Antigen capture ELISA for the heat shock protein (hsp60) of Chlamydia trachomatis.
OBJECTIVE
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.
Horner
P J
PJ
Department of Genito-urinary Medicine and Communicable Diseases, Imperial College School of Medicine at St Mary's, London.
Ali
M
M
Parker
D
D
Weber
J N
JN
Taylor-Robinson
D
D
McClure
M O
MO
D015153:Blotting, Western; D018834:Chaperonin 60 / Q000097:blood*; D002690:Chlamydia Infections / Q000097:blood*; D002692:Chlamydia trachomatis / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D012680:Sensitivity and Specificity; D001616:beta-Galactosidase
D018834:Chaperonin 60; D001616:beta-Galactosidase