Patent
There is provided a diagnostic reagent for use in the detection of M. bovis or M. tuberculosis infection in an animal, comprising a peptide which has an epitope from Mycobacterium bovis hypothetic protein Mb3645c (SEQ ID NO: 1) or an epitope from a polypeptide having at least 76% identity with SEQ ID NO: 1.
1. An isolated peptide consisting of the amino acid sequence of SEQ ID NO: 9, 10, 11, 12, or 13, or a functional variant peptide thereof having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 9, 10, 11, 12, or 13.
2. A composition comprising at least two, three, four, or five of the isolated peptides or variants thereof of claim 1 .
3. A cell-mediated immunity (CMI) assay comprising at least one of the peptides of claim 1 .
4. A diagnostic kit comprising at least one of the peptides or variants thereof of claim 1 .
5. The diagnostic kit of claim 4 , wherein said kit further comprises at least one peptide consisting of the amino acid sequence of SEQ ID NO: 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25.
6. The diagnostic kit of claim 4 , wherein said kit comprises at least two, three, four, or five of said isolated peptides or variants thereof.
7. A method for diagnosing in a host an infection of or exposure to Mycobacterium bovis or Mycobacterium tuberculosis comprising
(a) contacting a population of cells from the host with an isolated peptide consisting of the amino acid sequence of SEQ ID NO: 1, 9, 10, 11, 12, or 13, or a functional variant peptide thereof having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 1, 9, 10, 11, 12, or 13; and
(b) determining whether the cells of said cell population recognize the peptide.
8. The method of claim 7 , wherein recognition of the peptide is able to detect a Mycobacterium bovis or Mycobacterium tuberculosis infection in a mammal.
9. The method of claim 7 , wherein said detecting is not susceptible to diagnosis by using an ESAT-6/CFP-10 peptide cocktail.
10. The method of claim 7 , wherein the population of cells includes T-cells.
11. The method of claim 7 , wherein recognition of the peptide is able to differentiate between an Mycobacterium bovis or Mycobacterium tuberculosis infected mammal and a mammal vaccinated against infection by Mycobacterium bovis or Mycobacterium tuberculosis.
12. The method of claim 11 , wherein the mammal vaccinated against Mycobacterium bovis and/or Mycobacterium tuberculosis has been vaccinated with the live attenuated vaccine Mycobacterium bovis bacillus Calmette-Guerin (BCG).
13. The method of claim 7 , further comprising a cell-mediated immunity (CMI) assay.
14. The method of claim 13 , wherein the CMI assay detects interferon gamma.