人型结核杆菌H<sub>37</sub>Ra的抗原决定簇的筛选及其初步鉴定

人型结核杆菌H₃₇Ra的抗原决定簇的筛选及其初步鉴定

摘要: 本文报导一种筛选人型结核杆菌H₃₇Ra的抗原决定簇的Western Blot分析技术。细菌培养滤液经15%SDS-PAGE电泳进行分离,然后电转移到硝酸纤维素薄膜上,应用鼠抗人型结核杆菌单克隆抗体进行免疫反应。通过免疫印染,一条能与单克隆抗体起强阳性反应的蛋白带被识别。应用蛋白质洗脱技术,这个蛋白质片段被提纯。经过SDS-PAGE电泳分析,这个含有抗原决定簇的蛋白质片段分子量为17500道尔顿,斑点免疫试验证实该片段具有生物学活性。本文提示,Western Blot分析技术是选择抗原决定簇的有用方法,为结核病的诊断提供了新的途径。

Abstract: A new procedure for the selection of antigenic determinant of Mycobacterium tuberculosis H₃₇Ra is described. The proteins were purified from culture filtrates by polyacrylamide gel electrophoresis（PAGE）, transferred onto nitrocellulose membrane, immunostained with monoclonal antibodies（McAbs） against Mycobacterium tuberculosis. One McAbs-positive protein antigen was obtained by using the Western Blot analysis. The molecular weight of the determinant was 17500 Dalton by SDS-PAGE analysis. The results suggest that the Western Blot analysis may be a useful tool for the diagnosis of the tuberculosis disease.