Korean Journal of Veterinary Research 1997;37(2):349-358.
Studies on Molecular Biological and Immunological Diagnosis of Johne's Disease
Tae-jong Kim1, Yun-sik Kim1, Jae-chun Kim1, Wha-joong Yoon1, Won-chang Lee1, SJ Shin2, YF Chang2
1Department of Veterinary Medicine, College of Animal Husbandary and Animal Resources Research Center, Kon-Kuk University
2College of Veterinary Medicine, Cornell University
분자생물학과 면역학적 방법에 의한 소 요네병 진단의 연구
김태종1, 김윤식1, 김재천1, 윤화중1, 이원창1, 2, 2
1건국대학교 축산대학 수의학과, 동물자원연구센타
2미국 Cornell 대학교 수의과대학
Abstract
Mycobacterium paratuberculosis is the etiologic agent of Johne's disease, a chronic inflammatory bowel syndrome in ruminants. The attempts to control or eradicate the disease were severely hampered by the inadequacies of present diagnostic methods. The first purpose of this study was to detect Johne's disease out of 577 cows in the province of Kyunggi, Chungchong, Gangweon and the second purpose was to compare the results of non-absorbed ELISA, absorbed ELISA, PCR, and conventional culture methods. The third purpose was to increase diagnostic specificity, accuracy and rapidity. When non-absorbed ELISA test was conducted with Mycobacterium paratuberculosis antigen, the prevalence of positive was 10.9%. To increase diagnostic specificity, absorbed ELISA test with Mycobacterium phlei was used. In this test, the positive prevalence was 1.7%. For the specific detection of Mycobacterium paratuberculosis, PCR was applied to bacterial culture obtained from fecal samples of cattle. The DNA sequences derived from IS900 were used to prepare DNA primers for detection and identification of Mycobacterium paratuberculosis by PCR. PCR for M paratuberculosis isolated from fecal cultures amplified specific target DNA. PCR was much more rapid than that obtained by conventional culture technique in diagnosis of Johne's disease.
Key Words: Mycobacterium paratuberculosis, ELISA, PCR, Fecal culture, sequence


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