Diagnostic reagents

Abstract

There is provided a diagnostic reagent useful to determine whether an animal has a tuberculosis infection or has been exposed to a tuberculosis agent, for example a Mycobacterium . The reagent is useful to distinguish between such an animal and an animal which has been vaccinated against a tuberculosis infection.

1 . A diagnostic reagent comprising a polypeptide comprising ammo acid sequence SEQ ID NO:7 or a functional variant thereof, characterised in that the reagent elicits a negative diagnostic assay result when a tuberculosis infection assay is carried out on a sample from an animal which has been vaccinated against infection by a tuberculosis agent. 
     
     
         2 - 17 . (canceled) 
     
     
         18 . A method of detecting a tuberculosis infection in an animal, or exposure of an animal to a tuberculosis agent, comprising the steps of
 (i) contacting a population of cells from the animal with at least one diagnostic reagent consisting of amino acid sequence SEQ ID NO:7, or consisting of a polypeptide having 20-65 amino acids and comprising SEQ ID NO:7, or comprising a polypeptide which is a functional variant of SEQ ID NO:7 having 2, 4, or 5 amino acids added to or deleted from the N- or C-terminus and/or having at least 85% sequence identity to SEQ ID NO:7, characterised in that the reagent elicits a negative diagnostic assay result when a tuberculosis infection assay is carried out on a sample from an animal which has been vaccinated against infection by a tuberculosis agent using the live attenuated  M. bovis  strain Bacille Calmette-Guerin (BCG); and   (ii) determining whether the cells of said population recognise the diagnostic reagent.   
     
     
         19 . The method according to  claim 18 , wherein the population of cells includes T-cells. 
     
     
         20 . The method according to  claim 18 , comprising a cell-mediated immunity 10 (CMI) assay. 
     
     
         21 . The method according to  claim 20 , wherein the CMI assay detects interferon gamma (IFN-γ). 
     
     
         22 . A method of detecting a tuberculosis infection in an animal, or exposure of an animal to a tuberculosis agent, comprising conducting a skin test on the animal using at least one diagnostic reagent consisting of amino acid sequence SEQ ID NO:7, or consisting of a polypeptide having 20-65 amino acids and comprising SEQ ID NO:7, or comprising a polypeptide which is a functional variant of SEQ ID NO:7 having 2, 4, or 5 amino acids added to or deleted from the N- or C-terminus and/or having at least 85% sequence identity to SEQ ID NO:7, characterised in that the reagent elicits a negative diagnostic assay result when a tuberculosis infection assay is carried out on a sample from an animal which has been vaccinated against infection by a tuberculosis agent using the live attenuated  M. bovis  strain Bacille Calmette-Guerin (BCG). 
     
     
         23 - 27 . (canceled) 
     
     
         28 . A nucleic acid encoding a polypeptide comprising at least one polypeptide comprising the amino acid sequence of any of SEQ ID NOs:1-10 and/or 50-69 or a functional variant thereof. 
     
     
         29 - 30 . (canceled) 
     
     
         31 . The method of  claim 18 , wherein said diagnostic reagent further comprises at least one of the amino acid sequences SEQ ID NO:1, 2, 3, 4, 5, 6 or 9 or functional variants thereof, or comprising all of SEQ ID NOs:1-9. 
     
     
         32 . The method of  claim 18 , wherein said diagnostic reagent further comprises a polypeptide of the amino acid sequence of SEQ ID NO:10 or a functional variant thereof. 
     
     
         33 . The method of  claim 18 , wherein said diagnostic reagent further comprises further comprising at least one of the amino acid sequences SEQ ID NOs:11-69 or functional variants thereof. 
     
     
         34 . The method of  claim 22 , wherein said diagnostic reagent further comprises at least one of the amino acid sequences SEQ ID NO:1, 2, 3, 4, 5, 6 or 9 or functional variants thereof, or comprising all of SEQ ID NOs:1-9. 
     
     
         35 . The method of  claim 22 , wherein said diagnostic reagent further comprises a polypeptide of the amino acid sequence of SEQ ID NO:10 or a functional variant thereof. 
     
     
         36 . The method of  claim 22 , wherein said diagnostic reagent further comprises further comprising at least one of the amino acid sequences SEQ ID NOs:11-69 or functional variants thereof.