| Development of PCR assay for the detection of Brucella spp in bovine semen |
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Suck-chan Jung1, Byeong-yeal Jung1, Seong-ryong Woo1, Dong-hee Cho1, Jong-yeom Kim1, Woo-taek Kim2, Jung-mi Lee3, Yong-ho Park3, Byeong-kirl Baek4 |
1Department of Bacteriology and Immunology, National Veterinary Research Institute
2Department of Animal Health, Cheju Livestock Promotion Institute
3College of Veterinary Medicine, Seoul National University
4College of Veterinary Medicine, Chonbuk National University |
| 종모우 정액중 Brucella균 신속 검출을 위한 PCR기법 개발 |
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정석찬1, 정병열1, 우승룡1, 조동희1, 김종염1, 김우택2, 이정미3, 박용호3, 백병걸4 |
1수의과학연구소
2제주도 축산진흥원
3서울대학교 수의과대학
4전북대학교 수의과대학 |
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| Abstract |
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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. |
| Key Words:
Brucella abortus, PCR, bull semen |
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