Sensitivity of the HEp-2000 substrate for the detection of anti-SSA/Ro60 antibodies

Anti-SSA/Ro antibodies are the most prevalent type of antinuclear antibody (ANA). Anti-SSA/ Ro-positive sera may recognise two proteins: a 52 kDa (Ro52) and a 60 kDa (Ro60) subunit. We studied the sensitivity for Ro60 detection using the HEp-2000 substrate, which consists of HEp-2 cells transfected...

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Bibliographic Details
Published in:Clinical rheumatology 2000-01, Vol.19 (4), p.291-295
Main Authors: Peene, I, Van Ael, W, Vandenbossche, M, Vervaet, T, Veys, E, De Keyser, F
Format: Article
Language:English
Subjects:
Adult
Antibodies, Antinuclear - blood
Antigens, Nuclear
Autoantigens - immunology
DNA-Binding Proteins - immunology
Female
Fluorescent Antibody Technique, Indirect
Humans
Immunoassay
Immunodiffusion
Infant, Newborn
Lupus Erythematosus, Systemic - immunology
Male
Nuclear Proteins - immunology
Ribonucleoproteins - immunology
Sensitivity and Specificity
Staining and Labeling
Transfection
Tumor Cells, Cultured
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Summary:Anti-SSA/Ro antibodies are the most prevalent type of antinuclear antibody (ANA). Anti-SSA/ Ro-positive sera may recognise two proteins: a 52 kDa (Ro52) and a 60 kDa (Ro60) subunit. We studied the sensitivity for Ro60 detection using the HEp-2000 substrate, which consists of HEp-2 cells transfected with Ro60 cDNA in an anti-SSA/Ro-positive population consecutively identified by double immunodiffusion (DID) with thymus/spleen nuclear extract and line immunoassay (LIA) with recombinant Ro52 and Ro60. One hundred and twenty-seven consecutive anti-SSA/Ro-positive sera defined by DID with thymus/spleen nuclear extract and LIA using recombinant Ro52 and Ro60 were analysed on HEp-2000 and DID with natural Ro60. Of these, 91 were anti-Ro60 positive on LIA and/ or DID with natural Ro60. The HEp-2000 substrate detected 70/91 (sensitivity 77%) and correlated strongest with DID. Most of the missed anti-Ro60-positive sera had high ANA intensity. The substrate did not detect monospecific anti-Ro52 antibodies (sensitivity 9.7%; 3/31). HEp-2000 substrate can therefore be considered a reliable, simple and alternative method for DID in the detection of anti-Ro60 reactivity. Special follow-up should be given to sera with strong ANA patterns in which the SSA/Ro60 staining pattern may be hidden.
ISSN:0770-3198
1434-9949
DOI:10.1007/s100670070048