# US 5,707,819 A · Diagnosis of Mycobacterium bovis infection > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 5,707,819 A - Document ID: US5707819A - Status: Granted - Kind: A - Application number: 08/454,746 - Filed: 1995-05-31 - Granted: 1998-01-13 - Earliest claimed: 1988-03-31 - Assignees: Commonwealth Scientific and Industrial Research Organisation - Inventors: Paul Richard Wood; Anthony John Radford; Theodora Fifis - Web page: https://patentgrep.com/patent/US5707819A ## CPC - C07K 14/35: from Mycobacteriaceae (F) - C07K 16/1289: Mycobacteriaceae (F) - Y10S 436/811: Test for named disease, body condition or organ function - Y10S 530/82: Proteins from microorganisms - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - Y10S: TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS ## Abstract A method for the diagnosis of Mycobacterium bovis infection in a susceptible animal, comprises detection in said animal of antibodies against the MPB-70 protein of M.bovis and/or the detection of a cell-mediated immune response of said animal to the said MPB-70 protein. Also disclosed is a recombinant DNA molecule corresponding to all or portion of the M.bovis DNA sequence coding for the MPB-70 protein or a polypeptide having the antigenicity of MPB-70 protein, or degenerate forms thereof, as well as recombinant MPB-70 protein or polypeptide and a process for the preparation thereof. ## Claims ### Claim 1 A method for the detection of Mycobacterium bovis infection in a susceptible animal, which comprises detecting in said animal an immune response to MPB-70 protein of M.bovis having the amino acid sequence of FIG. 3 by means of an assay selected from the group consisting of an assay for MPB-70 recognizing antibodies and an assay for a cell-mediated immune response; said assay using said MPB-70 protein or an antigenic polypeptide derived therefrom reactive with affinity-purified antibody to M. bovis as antigen, and said MPB-70 protein or antigenic polypeptide derived therefrom being substantially free of other M.bovis proteins. ### Claim 2 (depends on 1) A method according to claim 1, wherein said animal is selected from the group consisting of cattle, deer, badgers, possums, pigs and camels. ### Claim 3 (depends on 1) A method according to claim 1, wherein a cell-mediated immune response of said animal is detected by an in vivo caudal fold skin test using said MPB-70 protein or an antigenic polypeptide derived therefrom as an antigen. ### Claim 4 (depends on 1) A method according to claim 1, wherein a cell-mediated immune response of said animal is detected by an in vivo assay using said MPB-70 protein or antigenic polypeptide derived therefrom as antigen. ### Claim 5 (depends on 4) A method according to claim 4, wherein said in vitro assay is a lymphocyte proliferation assay or an assay based on release of gamma interferon or interleukin.sup.2. ### Claim 6 A kit for the detection of Mycobacterium bovis infection in a susceptible animal, which comprises means for the detection in said animals of an immune response to MPB-70 protein of M.bovis having the amino acid sequence of FIG. 3 by means of an assay selected from the group consisting of an assay for MPB-70 recognizing antibodies and an assay for cell mediated immune response, said kit comprising said MPB-70 protein or an antigenic polypeptide derived therefrom reactive with affinity-purified antibody to M.bovis as antigen, and said MPB-70 protein or antigenic polypeptide derived therefrom being substantially free of other M.bovis proteins. ### Claim 7 A method for the detection of Mycobacterium bovis infection in a susceptible animal which comprises detecting in said animal an immune response to MPB-70 protein of M.bovis having the amino acid sequence of FIG. 3; said immune response being detected by contacting a serum sample from said animal with an antigen selected from the group consisting of said MPB-70 protein and antigenic polypeptides derived therefrom reactive with affinity-purified antibody to M.bovis; and detecting binding of antibodies in said sample with said antigen to indicate the presence of said antibodies in said sample; said MPB-70 protein or antigenic polypeptide derived therefrom being substantially free of other M.bovis proteins. ### Claim 8 A method for the detection of Mycobacterium bovis infection in a susceptible animal which comprises detecting in said animal an immune response to MPB-70 protein of M.bovis having the amino acid sequence of FIG. 3; said immune response being detected by contacting a serum sample from said animal with an antigen selected from the group consisting of said MPB-70 protein and antigenic polypeptides derived therefrom reactive with affinity-purified antibody to M.bovis; and detecting binding of antibodies in said sample with said antigen by competition with binding of labelled antibody to said MPB-70 protein to indicate the presence of said antibodies in said sample; said MPB-70 protein or antigenic polypeptide derived therefrom being substantially free of other M.bovis proteins. ### Claim 9 MPB-70 protein of M.bovis having the amino acid sequence of FIG. 3, or an antigenic polypeptide derived therefrom reactive with affinity-purified antibody to M.bovis, said MPB-70 protein or antigenic polypeptide derived therefrom being substantially free of other M.bovis proteins. ### Claim 10 (depends on 9) A method for the preparation of MPB-70 protein of claim 9, which comprises purification of an M.bovis culture filtrate by chromatofocussing. ### Claim 11 (depends on 10) A method according to claim 10, wherein said chromatofocusing step is followed by further purification by gel filtration.