Use of a recombinant Coccidioides immitis complement fixation antigen-chitinase in conventional serological assays.
The coccidioidal complement fixation (CF) antigen has been cloned previously, and the fusion protein has been expressed in Escherichia coli. The recombinant CF (rCF) antigen was affinity purified by adsorption-desorption to chitin, and its reactivity was studied by using sera containing coccidioidal antibodies. The affinity-purified rCF antigen formed a line of identity with an immunodiffusion (ID) CF reference antigen (coccidioidin) derived from mycelial-phase Coccidioides immitis and was reactive with human, canine, and equine sera containing coccidioidal antibody. The affinity-purified rCF antigen yielded no detectable reaction with Blastomyces of Histoplasma antiserum by ID. The affinity-purified rCF antigen fixed complement with positive human sera and, even when used at lower concentrations, yielded titers comparable to those obtained with the coccidioidin. The reactivity of the affinity-purified rCF antigen was further evaluated by enzyme immunoassay, in which it manifested good sensitivity (96.9%) and specificity (100%) when evaluated with 43 human patients' sera. Thus, the affinity-purified rCF antigen has yielded reactions comparable to those of crude coccidioidal antigens in conventional CF, IDCF, and enzyme immunoassay.
Johnson
S M
SM
Department of Medical Microbiology and Immunology, School of Medicine, University of California, Davis 95616, USA.
Zimmermann
C R
CR
Pappagianis
D
D
D000818:Animals; D000908:Antibodies, Fungal / Q000097:blood; D000946:Antigens, Fungal / Q000235:genetics; D002688:Chitinases / Q000235:genetics / Q000276:immunology*; D003045:Coccidioides / Q000201:enzymology / Q000235:genetics / Q000276:immunology*; D003047:Coccidioidomycosis / Q000175:diagnosis / Q000276:immunology; D003168:Complement Fixation Tests / Q000379:methods* / Q000706:statistics & numerical data; D004285:Dogs; D005069:Evaluation Studies as Topic; D006736:Horses; D006801:Humans; D005779:Immunodiffusion / Q000379:methods / Q000706:statistics & numerical data; D007124:Immunoenzyme Techniques / Q000706:statistics & numerical data; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D012680:Sensitivity and Specificity
D000908:Antibodies, Fungal; D000946:Antigens, Fungal; D011994:Recombinant Proteins; D002688:Chitinases