Khomenko A G, Bayensky A V, Chernousova L N, Kulikovskaya N V, Demianenko N V, Litvinov V I
Central Research Institute for Tuberculosis, Russian Academy of Medical Science, Moscow, Russia.
Tuber Lung Dis. 1996 Dec;77(6):510-5. doi: 10.1016/s0962-8479(96)90048-9.
The diagnosis of tuberculosis is based primarily on identification of mycobacteria and on clinical evidence. Recently, serological studies have been widely used experimentally as a diagnostic approach.
The aim of our study was to optimize serodiagnosis of tuberculosis by detecting mycobacterial antigens and antibodies in sera from patients with lung tuberculosis, non-related diseases and healthy controls.
Mycobacterium tuberculosis H37Rv was disintegrated by pressure. Cell walls were extracted with 3 M KCL and were subjected to gel filtration in Toyopearl gel. Immune sera were prepared by immunization of rabbits with cell wall material. Anti H37Rv antibodies were purified by affinity chromatography. The reagents obtained were used to detect serum antibodies and antigens (following immune complex dissociation) using ELISA.
Using fraction 6 of cell wall extract, antibodies were detected in 72.2% of TB patients; there were no positive reactions in control subjects. By use of affinity-purified antibodies, antigens were detected in 77.1% of TB patients, 10% of patients with unrelated diseases and 6.7% of healthy controls.
Effective serodiagnosis of tuberculosis can be achieved only by combining detection of both circulating antibodies and antigens using highly specific purified reagents and immune complex-dissociated sera.
结核病的诊断主要基于分枝杆菌的鉴定和临床证据。最近,血清学研究已被广泛用作实验性诊断方法。
我们研究的目的是通过检测肺结核患者、非相关疾病患者和健康对照者血清中的分枝杆菌抗原和抗体,优化结核病的血清学诊断。
结核分枝杆菌H37Rv经压力破碎。用3M KCl提取细胞壁,并在Toyopearl凝胶中进行凝胶过滤。用细胞壁材料免疫兔子制备免疫血清。通过亲和层析纯化抗H37Rv抗体。使用ELISA检测血清抗体和抗原(免疫复合物解离后)。
使用细胞壁提取物的第6部分,72.2%的结核病患者检测到抗体;对照组无阳性反应。使用亲和纯化抗体,77.1%的结核病患者、10%的非相关疾病患者和6.7%的健康对照者检测到抗原。
只有使用高度特异性的纯化试剂和免疫复合物解离血清同时检测循环抗体和抗原,才能实现有效的结核病血清学诊断。