Deleted sequences in m. bovis bcg/m. bovis or m. tuberculosis, method for detecting mycobacteria using said sequences and vaccines

Abstract

The invention concerns the isolation of nucleotide and peptide sequences in particular for differentiating, in diagnostic terms, an immunisation resulting from BCG vaccination of an infection by M. tuberculosis . Said sequences are either M. bovis BCG/ M. bovis specific or M. tuberculosis specific. The invention also concerns a method for detecting said sequences, a method for detecting antibodies generated by the products expressing said sequences as well as kits for implementing said methods. Finally, the invention concerns novel vaccines.

1 - 25 . (canceled) 
     
     
         26 . An isolated nucleic acid which is a primer or molecular hybridization probe comprising sequence from  M. tuberculosis , wherein the  M. tuberculosis  sequence in the primer or probe consists of a sequence of ORF Rv2075c which is absent from  M. microti  strain OV254 due to deletion of RD9. 
     
     
         27 . The isolated nucleic acid of  claim 26 , further comprising a radioactive label. 
     
     
         28 . The isolated nucleic acid of  claim 26 , further comprising a non-radioactive label. 
     
     
         29 . The isolated nucleic acid of  claim 28 , wherein the non-radioactive label comprises an enzyme. 
     
     
         30 . The isolated nucleic acid of  claim 28 , wherein the non-radioactive label comprises a fluorescent label. 
     
     
         31 . The isolated nucleic acid of  claim 26 , wherein the sequence of the isolated nucleic acid consists essentially of the  M. tuberculosis  sequence. 
     
     
         32 . The isolated nucleic acid of  claim 26 , wherein the sequence of the isolated nucleic acid consists of the  M. tuberculosis  sequence. 
     
     
         33 . The isolated nucleic acid of  claim 26 , which was produced by a process comprising amplifying the isolated nucleic acid from  M. tuberculosis  genomic DNA,  M. tuberculosis  RNA, or  M. tuberculosis  cDNA. 
     
     
         34 . The isolated nucleic acid of  claim 26 , wherein the isolated nucleic acid is a molecular hybridization probe. 
     
     
         35 . The molecular hybridization probe of  claim 34 , which is contained in a recombinant plasmid. 
     
     
         36 . The isolated nucleic acid of  claim 26 , wherein the isolated nucleic acid is a primer. 
     
     
         37 . A pair of primers according to  claim 36 . 
     
     
         38 . An amplification reaction mixture comprising a pair of primers according to  claim 37 . 
     
     
         39 . The amplification reaction mixture of  claim 38 , further comprising genomic DNA or cDNA of  M. tuberculosis.    
     
     
         40 . The amplification reaction mixture of  claim 38 , further comprising genomic DNA or cDNA of a mycobacterium other than  M. tuberculosis.    
     
     
         41 . A method of characterizing a nucleic acid of a mycobacterium in a biological sample, comprising:
 (i) isolating DNA from the biological sample or preparing cDNA from RNA of the biological sample; and   (ii) performing an assay on the DNA or cDNA for detecting Rv2075c sequence which is present in  M. tuberculosis  but absent from  M. microti  strain OV254 due to deletion of RD9.   
     
     
         42 . The method of  claim 41 , wherein the assay comprises a hybridization assay which optionally comprises an electrophoresis step. 
     
     
         43 . The method of  claim 41 , wherein the assay comprises an amplification assay. 
     
     
         44 . The method of  claim 41 , wherein the assay comprises a PCR assay. 
     
     
         45 . The method of  claim 41 , wherein the biological sample comprises serum, blood, a biopsy, bronchoalveolar fluid, or pleural fluid of a human or animal.