Korean Journal of Veterinary Research (대한수의학회지)
- Volume 38 Issue 2
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- Pages.345-352
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- 1998
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- 2466-1384(pISSN)
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- 2466-1392(eISSN)
Development of PCR assay for the detection of Brucella spp in bovine semen
종모우 정액중 Brucella균 신속 검출을 위한 PCR기법 개발
- Jung, Suck-chan (Department of Bacteriology and Immunology, National Veterinary Research Institute) ;
- Jung, Byeong-yeal (Department of Bacteriology and Immunology, National Veterinary Research Institute) ;
- Woo, Seong-ryong (Department of Bacteriology and Immunology, National Veterinary Research Institute) ;
- Cho, Dong-hee (Department of Bacteriology and Immunology, National Veterinary Research Institute) ;
- Kim, Jong-yeom (Department of Bacteriology and Immunology, National Veterinary Research Institute) ;
- Kim, Woo-taek (Department of Animal Health, Cheju Livestock Promotion Institute) ;
- Lee, Jung-mi (College of Veterinary Medicine, Seoul National University) ;
- Park, Yong-ho (College of Veterinary Medicine, Seoul National University) ;
- Baek, Byeong-kirl (College of Veterinary Medicine, Chonbuk National University)
- 정석찬 (수의과학연구소) ;
- 정병열 (수의과학연구소) ;
- 우승룡 (수의과학연구소) ;
- 조동희 (수의과학연구소) ;
- 김종염 (수의과학연구소) ;
- 김우택 (제주도 축산진흥원) ;
- 이정미 (서울대학교 수의과대학) ;
- 박용호 (서울대학교 수의과대학) ;
- 백병걸 (전북대학교 수의과대학)
- Received : 1998.03.17
- Published : 1998.06.25
Abstract
The diagnosis of brucellosis is currently based on serological and microbiological tests. However, the microbiological isolation and identification have several disadvantages such as time-consuming and laborious, and the serological methods have been reported to cross-react with antigens other than those from Brucella spp. To develop a sensitive and rapid diagnostic method for detection of Brucella species, the genus-specific primers were designed and synthesized from the sequence of gene encoding a 31kDa cell surface protein(BCSP) and a 36kDa outer membrane protein(OMPB) of B abortus. The amplified 711bp and 982bp DNA fragments were only visible in each species of Brucella by PCR method using the BCSP and OMPB primers, respectively. However, PCR product was not obtained with DNA from other Gram-negative bacteria. As little as 1pg of the B abortus genomic DNA could be detected by this PCR method. Using the PCR technique, semen samples from 185 bulls of Brucella-seronegative herds in Cheju island were examined for comparison of this PCR method with conventional methods in 1995. The semen samples from 5 bulls were positive by culture method and PCR, and one was positive and 5 were suspect by semen plasma agglutination test. However, the semen samples obtained from 177 bulls were negative by semen plasma agglutination, culture and PCR methods in 1996. The results of comparison tests suggested that PCR was a better test than agglutination test against semen of bulls. This study indicated that the PCR technique was a valuable for the diagnosis of bovine brucellosis, particulary in bull semens.