The detection of anti-Ro/SS-A and anti-La/SS-B antibodies. A comparison of counterimmunoelectrophoresis with immunoblot, ELISA, and RNA-precipitation assays.
The presence in serum of anti-Ro/SS-A and/or anti-La/SS-B autoantibodies is a characteristic of autoimmune diseases such as Sjögren's syndrome and systemic lupus erythematosus. To evaluate different assays currently available for the detection of these antibodies 50 sera were tested using the four different assay methods: counterimmunoelectrophoresis (CIE), RNA precipitation assay, immunoblotting technique and ELISA. The RNA-precipitation assay showed the highest sensitivity and specificity. The CIE for the detection of anti-Ro/SS-A antibodies gave comparable results whereas the Ro/SS-A ELISA and Ro/SS-A or HeLa immunoblot showed lower sensitivities (96% and 80% respectively). Sensitivity was even lower (66%) when only reactivity towards the 60 kDa Ro/SS-A protein was considered. The ELISA for the detection of anti-La/SS-B antibodies showed a sensitivity of 98%, the immunoblotting technique of 86% and the CIE only 67%. The high sensitivity of the La/SS-B ELISA went together with a low specificity of 14%. We conclude from these data that for the detection of anti-Ro/SS-A and anti-La/SS-B antibodies the RNA precipitation assay shows the highest sensitivity and highest specificity. For routine screening purposes the CIE is the most convenient and reliable assay to detect anti-Ro/SS-A antibodies. For the detection of anti-La/SS-B antibodies the immunoblot corresponds most closely to the RNA precipitation assay.
Central Laboratory of The Netherlands Red Cross Blood Transfusion Service, Amsterdam.
J F
JF
Meilof
I
I
Bantjes
J
J
De Jong
A P
AP
Van Dam
R J
RJ
Smeenk
D000974:Antibodies, Antinuclear / Q000097:blood*; D003378:Counterimmunoelectrophoresis / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting / Q000379:methods*; D011233:Precipitin Tests; D012313:RNA; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000974:Antibodies, Antinuclear; C035356:SS-A antibodies; C035357:SS-B antibodies; D012313:RNA