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Abstract 29

Source: www.aacc.org
Topic: Systemic Connective Tissue
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Sort Desciption: Evaluation of an automated multiplexed immuno-blot assay for the detection of antigen specificity of anti-nuclear antibodies in connective tissue diseases. ...

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Evaluation of an automated multiplexed immuno-blot assay for the detection of antigen specificity of anti-nuclear antibodies in connective tissue diseases. Gupta M., Means C. The Cleveland Clinic Foundation, Cleveland, OH. Systemic connective tissue diseases (CTDs) are characterized with the presence of autoantibodies directed against various intracellular antigens. Distinct autoantibody profiles are found in different diseases, hence, determination of antibody specificity is desirable in establishing the correct diagnosis. Currently, individual disease specific anti-nuclear antibodies (ANAs) are detected by immunodiffusion (ID) or by enzyme immunoassays (EIAs), both requiring several individual assays to detect antigen profile, which, is time consuming and expensive. We evaluated a multi-parameter immuno-blot assay (IB) (Innogenetics, Inc. N.Y.) using, mostly recombinant and some natural antigens allowing simultaneous detection of all clinically relevant disease specific ANAs in a single reaction. A total of 9 antigens and 13 determinants are detected in one reaction. These included: Sm (B&D), RNP(70-k, A &C), SS-A/Ro(52 &60), SS-B/La, Cenp-B (centromere), Topo-I/Scl-70, Jo-1/Histidyl tRNA synthetase, ribosomal RNP and Histones. The test is fast (3 hours), simple and semi-automated. A total of 58 samples from patients with known or suspected connective tissue diseases samples, were analyzed with IB and compared with ID for Sm, SSA, SSB and RNP only. These included, SLE/DLE (n=18), Sjogren syndrome (n=11), other CTDs (RA=3; Raynaud =5; CTD=6; APS=2) or no specific CTD pathology (n=13). In addition, 68 control samples (no-CTD) received in the lab from hospital (ICU) were screened for ANA by EIA and seven (10%) were positive. Only two of these were positive with IB (1=SSA only and 1= SSA and SSB). Among the 45 patients with CTDs, the antibody specificities as detected by ID and IB assays were as listed below. Specificity ...

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