Biomarkers for early diagnosis and differentiation of mycobacterial infection

Abstract

Mycobacterial-specific biomarkers and methods of using such biomarkers for diagnosis of mycobacterial infection in a mammal are disclosed.

We claim: 
     
         1 . A method for diagnosis of mycobacterial infection in a mammal, the method comprising the steps of
 a) obtaining a sample from the mammal;   b) testing the sample for the concentration level of at least one mycobacterial-specific biomarker and comparing the level of the biomarker against the level detected in an uninfected mammalian sample; and   c) determining the infection status of the mammal.   
     
     
         2 . The method of  claim 1 , wherein the method is used for early diagnosis and detection of mycobacterial infection in a mammal. 
     
     
         3 . The method of  claim 1 , wherein the testing is via ELISA assay for antibodies formed against the biomarker. 
     
     
         4 . The method of  claim 1 , wherein the sample is a blood sample. 
     
     
         5 . The method of  claim 1 , wherein the mammal is selected from the group consisting of bubaline, elephantine, musteline, pardine, phocine, rhinocerine, caprine, hircine, leonine, leporine, lupine, lyncine, murine, rusine, tigrine, ursine, vulpine, zebrine, vespertilionine, porcine, bovine, equine, swine, elaphine, ovine, caprine, camelidae, feline, cervine, primate, human and canine mammals. 
     
     
         6 . The method of  claim 1  wherein the mammal is selected from the group of pig, cow, human, rodent, sheep, goat and deer. 
     
     
         7 . The method of  claim 1 , wherein the mammal is selected from the group consisting of cow, sheep and goat. 
     
     
         8 . The method of  claim 1 , wherein the mammal is a cow. 
     
     
         9 . The method of  claim 1 , wherein the mycobacterial-specific biomarker comprises at least one member selected from the group consisting of gene sequences Q73SF4, Q73Y73, Q73ZE6, Q73SL7, Q73VK6, Q73XZ0, Q740D1 and Q73UE0 and expression products derived thereof. 
     
     
         10 . The method of  claim 1 , wherein the mycobacterial-specific biomarker comprises at least one member selected from the group consisting of gene sequences Q73VL6, Q73YW9, Q741L4, Q744E5, Q73YP5, Q73WE5, Q73U21, Q73UH9, Q741M5, Q742F4, and Q73SU6 and expression products derived thereof. 
     
     
         11 . The method of  claim 1 , wherein the biomarker comprises a protein having at least 50%, 60%, 70%, 80%, or 90% of the amino acids of  M. paratuberculosis  which are not conserved in  M. bovis  as exemplified by the amino acid sequence difference between  M. paratuberculosis  and  M. bovis  in  FIG. 1 . 
     
     
         12 . A method for differentiating a vaccinated mammal from a non-vaccinated mammal or from an infected mammals, the method comprising the steps of
 a) obtaining a sample from the test mammal;   b) testing the sample for the concentration level of at least one of mycobacterial specific markers and comparing the level of the markers with that detected in an uninfected animal; and   c) determining the status of the mammal.   
     
     
         13 . The method of  claim 12 , wherein the mammals are vaccinated with a  mycobacterium  mutant vaccine. 
     
     
         14 . The method of  claim 13 , wherein the  mycobacterium  mutant vaccine comprises at least one mutation in at least one gene sequence encoding global gene regulators (GGRs) selected from the group consisting of sigH, sigL and LipN. 
     
     
         15 . The method of  claim 12 , wherein the marker comprises at least one member selected from the group consisting of Q73SF4, Q73Y73, Q73ZE6, Q73SL7, Q73VK6, Q73XZ0, Q740D1 and Q73UE0 and expression products derived thereof. 
     
     
         16 . The method of  claim 12 , wherein the marker comprises at least one member selected from the group consisting of Q73VL6, Q73YW9, Q741L4, Q744E5, Q73YP5, Q73WE5, Q73U21, Q73UH9, Q741M5, Q742F4, and Q73SU6 and expression products derived thereof. 
     
     
         17 . The method of  claim 12 , wherein the marker comprises a protein having at least 50%, 60%, 70%, 80%, or 90% of the amino acid sequence of  M. paratuberculosis  which is not conserved in  M. bovis , as exemplified by the amino acid difference between  M. paratuberculosis  and  M. bovis  in  FIG. 1 . 
     
     
         18 . A method for differentiating a  M. ap  infected mammal from a  M. bovis  infected mammal, the method comprising the steps of
 a) obtaining a sample from the test mammal;   b) testing the sample for the concentration level of at least one of mycobacterial-specific markers and comparing the level of the markers with that detected in a  M. bovis  infected mammal; and   c) determining the status of the mammal.   
     
     
         19 . The method of  claim 18 , wherein the mycobacterial-specific markers comprise a protein having at least 50%, 60%, 70%, 80%, or 90% of the amino acid sequence of  M. paratuberculosis  which is not conserved in  M. bovis.    
     
     
         20 . A set of biomarkers for early diagnosis and differentiation of mycobacterial infection, the biomarkers comprising:
 at least one member selected from the group consisting of Q73SF4, Q73Y73, Q73ZE6, Q73SL7, Q73VK6, Q73XZ0, Q740D1 and Q73UE0 and expression products derived thereof.   
     
     
         21 . The set of biomarkers in  claim 20 , wherein the biomarkers comprising at least two members selected from the group consisting of Q73SF4, Q73Y73, Q73ZE6, Q73SL7, Q73VK6, Q73XZ0, Q740D1 and Q73UE0 and expression products derived thereof. 
     
     
         22 . A set of biomarkers for early diagnosis and differentiation of mycobacterial infection, the biomarkers comprising:
 at least one member selected from the group consisting of Q73VL6, Q73YW9, Q741L4, Q744E5, Q73YP5, Q73WE5, Q73U21, Q73UH9, Q741M5, Q742F4, and Q73SU6 and encoded genes or expression products derived thereof.   
     
     
         23 . The set of biomarkers in  claim 22 , wherein the biomarkers comprising at least two members selected from the group consisting of Q73VL6, Q73YW9, Q741L4, Q744E5, Q73YP5, Q73WE5, Q73U21, Q73UH9, Q741M5, Q742F4, and Q73SU6 and encoded genes or expression products derived thereof. 
     
     
         24 . A set of biomarkers for early diagnosis and differentiation of mycobacterial infection, the biomarkers comprising a protein having at least 50%, 60%, 70%, 80%, 90%, 95% or 100% of the amino acid sequence of  M. paratuberculosis  which is not conserved in  M. bovis.