samples were used in the experiment: 142 negative and 50 positive samples verified by RBT-TAT. According to ELISA results, 127 samples were negative and 15 appeared positive among 142 negatives by RBT-TAT, while 42 samples were positive and 8 were negative among 50 positives by RBT-TAT. Therefore, it showed 89.4% of specificity and 84% of sensi-tivity. Through the current experiments, we could set up an ELISA based on the omp which has low cross-reaction and high immunogenicity and concluded that the omp could be a good material for accurate diagnosis of bovine brucellosis.
참고문헌 (32)
- 국립수의과학검역원, , 동물 브루셀라병 국제 심포지움, pp.1~163 (2006)
- 김석, "브루셀라 감염증의 발병 기전: 브루셀라 균의 병원성인자", 한국수의공중보건학회지, 31(2), pp.105~114 (2007)
- 김금화, "브루셀라병 검색에 사용되는 여러 가지 혈청학적 진단법의 비교연구", 대한수의학회지, 22(2), pp.149~153 (1982)
- 김병수, "Brucella abotus 119-3균의OMP 항원과 타병원성 균간의 교차반응에 관한 연구", 한국수의공중보건학회지, 22(1), pp.9~15 (1998)
- 농림부, "소 부루셀라병표준방역지침", 3~88 (2006)
- 농림부, "제 2006-34 (2006.06.26)호. 결핵병 및 브루셀라병방역실시요령", ~ (2006)
- 백병걸, "한우에 있어서 불활화 Brucella abortus, Brucella melitensis 그리고 Yersinia enterocolitica에 의한 면역반응 관찰", 한국수의공중보건학회지, 22(2), pp.93~102 (1998)
- 백병걸, "소 브루셀라병 진단을 위한 라텍스 응집반응 시험법", 한국수의공중보건학회지, 23(3), pp.153~163 (1999)
- 손준용, "Zoonosis 브루셀라증에 관한 연구", 국립보건원보, 23, pp.281~295 (1986)
- 윤하정, "환례-대조군 연구를 이용한 국내 소 브루셀라병 발생위험 요인 분석", 한국수의공중보건학회지, 30(1), pp.77~82 (2006)
- 임윤규, "SDS 처리한 브루셀라 항원과 Yersiniaenterocolitica 0:9주의 혈청학적 교차반응 연구", 대한수의학회지, 35(1), pp.43~148 (1995)
- 임현술, "브루셀라증의 개요", 한국역학회지, 27(1), pp.26~36 (2005)
- Bricker BJ, "Evaluation of the Brucella abortus species-specific polymerase chain reaction assay, an improved version of the Brucella AMOS polymerase chain reaction assay for cattle", J Vet Diagn Invest, 15(4), pp.374~378 (2003)
- Corbel MJ, "Differentiation of the serological response to Yersinia enterocolitica and Brucella abortus in cattle", J Hyg December, 68(4), pp.519~530 (1970)
- De Klerk E, "Comparative evaluation of the enzyme-linked immunosorbent assay in the laboratory diagnosis of brucellosis", J Clin Microbiol, 21(3), pp.381~386 (1985)
- Delpino MV, "Occurrence and potential diagnostic applications of serological crossreactivities between Brucella and other alpha-proteobacteria", Clin Diagn Lab Immunol, 11(5), pp.868~873 (2004)
- Dohoo IR, "A comparison of five serological tests for bovine brucellosis", Can J Vet Res, 50(4), pp.485~493 (1986)
- Erdenebaatar J, "Enzyme-linked immunosorbent assay to differentiate the antibody responses of animals infected with Brucella species from those of animals infected with Yersinia enterocolitica O9", Clin Diagn Lab Immunol, 10(4), pp.710~714 (2003)
- Erdenebaatar J, "Epidemiological and serological survey of brucellosis in Mongolia by ELISA using sarcosine extracts", Microbiol Immunol, 48(8), pp.571~577 (2004)
- Ewalt DR, "Validation of the abbreviated Brucella AMOS PCR as a rapid screening method for differentiation of Brucella abortus field strain isolates and the vaccine strains, 19 and RB51", J Clin Microbiol, 38(8), pp.3085~3086 (2000)
- Harris HJ, "Chronic Brucellosis-The Unsatisfactory Status of Current Diagnostic Methods", Am J Public Health Nations Health, 39(7), pp.870~874 (1949)
- Kim JW, "Evaluation of immunochromatographic assay for serodiagnosis of Brucella canis", J Vet Med Sci, 69(11), pp.1103~1107 (2007)
- Mainar-Jaime RC, "Specificity dependence between serological tests for diagnosing bovine brucellosis in Brucella-free farms showing false positive serological reactions due to Yersinia enterocolitica O:9", Can Vet J, 46(10), pp.913~916 (2005)
- McGiven JA, "A new homogeneous assay for high throughput serological diagnosis of brucellosis in ruminants", J Immunol Methods, 337(1), pp.7~15 (2008)
- Mitka S, "Evaluation of different PCR assays for early detection of acute and relapsing brucellosis in humans in comparison with conventional methods", J Clin Microbiol, 45(4), pp.1211~1218 (2007)
- Muma JB, "Evaluation of three serological tests for brucellosis in naturally infected cattle using latent class analysis", Vet Microbiol, 125(1), pp.187~192 (2007)
- Mun˜oz PM, "Efficacy of several serological tests and antigens for diagnosis of bovine brucellosis in the presence of false-positive serological results due to Yersinia enterocolitica O:9", Clin Diagn Lab Immunol, 12(1), pp.141~151 (2005)
- Nielsen K, "Validation of a second generation competitive enzyme immunoassay (CELISA) for the diagnosis of brucellosis in various species of domestic animals", Vet Immunol Immunopathol, 12(3), pp.246~250 (2008)
- OIE, "Manual of diagnostic tests and vaccines for terrestrial animals", pp.409~438 (2004)
- Riezu-Boj JI, "Comparison of lipopolysaccharide and outer membrane protein-lipopolysaccharide extracts in an enzyme-linked immunosorbent assay for the diagnosis of Brucella ovis infection", J Clin Microbiol, 23(5), pp.938~942 (1986)
- Watarai M, "A rapid agglutination assay for canine brucellosis using antigen coated beads.J Vet Med Sci 69(5):477-480", J Vet Med Sci, 69(5), pp.477~480 (2007)
- Wellinghausen N, "Rapid detection of Brucella spp. in blood cultures by fluorescence in situ hybridization", J Clin Microbiol, 44(5), pp.1828~1830 (2006)