Binding assay for the detection of mycobacteria
AssigneeMontefiore Medical Center
InventorEugene W. Straus
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Abstract
A method for detecting the presence of Mycobacteria in a fluid or tissue which comprises mixing the fluid or tissue containing a secretory product of Mycobacteria with a complex of a tracer-containing molecule and a binding macromolecule having reversible binding affinity for the tracer-containing molecule and detecting the tracer-containing molecule, wherein the tracer-containing molecule is a charcoal-adsorbable protein from Mycobacterium tuberculosis which has a molecular weight of 20,000-30,000 and which is immunochemically stable from 4.degree.0 C. to 250.degree. C. and has a pH range from 3.0 to 9.0. The method is particularly applicable to the detection of infectious tuberculosis in humans and determining the antibiotic sensitivity of infecting Mycobacteria.
Claims36
A heat stable charcoal-adsorbable labeled protein obtained from the growth media of M. tuberculosis, said protein having a molecular weight of between 20,000 and 30,000, immunochemical stability over a temperature range of 4.degree. C.-250.degree. C., a pH range of 3.0-9.0 and exhibiting comigration towards the anode in the presence of bovine serum albumin and borate buffer at a pH of 8.6 with an applied voltage of 80 volts on starch gel electrophoresis for 12 hours.
A heat stable tracer containing protein which is prepared by:
- (a) labeling a mixture of tubercular proteins isolated from the growth media of Mycobacterium tuberculosis with a tracer and chromatographically separating a labeled tubercular protein having a molecular weight of between 20,000 and 30,000, stability over a pH range of from 3.0-9.0 stability over a temperature range of 4.degree. C.-250.degree. C. and well-defined mobility on starch gel electrophoresis.
A process for preparing a tracer-containing protein purified from Mycobacterium tuberculosis which comprises:
- labeling a mixture of tubercular proteins isolated from the growth media of Mycobacterium tuberculosis;
- chromatographing from said mixture of labeled tubercular proteins, a labeled, charcoal-adsorbable protein having a molecular weight of between 20,000 and 30,000, which is stable over a pH range of from 3.0 to 9.0 and over a temperature range of 4.degree. C.-250.degree. C. and has well-defined mobility characteristics on starch gel electrophoresis.
Dependencies are inferred from the claim text.