Serodiagnosis of tuberculosis using an ELISA with antigen 5 and a hemagglutination assay with glycolipid antigens. Results in patients with newly diagnosed pulmonary tuberculosis ranging in extent of disease from minimal to extensive.

Hemagglutination tests with three glycolipid antigens, A1, B1, and C, and ELISA with antigen 5 were done on serum from Chinese patients with pulmonary tuberculosis and from normal subjects in Hong Kong. Tests with all four antigens were of similar efficiency, giving positive results in 30 to 52% of 88 smear-positive patients, in 16 to 22% of 37 smear-negative, culture-positive patients, in 5 to 13% of 76 culture-negative patients with radiologically active disease, in 5 to 11% of 217 culture-negative patients with inactive disease, and in 1 to 4% of 140 normal subjects. If tests were combined so that an overall positive was scored when all tests were positive, there was worse discrimination between patients and normal subjects; however, as suggested by the poor correlation between the results with pairs of the tests, better discrimination was obtained if an overall positive was scored when any of the tests was positive.

Resistance to rifampicin and isoniazid in strains of Mycobacterium tuberculosis.

Drug susceptibility studies on strains of Mycobacterium tuberculosis isolated from widely different populations of patients and tested by two different techniques indicated that all 55 strains resistant to rifampicin were also resistant to isoniazid, while many strains resistant to isoniazid were found to be susceptible to rifampicin. This observation, which has as yet unknown laboratory and clinical significance, may be particularly useful in management of patients. Further studies are called for to establish this relation.

Hemagglutination tests for tuberculosis with mycobacterial glycolipid antigens. Results in patients with active pulmonary tuberculosis before and during chemotherapy and in healthy tuberculosis contacts.

Hemagglutination tests using three serologically active mycobacterial glycolipids as antigens were carried out on serum specimens from patients with pulmonary tuberculosis and from healthy family contacts of patients with tuberculosis in Singapore. A positive response to any of the three antigens was found in 82.5% of 211 patients with newly diagnosed disease and in 21% of 100 contacts.

Comparison of enzyme-linked immunosorbent assay and hemagglutination test using mycobacterial glycolipids.

Enzyme-linked immunosorbent assay and hemagglutination methods were compared using mycobacterial glycolipids as antigens. Both methods were found to have equivalent specificity and sensitivity in detecting mycobacterial diseases. Both tests had 96% specificity; the sensitivity of the enzyme-linked immunosorbent assay was 86%, and that of the hemagglutination test was 88.

Rapid drug-susceptibility testing of Mycobacterium tuberculosis.

Three methods for rapidly determining the susceptibility of Mycobacterium Tuberculosis isolates to isoniazid, rifampin, ethambutol, streptomycin, and para-aminosalicylic acid were evaluated in a large-scale, blind study. Two of the methods measured evolution of CO2 from radio-labeled substrate (14CO2), and one method measured incorporation of 3H-uracil into ribonucleic acid. Rapid indirect drug-susceptibility test results for nearly 300 isolates were compared with those obtained using a standard modified proportion technique.

ELISA tests for antibodies against mycobacterial glycolipids.

ELISA tests with purified mycobacterial glycolipids and bovine heart cardiolipin are described. The possible clinical use of ELISA tests with mycobacterial glycolipids for the diagnosis of tuberculosis and other mycobacterioses is discussed.

Antitumor activity of mycobacterial glycolipid A1.

Glycolipid A1 isolated from Mycobacterium bovis BCG, when dissolved in olive oil and injected together with Line 10 transplantable hepatoma cells, is able to elicit a host response which results in the abrogation or retardation of tumor growth in syngeneic guinea pigs. Glycolipid A1 does not have adjuvant activity for delayed type hypersensitivity, and antibodies to A1 have not been detected in the sera of guinea pigs during or after the tumor abrogation induced by A1 injection Glycolipid A1 does not share antigenic determinants with Line 10 cell lipid fractions. The possible role of the granuloma response elicited by A1 in controlling tumor growth is discussed.

Automatable radiometric detection of growth of Mycobacterium tuberculosis in selective media.

The formulation of media for selective, automatable, radiometric detection of growth of Mycobacterium tuberculosis in vitro is described. Palmitic-1 acid labeled with carbon-14 and formic-14C acid were compared as substrate sources of [14C]O2 in media deficient in carbohydrate and containing appropriate antimicrobial agents that are not active against tubercle bacilli. A preliminary clinical laboratory study of a medium containing 4 microCi palmitic-1-14C acid per ml showed that this method might provide the basis for practical laboratory use.

Granulomagenic activity of serologically active glycolipids from Mycobacterium bovis BCG.

The granulomagenic properties of serologically active glycolipids A1, B2, B3, and C isolated from Mycobacterium bovis BCG were studied. Glycolipid A1, dissolved in olive and injected intradermally in guinea pigs, was able to elicit a granulomatous response that seemed to be of the nonallergic type. This granulomagenic activity was quite striking since only 2 mug was necessary to elicit the reaction.

Serologically active glycolipid families from Mycobacterium bovis BCG. I. Extraction, purification and immunologic studies.

Glycolipids were extracted from mycobacteria with methanol and chloroform and purified by silicic acid chromatography. These glycolipids were studied for their serologic activity by direct and indirect (Coomb's) passive hemagglutination, and by inhibition methods. Three families of serologically active glycolipids called A, B and C, plus cardiolipin, were isolated.

Serologically active glycolipid families from Mycobacterium bovis BCG. II. Serologic studies on human sera.

Serological tests with serologically active glycolipids from Mycobacterium bovis BCG were investigated for their possible use in the diagonis of mycobacterioses. The results were positive with 95 percent of sera from patients with far advanced and moderately advanced tuberculosis, 80 percent of sera from patients with newly diagnosed tuberculosis and 75 percent of patients with atypical mycobacterial diseases. "False" positive serologic reactions were obtained from 4 percent or less.

Delayed-type hypersensitivity and immunity against aerogenic tuberculosis in guinea pigs.

In experiments described herein, it was observed that guinea pigs with delayed-type hypersensitivity to tuberculoproteins under various experimental conditions, with or without passive transfer of serum from immune donors, manifested no acquired immunity against aerogenically induced tuberculosis. These results are discussed in relation to the previous observations of other investigators and to the general problem of cellular hypersensitivity and immunity against facultative intracellular bacteria.

Continuous dispenser for multiple-well serological plate.

Materials and methods are described for the construction of an inexpensive multiple-well dispensing apparatus for use in serological microtitration.