Evaluation of a recombinant 27-kDa outer membrane protein of Coxiella burnetii as an immunodiagnostic reagent.
The 27-kDa outer membrane protein from eight strains of Coxiella burnetii was expressed in the pET-21c protein expression system. Two fusion proteins with molecular masses of 30 and 32 kDa were evident in all eight of the recombinants by SDS-PAGE and immunoblotting. A protein having an approximate size of 30 kDa was purified from the Escherichia coli lysates by one-step affinity purification. The utility of the purified recombinant protein in ELISA was also evaluated by testing its reactivity with human sera and comparing this reactivity with that of Nine Mile phase II antigen. All of the 40 IF-positive serum samples were ELISA-positive for both the Nine Mile phase II and recombinant antigens, and negative serum controls were negative for both antigens. These results suggest that ELISA with the 27-kDa recombinant antigen is a sensitive and specific method for detecting anti-C. burnetii antibodies in human sera.
Department of Veterinary Microbiology, Faculty of Agriculture, Gifu University, Japan.
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GQ
Zhang
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A
Hotta
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T
Ho
T
T
Yamaguchi
H
H
Fukushi
K
K
Hirai
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial / Q000276:immunology; D001425:Bacterial Outer Membrane Proteins / Q000276:immunology*; D016997:Coxiella burnetii / Q000276:immunology* / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005069:Evaluation Studies as Topic; D005260:Female; D006801:Humans; D015151:Immunoblotting; D008297:Male; D051379:Mice; D011778:Q Fever / Q000175:diagnosis*; D011993:Recombinant Fusion Proteins / Q000276:immunology*; D012680:Sensitivity and Specificity
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D011993:Recombinant Fusion Proteins