Reactive oxygen species modify human DNA, eliciting a more discriminating antigen for the diagnosis of systemic lupus erythematosus.
During the development of an ELISA to measure anti-DNA antibodies in systemic lupus erythematosus (SLE) sera, native dsDNA was found not to be the most appropriate antigen to use in ELISA assays for differentiating between SLE patients and those with rheumatoid arthritis (RA), a disease also associated with circulating serum anti-DNA antibodies. By modifying the ELISA technique to incorporate human DNA, denatured by reactive oxygen species, to detect anti-DNA antibodies in SLE sera, results consistently showed an increase in antibody binding when compared with the native antigen; no such trend was observed in the comparable group of RA patients. Using this assay serum anti-dsDNA antibody levels were measured in a group of 20 controls, 20 RA patients (10 seropositive and 10 seronegative) and 30 SLE patients (15 with clinically active disease, 15 with inactive disease). A comparison with the standard radioimmunoassay used to measure anti-DNA antibodies for the diagnosis of SLE showed that the ELISA assay using modified DNA performed better than the standard radioimmunoassay offering an improvement in both clinical specificity and sensitivity. The improved method particularly reduced the problem of false-negative results for SLE patients shown clinically to be either mildly active or inactive.
Blount
S
S
Department of Rheumatology, University of Birmingham, England, UK.
Griffiths
H
H
Emery
P
P
Lunec
J
J
D000328:Adult; D000368:Aged; D000918:Antibody Specificity; D000941:Antigens / Q000276:immunology*; D001172:Arthritis, Rheumatoid / Q000175:diagnosis; D001323:Autoantibodies / Q000032:analysis / Q000276:immunology; D004247:DNA / Q000276:immunology* / Q000378:metabolism; D003937:Diagnosis, Differential; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D006801:Humans; D008180:Lupus Erythematosus, Systemic / Q000175:diagnosis* / Q000276:immunology; D008297:Male; D008875:Middle Aged; D009691:Nucleic Acid Denaturation; D010100:Oxygen / Q000378:metabolism*; D011237:Predictive Value of Tests; D011863:Radioimmunoassay
D000941:Antigens; D001323:Autoantibodies; D004247:DNA; D010100:Oxygen