Patent HU-9903876-A2 (HU)
Title: Compounds and methods for diagnosis of tuberculosis
Inventor: CAMPOS-NETO ANTONIO
IPC: C12Q1/68
Filed: 1997-10-07
Abstract: A new polypeptide (A) comprises an antigenic portion of a soluble Mycobacterium tuberculosis (MT) antigen, or a variant of the antigen that differs only in conservative substitutions and/or modifications, the antigen has an N-terminal sequence selected from: (a) Asp-Pro-Val-Asp-Ala-Val-Ile-Asn-Thr-Thr-Cys-Asn-Tyr-Gly-Gln-Val-Val-Ala-Ala-Leu (I); (b) Ala-Val-Glu-Ser-Gly-Met-Leu-Ala-Leu-Gly-Thr-Pro-Ala-Pro-Ser (II); (c) Ala-Ala-Met-Lys-Pro-Arg-Thr-Gly-Asp-Gly-Pro-Leu-Glu-Ala-Ala-Lys-Glu-Gly-Arg (III); (d) Tyr-Tyr-Trp-Cys-Pro-Gly-Gln-Pro-Phe-Asp-Pro-Ala-Trp-Gly-Pro (IV); (e) Asp-Ile-Gly-Ser-Glu-Ser-Thr-Glu-Asp-Gln-Gln-Xaa-Ala-Val (V); (f) Ala-Glu-Glu-Ser-Ile-Ser-Thr-Xaa-Glu-Xaa-Ile-Val-Pro (VI); (g) Asp-Pro-Glu-Pro-Ala-Pro-Pro-Val-Pro-Thr-Thr-Ala-Ala-Ser-Pro-Pro-Ser (VII); (h) Ala-Pro-Lys-Thr-Tyr-Xaa-Glu-Glu-Leu-Lys-Gly-Thr-Asp-Thr-Gly; (VIII); (i) Asp-Pro-Apa-Ser-Ala-Pro-Asp-Val-Pro-Thr-Ala-Ala-Gln-Leu-Thr-Ser-Leu-Leu-Asn-Ser-Leu-Ala-Asp-Pro-Asn-Val-Ser-Phe-Ala-Asn (I X); (j) Ala-Pr o-Glu-Ser-Gly-Ala-Gly-Leu-Gly-Leu-Gly-Gly-Thr-Val-Gln-Ala-Gly (X); where Xaa may be any amino acid. Also claimed are: (1) a polypeptide comprising an immunogenic portion of an MT antigen or a variant of the antigen that differs only in conservative substitutions and/or modifications, where the antigen has an N-terminal sequence selected from: (a) Asp-Pro-Pro-Asp-Pro-His-Gln-Xaa-Asp-Met-Thr-Lys-Gly-Tyr-Tyr-Pro-Gly-Gly-Arg-Arg-Xaa-Phe (XI); and (b) Xaa-Tyr-Ile-Ala-Tyr-Xaa-Thr-Thr-Ala Gly-Ile-Val-Pro-Gly-Lys-Ile-Asn-Val-His-Leu-Val (XII); (2) a polypeptide comprising an antigenic portion of a soluble MT antigen, or a variant of the antigen that differs only in conservative substitutions and/or modifications, where the antigen comprises an amino acid sequence encoded by DNA of one of the 766, 752, 447, 604, 633, 1362, 1458, 862, 622, 1771, 1058, 913, 1872, 1482, 876, 1021, 321, 373, 352, 726, 580, 999, 507 and 500 bp sequences given in the specification, or the complements of the sequences, or DNA sequences that hybridise to these sequences or a complement under moderately stringent conditions; (3) a polypeptide as in (2) except that it is encoded by one of the 272, 317, 182, 308, 267, 1539, 851, 254, 1227, 181, 290, 34, 155, 53, 41, 132, 702, 298, 1058, 327, 170, 127, 81, 149, 355, 999, 882, 815, 264, 1171, 227, 304, 1439, 329, 80, 392, 535, 690, 407, 468, 219, 494, 220, 388, 400, 538, 239, 985, 2138 and 2367 bp sequences given in the specification, the complements of the sequences, and DNA sequences that hybridise to a these sequences or a complement under moderately stringent conditions; (4) a DNA molecule (I) comprising a nucleotide sequence encoding a polypeptide as in (A) or as described in (1), (2) or (3); (5) an expression vector comprising (I); (5) a host cell transformed with an expression vector as in (5); (6) a method for detecting M. tuberculosis comprising contacting a biological sample with a peptide (A) or as described in (1), (2) or (3) and detecting antibodies that bind to these peptides; (7) a method as in (6) except that the peptide is one of a 15 or 15 aa sequence given in the specification; (8) a method as in (6) except that the peptide is encoded by one of the 813, 1200, 1155, 1152, 136, 1777, 324, 1338, 321, 492, 536, 2072,1923, 1055, 359, 350, 811, 966 and 2852 sequences given in the specification. (9) a method for detecting M. tuberculosis comprises using PCR and two oligonucleotide primers specific to the (I) or the sequences described in (6) to generate a detectible amplicon. (10) a method for detecting M. tuberculosis comprises using probes that hybridise specifically to the DNA sequences as described in (6);(11) a method for detecting M. tuberculosis comprises: (a) contacting a sample with a binding agent that can bind (A) or a peptide as described in (1), (2), (3) or (6), or as described in the two 15 aa sequences given in the specification; and (b) detecting the bound peptide; (12) a polyclonal antibody that binds to (A) or a peptide as in (1), (2) or (3); (13) a fusion protein comprising (A)