Skip to main content
NCBI home page
Journal of Clinical Pathology logoLink to Journal of Clinical Pathology
. 1996 Aug;49(8):642–647. doi: 10.1136/jcp.49.8.642

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

P J Horner 1, M Ali 1, D Parker 1, J N Weber 1, D Taylor-Robinson 1, M O McClure 1
PMCID: PMC500607  PMID: 8881914

Abstract

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.

Full text

PDF
642

Images in this article

644Image
on p.644
645Image
on p.645
646Image
on p.646

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bavoil P., Stephens R. S., Falkow S. A soluble 60 kiloDalton antigen of Chlamydia spp. is a homologue of Escherichia coli GroEL. Mol Microbiol. 1990 Mar;4(3):461–469. doi: 10.1111/j.1365-2958.1990.tb00612.x. [DOI] [PubMed] [Google Scholar]
  2. Craig E. A. Chaperones: helpers along the pathways to protein folding. Science. 1993 Jun 25;260(5116):1902–1903. doi: 10.1126/science.8100364. [DOI] [PubMed] [Google Scholar]
  3. Hutchinson G. R., Taylor-Robinson D., Dourmashkin R. R. Growth and effect of chlamydiae in human and bovine oviduct organ cultures. Br J Vener Dis. 1979 Jun;55(3):194–202. doi: 10.1136/sti.55.3.194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Jackett P. S., Bothamley G. H., Batra H. V., Mistry A., Young D. B., Ivanyi J. Specificity of antibodies to immunodominant mycobacterial antigens in pulmonary tuberculosis. J Clin Microbiol. 1988 Nov;26(11):2313–2318. doi: 10.1128/jcm.26.11.2313-2318.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Morrison R. P., Belland R. J., Lyng K., Caldwell H. D. Chlamydial disease pathogenesis. The 57-kD chlamydial hypersensitivity antigen is a stress response protein. J Exp Med. 1989 Oct 1;170(4):1271–1283. doi: 10.1084/jem.170.4.1271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Morrison R. P., Su H., Lyng K., Yuan Y. The Chlamydia trachomatis hyp operon is homologous to the groE stress response operon of Escherichia coli. Infect Immun. 1990 Aug;58(8):2701–2705. doi: 10.1128/iai.58.8.2701-2705.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Taylor H. R., Johnson S. L., Schachter J., Caldwell H. D., Prendergast R. A. Pathogenesis of trachoma: the stimulus for inflammation. J Immunol. 1987 May 1;138(9):3023–3027. [PubMed] [Google Scholar]
  8. Thomas B. J., Reeve P., Oriel J. D. Simplified serological test for antibodies to Chlamydia trachomatis. J Clin Microbiol. 1976 Jul;4(1):6–10. doi: 10.1128/jcm.4.1.6-10.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Toye B., Laferrière C., Claman P., Jessamine P., Peeling R. Association between antibody to the chlamydial heat-shock protein and tubal infertility. J Infect Dis. 1993 Nov;168(5):1236–1240. doi: 10.1093/infdis/168.5.1236. [DOI] [PubMed] [Google Scholar]
  11. Wagar E. A., Schachter J., Bavoil P., Stephens R. S. Differential human serologic response to two 60,000 molecular weight Chlamydia trachomatis antigens. J Infect Dis. 1990 Oct;162(4):922–927. doi: 10.1093/infdis/162.4.922. [DOI] [PubMed] [Google Scholar]
  12. Watkins N. G., Hadlow W. J., Moos A. B., Caldwell H. D. Ocular delayed hypersensitivity: a pathogenetic mechanism of chlamydial-conjunctivitis in guinea pigs. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7480–7484. doi: 10.1073/pnas.83.19.7480. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Pathology are provided here courtesy of BMJ Publishing Group

RESOURCES