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Antigen capture ELISA for the heat shock protein (hsp60) of Chlamydia trachomatis
1Department of Genito-urinary Medicine and Communicable Diseases, Imperial College School of Medicine at St Mary's, London.
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.