Antigen capture ELISA for the heat shock protein (hsp60) of Chlamydia trachomatis

被引:3
作者
Horner, PJ
Ali, M
Parker, D
Weber, JN
TaylorRobinson, D
McClure, MO
机构
[1] ST MARYS COLL,SCH MED,DEPT GENITOURINARY MED & COMMUNICABLE DIS,JEFFERISS RES TRUST LABS,LONDON W2 1NY,ENGLAND
[2] MRC,SEXUALLY TRANSMITTED DIS RES GRP,LONDON W1N 4AL,ENGLAND
[3] MUREX DIAGNOST LTD,DEPTFORD DA1 5LR,KENT,ENGLAND
关键词
Chlamydia trachomatis; heat shock protein; ELISA; antibody;
D O I
10.1136/jcp.49.8.642
中图分类号
R36 [病理学];
学科分类号
100104 ;
摘要
Aims-To develop an indirect ELISA using the heat shock protein (hsp60) of Chlamydia trachomatis as antigen. Methods-The hsp60 gene was amplified by PCR, expressed in the vector pDEV-107 and transformed into Escherichia coli. The recombinant protein, expressed as a beta-galactosidase fusion product, was captured onto a solid phase using a monoclonal antibody directed against beta-galactosidase, Following incubation with goat anti-human antibody conjugated to peroxidase and colour development on addition of peroxidase substrate, antibody recognition of antigen was quantified by optical density at 492 nm. Results-A sensitive and relatively specific ELISA to detect hsp60 has been produced, which can be exploited to determine the antibody response to C trachomatis hsp60. Conclusions-This assay will permit the future investigation of the immunopathogenesis of persistent inflammation following C trachomatis infection.
引用
收藏
页码:642 / 647
页数:6
相关论文
共 15 条
  • [1] A SOLUBLE 60-KILODALTON ANTIGEN OF CHLAMYDIA-SPP IS A HOMOLOG OF ESCHERICHIA-COLI GROEL
    BAVOIL, P
    STEPHENS, RS
    FALKOW, S
    [J]. MOLECULAR MICROBIOLOGY, 1990, 4 (03) : 461 - 469
  • [2] CLONING AND SEQUENCE OF THE GENE FOR HEAT-SHOCK PROTEIN-60 FROM CHLAMYDIA-TRACHOMATIS AND IMMUNOLOGICAL REACTIVITY OF THE PROTEIN
    CERRONE, MC
    MA, JJ
    STEPHENS, RS
    [J]. INFECTION AND IMMUNITY, 1991, 59 (01) : 79 - 90
  • [3] CHAPERONES - HELPERS ALONG THE PATHWAYS TO PROTEIN-FOLDING
    CRAIG, EA
    [J]. SCIENCE, 1993, 260 (5116) : 1902 - 1903
  • [4] Dunn, 1986, ELECTROPHORESIS 86, P315
  • [5] HUTCHINSON GR, 1979, BRIT J VENER DIS, V55, P194
  • [6] SPECIFICITY OF ANTIBODIES TO IMMUNODOMINANT MYCOBACTERIAL ANTIGENS IN PULMONARY TUBERCULOSIS
    JACKETT, PS
    BOTHAMLEY, GH
    BATRA, HV
    MISTRY, A
    YOUNG, DB
    IVANYI, J
    [J]. JOURNAL OF CLINICAL MICROBIOLOGY, 1988, 26 (11) : 2313 - 2318
  • [7] CHLAMYDIAL DISEASE PATHOGENESIS - THE 57-KD CHLAMYDIAL HYPERSENSITIVITY ANTIGEN IS A STRESS RESPONSE PROTEIN
    MORRISON, RP
    BELLAND, RJ
    LYNG, K
    CALDWELL, HD
    [J]. JOURNAL OF EXPERIMENTAL MEDICINE, 1989, 170 (04) : 1271 - 1283
  • [8] THE CHLAMYDIA-TRACHOMATIS HYP OPERON IS HOMOLOGOUS TO THE GROE STRESS RESPONSE OPERON OF ESCHERICHIA-COLI
    MORRISON, RP
    SU, H
    LYNG, K
    YUAN, Y
    [J]. INFECTION AND IMMUNITY, 1990, 58 (08) : 2701 - 2705
  • [9] Sambrook J., 2002, MOL CLONING LAB MANU
  • [10] TAYLOR HR, 1987, J IMMUNOL, V138, P3023