A novel approach to the laboratory diagnosis of Chlamydia trachomatis infections using monoclonal anti-idiotypic antibodies.
We have developed a novel enzyme immunoassay (EIA) for the specific detection of Chlamydia trachomatis utilizing a monoclonal anti-idiotypic antibody to an antibody directed against a chlamydia specific epitope on 60 kDa heat-shock protein (HSP60). The basis of the assay is the inhibition of the binding of idiotype to anti-idiotype by antigen present in test samples. Two configurations of the assay were developed: a blocking EIA and a competition EIA. Greater sensitivity was observed using the competition EIA, with the assay detecting purified recombinant HSP60 and purified chlamydia in a concentration-dependent manner from 0.01 to 10 micrograms protein and from 0.5 to 12 micrograms total protein, respectively. The assay is highly specific and offers several potential advantages over currently available EIAs for the detection of this pathogen.
Laferrière
C
C
National Laboratory for Immunology, Laboratory Centre for Disease Control, Ottawa, Ontario, Canada.
Peeling
R W
RW
Tackaberry
E S
ES
Hamel
J
J
Dillon
J A
JA
Brodeur
B R
BR
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000276:immunology*; D000907:Antibodies, Bacterial / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000942:Antigens, Bacterial / Q000276:immunology; D001667:Binding, Competitive / Q000276:immunology; D018834:Chaperonin 60; D002690:Chlamydia Infections / Q000175:diagnosis*; D002692:Chlamydia trachomatis / Q000276:immunology*; D006360:Heat-Shock Proteins / Q000032:analysis / Q000235:genetics; D006801:Humans; D007124:Immunoenzyme Techniques; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D007130:Immunoglobulin Idiotypes / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins
D000888:Antibodies, Anti-Idiotypic; D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D018834:Chaperonin 60; D006360:Heat-Shock Proteins; D007140:Immunoglobulin Fab Fragments; D007130:Immunoglobulin Idiotypes; D011993:Recombinant Fusion Proteins