The objective of this study was to analyze apparently discrepant results that arose during the use of an indirect immunofluorescence (IIF) assay using transfected HEp-2 cells to detect anti-SS-A/Ro autoantibodies in human sera. Fourteen sera that had SS-A/Ro antibodies as detected on this commercial substrate, but did not have antibodies to SS-A/Ro as determined by double immunodiffusion (ID) or enzyme-linked immunosorbent assay (ELISA), were studied by immunoprecipitation (IP) of radiolabeled cell extracts and full-length recombinant SS-A/Ro. A multi-antigen strip immunoblotting (IB) assay containing both the 52- and 60-kDa antigens was included in the analysis. We confirmed that 12 of 14 of the sera under study had antibodies to SS-A/Ro protein antigens as determined by at least one other immunoassay. One serum had antibodies to hyRNA but no detectable reactivity with the 52- or 60-kDa antigens. One serum remained negative in all assays for SS-A/Ro autoantibodies.
Copyright 2002 Wiley-Liss, Inc.