Use of amino acid sequences from mycobacterium tuberculosis or corresponding nucleic acids for diagnosis and prevention of tubercular infection, diagnostic kit and vaccine therefrom

A method for detecting a Mycobacterium species infection in a test subject, comprising:

• incubating in vitro a biological sample that comprises lymphocytes from the test subject with at least one protein or peptide that is selected from:
• (i) an isolated protein that comprises an amino acid sequence selected from the amino acid sequence set forth as Rv0023 (SEQ ID NO:1), Rv0182c (SEQ ID NO:2), Rv0290 (SEQ ID NOS:4 and 5), Rv0601c (SEQ ID NOS:6-10), Rv0647c (SEQ ID NOS:11-12), Rv0724A (SEQ ID NO:13), Rv0890c (SEQ ID NOS:14-20), Rv1251c (SEQ ID NOS:21-22), Rv1398c (SEQ ID NO:23), Rv1478 (SEQ ID NO:24), Rv1497 (SEQ ID NOS:25-26), Rv1575 (SEQ ID NO:27), Rv1578c (SEQ ID NO:28), Rv1899c (SEQ ID NOS:29-31), Rv2137c (SEQ ID NO:32), Rv2333c (SEQ ID NOS:33-35), Rv2548 (SEQ ID NOS:36-37), Rv2557 (SEQ ID NOS:38-40), Rv2816c (SEQ ID NOS:41-42), Rv2990 (SEQ ID NOS:43-47), Rv3094c (SEQ ID NOS:48-51), Rv3107c (SEQ ID NO:52), Rv3188 (SEQ ID NO:53), Rv3239c (SEQ ID NOS:54-61), Rv3296 (SEQ ID NOS:62-63), Rv3425 (SEQ ID NOS:64-66), Rv3446c (SEQ ID NOS:67-69), Rv3479 (SEQ ID NOS:70, 71 and 87), Rv3482c (SEQ ID NOS:72-73), and Rv3780 (SEQ ID NO:74), derived from a Mycobacterium species and comprising at least one T-cell epitope,
• (ii) a homolog of the protein as defined in (i) having an amino acid sequence with at least 80% similarity to said protein after optimal alignment,
• (iii) a peptide fragment of the protein as defined in (i) or (ii) having a T-cell epitope, or a chemical analog thereof, and
• (iv) a Mycobacterium protein, or a fragment thereof or a chemical analog thereof, said Mycobacterium protein being selected from:
• (a) ESAT,
• (b) CFP10,
• (c) TB7.7, and
• (d) PPD,
• wherein said step of incubating is for a time and under conditions sufficient to stimulate the lymphocytes to produce an effector molecule;
• and
• detecting a test level of the effector molecule, wherein detection of a test level that is higher than a control level that is detected following incubation of control lymphocytes from a healthy control subject, indicates that the test subject is infected with, or has been previously exposed to, the Mycobacterium species.

An isolated protein selected from:

• (a) an isolated protein that comprises an amino acid sequence selected from the amino acid sequence set forth as Rv0023 (SEQ ID NO:1), Rv0182c (SEQ ID NO:2), Rv0290 (SEQ ID NOS:4 and 5), Rv0601c (SEQ ID NOS:6-10), Rv0647c (SEQ ID NOS:11-12), Rv0724A (SEQ ID NO:13), Rv0890c (SEQ ID NOS:14-20), Rv1251c (SEQ ID NOS:21-22), Rv1398c (SEQ ID NO:23), Rv1478 (SEQ ID NO:24), Rv1497 (SEQ ID NOS:25-26), Rv1575 (SEQ ID NO:27), Rv1578c (SEQ ID NO:28), Rv1899c (SEQ ID NOS:29-31), Rv2137c (SEQ ID NO:32), Rv2333c (SEQ ID NOS:33-35), Rv2548 (SEQ ID NOS:36-37), Rv2557 (SEQ ID NOS:38-40), Rv2816c (SEQ ID NOS:41-42), Rv2990 (SEQ ID NOS:43-47), Rv3094c (SEQ ID NOS:48-51), Rv3107c (SEQ ID NO:52), Rv3188 (SEQ ID NO:53), Rv3239c (SEQ ID NOS:54-61), Rv3296 (SEQ ID NOS:62-63), Rv3425 (SEQ ID NOS:64-66), Rv3446c (SEQ ID NOS:67-69), Rv3479 (SEQ ID NOS:70, 71 and 87), Rv3482c (SEQ ID NOS:72-73), and Rv3780 (SEQ ID NO:74), or that consists of the amino acid sequence LAWITAVVPGLMV (SEQ ID NO:85) or GEIIFISGRLNG (SEQ ID NO:86), said isolated protein comprising a mycobacterial epitope that is recognized by T-cells, and
• (b) the isolated protein of (a) which consists of an amino acid sequence selected from TAWITAVVPGLMV (SEQ ID NO:24), AVIVRSELLTQYL (SEQ ID NO:22), GSVRQLPSVLKPPLITLRTLTLSG (SEQ ID NO:71), RPVRRVLLFVVPSSGPAP (SEQ ID NO:70), GEIIFISGRLNGaa (SEQ ID NO:
• 13), ELMARAAVLGSAH (SEQ ID NO:21), LAWITAVVPGLMV (SEQ ID NO:85), GEIIFISGRLNG (SEQ ID NO:86), and SALLRRLSTCPPES (SEQ ID NO:87).