Serodiagnosis of tuberculosis: detection of mycobacterial antibodies and antigens.

SETTING: 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. OBJECTIVE: 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. DESIGN: 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. RESULTS: 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. CONCLUSION: 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.

[Detection of Mycobacterial antigens in tuberculous pleuritis, empyema and meningitis]. / Opredelenie mikobakterialnykh antigenov pri tuberkulëznykh plevrite, émpieme i meningite.

Mycobacterial antigens were identified by inhibition solid-phase enzyme immunoassay specific antibodies to M.H37Rv labelled with horse radish peroxidase in 20, 27, 24, 47 and 21 patients with tuberculous pleurisy, tuberculous empyema, meningitis, pleurisy and nontuberculous empyema, respectively. Mycobacterial antigens were found more frequently and in greater quantities in pleural and cerebrospinal fluids in tuberculosis than in the above nontuberculous affections. These differences were less pronounced in serum assays.

[Dynamics in bacterial isolation and antigenemia in patients with infiltrative pulmonary tuberculosis]. / Dinamika bakteriovydeleniia i antigenemii u bolnykh infiltrativnym tuberkulëzom lëgkikh.

A hundred patients with infiltrative pulmonary tuberculosis were studied. They were examined for bacterial isolates and mycobacterial antigens in the immune complexes. The isolates were detected in 76%. In most cases, isolation stopped 3 months following chemotherapy and in the absolute majority of cases (except one) 9 months after. Isolation of L-forms remained for a longer period of time, at month 6 of therapy there was an increase in the number of patients who were found to have L-forms of the bacteria. Most (97%) patients with infiltrative tuberculosis displayed mycobacterial antigens circulating in the immune complexes. Antigenemia retained in a considerable (40%) number of patients long (up to 9 months) and in most cases with the unfavourable course of the disease.

[Identification of mycobacteria in sputum by determining antigens in the culture medium]. / Identifikatsiia mikobakterii v mokrote s pomoshchíu opredeleniia antigenov v kulturalnoi zhidkosti.

Antigens to M. tuberculosis in culture fluid of sputum samples were compared to bacterioscopy and inoculation results for 64 new-onset cases of tuberculosis and 60 chronics with bronchitis. The authors used diagnostic preparations of monoclonal and polyclonal antibodies to M. tuberculosis. The use of the above preparations in the enzyme immunoassay reduced the time of M. tuberculosis detection in the patients' sputum and increased the frequency of the bacteria registration against standard methods of bacterioscopy and inoculation.

[Production of purified antigenic drug from Mycobacterium tuberculosis humanus and its use for serodiagnosis of tuberculosis]. / Poluchenie ochishchennogo antigennogo preparata iz mikobakterii tuberkulëza vida Humanus i ego ispolzovanie dlia serodiagnostiki tuberkulëza.

A variety of antigenic preparations from whole bacteria M. tuberculosis humanus (H37Rv) and relevant subcellular components have been tested using indirect ELISA for determination of antituberculous antibodies. Cell membranes of mycobacteria and antigens isolated from KCl-extracts of cell membranes in gel Toyopearl HW 55 F demonstrated the highest activity. Fraction 6 usage raised the reaction sensitivity to 83.3% in specificity 86.7%. As indicated by the ELISA, the highest antibodies content under the above antigen introduction was reached in patients with fibrocavernous tuberculosis, the lowest one in focal tuberculosis.