Binding assay for the detection of mycobacteria
AssigneeMontefiore Medical Center
InventorEugene W. Straus
Life of this patent
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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 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. 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.
Claims26
A kit for determining the presence of Mycobacteria comprising a carrier being compartmented to receive at least two vials and to maintain said vials in close confinement, which comprises:
- a first vial containing a tracer containing molecule wherein said tracer containing molecule is a heat stable charcoal-adsorbable, radiolabeled protein obtained from the growth media of Mycobacterium tuberculosis, said protein having a molecular weight of between 20,000 and 30,000, immunochemical stability from 4.degree. C.-250.degree. C. and from pH 3.0 to 9.0, and exhibiting comigration towards the anode in the presence of bovine serum albumin and borate buffer at pH of 8.6 with an applied voltage of 80 volts on starch gel electrophoresis for 12 hours and a second vial containing antibodies raised against or reacting with tubercular proteins isolated from a bacterium of the genus Mycobacterium, and which have reversible binding affinity towards said protein in said first vial wherein the tracer containing molecule and antibodies are present in amounts sufficient to perform an immunoassay for Mycobacteria.
A kit comprising a carrier being compartmented to receive at least two vials and to maintain said vials in a close confinement, which comprises:
- a first vial containing a complex of (A) a tracer containing molecule which is a radiolabled, charcoal-adsorbable, heat-stable protein obtained from the growth media of Mycobacterium tuberculosis with a molecular weight of between 20,000-30,000, which is immunochemically stable from 4.degree. C. to 250.degree. C. and over a pH range of 3.0 to 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 with;
- (B) antibodies having reversible affinity towards said protein and which antibodies have been raised against tubercular proteins isolated from a bacterium of the genus Mycobacterium wherein the tracer containing molecule and the antibodies are present in amounts sufficient to perform an immunoassay for Mycobacteria and a second vial containing a buffer.
A method for detecting the presence of Mycobacteria in a fluid or tissue, which comprises:
- mixing said fluid or tissue containing a secretory product of Mycobacteria with (a) insoluble carrier particles having attached thereon a protein obtained from the growth media of Mycobacterium tuberculosis of molecular weight of between 20,000-30,000 which is heat-stable, charcoal-adsorbable and which is immunochemically stable from 4.degree. C. to 250.degree. C. and from pH 3.0-9.0 and exhibiting comigration towards the anode in the presence of bovine serum albumin and borate buffer of a pH of 8.6 with an applied voltage of 80 volts on starch gel electrophoresis for 12 hours;
- and with (b) antibodies raised against tuberculosis protein isolated from a bacterium of the genus Mycobacterium;
- and
- detecting the inhibition of agglutination of said insoluble carrier particles.
A heat stable, charcoal-adsorbable protein purified from the growth media of Mycobacterium tuberculosis, having a molecular weight between 20,000 and 30,000, immunochemical stability over a temperature range of 4.degree. C.-250.degree. C., a pH range of from 3.0 to 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 kit comprising a carrier being compartmented to receive at least two vials and to maintain said vials in close confinement, which comprises:
- a first vial containing a heat stable, charcoal adsorbable protein purified from the growth media of Mycobacterium tuberculosis having a molecular weight between 20,000 and 30,000, which is immunochemically stable over a temperature range of 4.degree. C.-250.degree. C. and over a pH range of 3.0-9.0 and which, when labeled, exhibits 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;
- and
- a second vial containing antibodies raised against or reacting with tubercular proteins isolated from a bacterium of the genus Mycobacterium, and which has reversible binding affinity towards said protein in said first vial wherein the protein and antibodies are present in amounts sufficient to perform an immunoassay for Mycobacteria.
Dependencies are inferred from the claim text.