sodium-dodecyl-sulfate and Tuberculosis--Pulmonary

To evaluate specimen processing with sodium dodecyl (lauryl) sulfate (SDS) on the detection of Mycobacterium tuberculosis by PCR-reversed dot hybridization.. Totally 266 specimens were collected from sputum, pleural fluid, ascites, urine or blood of patients with active tuberculosis and were treated with SDS or routine method. All specimens handled were tested for detecting M. tuberculosis by PCR-reversed dot hybridization.. The detection rate of M. tuberculosis from specimens prepared by SDS method was 65.0% (173/266) as compared with 51.5% (136/168) by routine method. Results from two methods were considered statistically significant (chi(2) = 35.03, P < 0.001).. The SDS processing method may be used to detect M. tuberculosis from clinical specimens by PCR and enhances the detection rate of M. tuberculosis.

Topics: Detergents; Humans; Mycobacterium tuberculosis; Peritonitis, Tuberculous; Polymerase Chain Reaction; Sodium Dodecyl Sulfate; Tuberculosis; Tuberculosis, Meningeal; Tuberculosis, Pleural; Tuberculosis, Pulmonary; Tuberculosis, Renal

The concentrations of sodium dodecyl (lauryl) sulfate (SDS) found in the sediments of the respiratory samples pretreated with SDS-NaOH varied between 3.36 and 12.42 mg/ml. These concentrations of SDS were higher than the level considered critical (> or = 0.16 mg/ml) to obtain negative results with the Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test. Extensive washing, after neutralization of sediment, is sufficient to remove all traces of detergent which interfere with the enzymes used by the Mycobacterium Tuberculosis Direct Test assay.

Topics: Bronchoalveolar Lavage Fluid; Detergents; Humans; Mycobacterium tuberculosis; Nucleic Acid Amplification Techniques; RNA, Bacterial; RNA, Ribosomal; Sodium Dodecyl Sulfate; Sodium Hydroxide; Sputum; Tuberculosis, Pulmonary

The perchloric-acid-soluble Bacille bilié de Calmette-Guérin (BCG) antigen (BCG-P) was investigated for the diagnosis of tuberculosis. Monoclonal antibodies against BCG-P were produced. After SDS-PAGE, silver staining and immunoblotting were done. Pathological specimens of pulmonary tuberculosis, BCG organism itself and atypical mycobacterium were stained using the indirect immunofluorescent technique with the strongest one among these monoclonal antibodies. Intradermal skin test and ELISA were also done with BCG-P. BCG-P was composed of a major 65-kD protein and a minor 61-kD protein. Fourteen monoclonal antibodies were produced which were all IgM. These antibodies reacted with the 65-kD protein which also showed cross-reactivity with Mycobacterium tuberculosis in pathologic tissues and atypical mycobacterium. BCG-P did not react with the sera of patients with active pulmonary tuberculosis. However, intradermal skin test with BCG-P showed positive reactions in the patients with a positive purified protein derivative of tuberculin skin test. This antigen could be a good antigen to evaluate the success of BCG vaccination.

Topics: Antibodies, Monoclonal; Antibody Formation; Antigens, Bacterial; BCG Vaccine; Child; Child, Preschool; Electrophoresis, Polyacrylamide Gel; Fluorescent Antibody Technique; Humans; Hypersensitivity, Delayed; Mycobacterium bovis; Perchlorates; Sodium Dodecyl Sulfate; Solubility; Tuberculin Test; Tuberculosis, Pulmonary; Vaccination

The antigens from immune complexes of sera from patients with mycobacterial diseases were released by sodium dodecyl sulfate. The antigenic activity of the released proteins was tested by agar gel diffusion and immunoelectrophoresis. This simple method provided direct evidence for the presence of mycobacterial antigens in the immune complexes of sera from patients with leprosy and tuberculosis.

Topics: Antigen-Antibody Complex; Antigens, Bacterial; Humans; Immunodiffusion; Immunoelectrophoresis; Leprosy; Mycobacterium leprae; Mycobacterium tuberculosis; Sodium Dodecyl Sulfate; Tuberculosis, Pulmonary

Topics: Culture Media; Humans; Microbiological Techniques; Sodium Dodecyl Sulfate; Sputum; Tuberculosis, Pulmonary