el-Zaatari F A, Naser S A, Engstrand L, Hachem C Y, Graham D Y
Veterans Affairs Medical Center, Department of Medicine, Houston, Texas 77030.
Curr Microbiol. 1994 Sep;29(3):177-84. doi: 10.1007/BF01570760.
Mycobacterium paratuberculosis is the causative agent of Johne's disease, a chronic enteritis in ruminants, and it has also been isolated and identified from patients with Crohn's disease, an inflammatory bowel disease. The control of Johne's disease has been hampered by the lack of a reliable diagnostic test because of the large degree of antigenic cross-reactivity between mycobacterial and non-mycobacterial species. To help identify specific antigen(s) or epitope(s), an M. paratuberculosis expression library was screened with antibodies and DNA probes. In total, 54 clones were randomly picked, purified, and characterized by DNA probes and monoclonal antibodies with known specificity to individual mycobacterial antigens. Four clones carrying the heat shock protein 65K-, two representing the secreted protein 32K-, three representing the 21K-, and 20 clones representing the specific insertion element of M. paratuberculosis (IS900)-encoding genes and their gene products were identified and characterized. Well-defined recombinant antigens and/or epitopes representing M. paratuberculosis may facilitate the development of specific diagnostic tests and the investigation of their role in these chronic diseases.
副结核分枝杆菌是反刍动物慢性肠炎——约内氏病的病原体，并且也已从炎症性肠病——克罗恩病患者中分离和鉴定出来。由于分枝杆菌与非分枝杆菌物种之间存在很大程度的抗原交叉反应性，缺乏可靠的诊断测试阻碍了约内氏病的防控。为了帮助鉴定特异性抗原或表位，用抗体和DNA探针筛选了副结核分枝杆菌表达文库。总共随机挑选了54个克隆，进行纯化，并通过对单个分枝杆菌抗原具有已知特异性的DNA探针和单克隆抗体进行表征。鉴定并表征了四个携带65K热休克蛋白的克隆、两个代表32K分泌蛋白的克隆、三个代表21K蛋白的克隆，以及20个代表副结核分枝杆菌特异性插入元件（IS900）编码基因及其基因产物的克隆。明确界定的代表副结核分枝杆菌的重组抗原和/或表位可能有助于开发特异性诊断测试，并有助于研究它们在这些慢性疾病中的作用。
