Supplementary Online‑only Supplementary File 1 Questionnaire used during the workshop. The domain (D) to which each question belonged to in written beside each question. These domains were not mentioned in the questionnaires given to participants. Domain 1 (D1): Antigen‑antibody interactions and laboratory techniques other than immunofluorescence Domain 2 (D2): Immunofluorescence Domain 3 (D3): Clinical relevance of autoantibodies 1. In a precipitation reaction (D1) • Ag in solid phase, Ab in liquid phase • Ag in liquid phase, Ab in solid phase • Both Ag and Ab in solid phase • Both Ag and Ab in liquid phase. 2. Nephelometry detects (D1) • Scattered light • Transmitted light • Both •. Neither 3. Antinuclear antibody slides are made from (D2) • Hep2 cells • Jurkat cells • Power cells • Peripheral blood lymphocytes. 4. Antineutrophil cytoplasmic antibody stands for (D2) • Anti‑neutrophilic cytosolic antigen • Anti‑neuronal cytoplasmic antibody • Anti‑neutrophil cytoplasmic antibody • Anti‑netosis cellular agglutinant. 5. Antinuclear antibody test utilizes (D2) • Direct immunofluorescence • Indirect immunofluorescence • Direct immunofixation • Indirect immunofixation. 6. A homogenous antinuclear antibody pattern reflects (D2) • Staining of the entire nucleoplasm • Staining of the entire nuclear membrane • Staining of the chromatin in the dividing cells • Staining of the nucleoli. 7. A patient with SLE can have (D3) • Homogenous pattern • Coarse speckled pattern • Either homogenous or coarse speckled pattern • Nucleolar pattern. 8. Anticentromere antibody predicts (D3) • Higher risk of scleroderma renal crisis • Higher incidence of diffuse skin involvement • Higher risk of developing scleroderma in a healthy individual • Higher risk of mortality in limited scleroderma. 9. A patient with inflammatory myositis is likely to have (D3) • Speckled antinuclear antibody • Cytoplasmic staining (antinuclear antibody negative) • Either speckled antinuclear antibody or cytoplasmic staining • Nuclear envelope staining. 10. In formalin‑fixed neutrophils (D2) • both c‑antineutrophil cytoplasmic antibody and p‑antineutrophil cytoplasmic antibody show cytoplasmic fluorescence • both c‑antineutrophil cytoplasmic antibody and p‑antineutrophil cytoplasmic antibody show perinuclear fluorescence • c‑antineutrophil cytoplasmic antibody shows cytoplasmic fluorescence, but p‑antineutrophil cytoplasmic antibody shows perinuclear fluorescence • c‑antineutrophil cytoplasmic antibody shows perinuclear fluorescence, but p‑antineutrophil cytoplasmic antibody shows cytoplasmic fluorescence. 11. What antibodies cannot be made out rat liver kidney slides (D2) • Anti‑liver‑kidney microsomal • Anti‑liver cytosolic antibody • Anti‑mitochondrial antibody • Anti‑melanocyte antibody. 12. Immunofluorescence is the phenomenon of (D2) • Emission of light in ultraviolet spectrum by excited fluorochrome • Emission of light inversely proportional to the degree of staining by excited fluorochrome • Emission of light in visible spectrum by excited fluorochrome • Emission of transmitted light through dividing cells seen under fluorescence microscope. 13. Antinuclear antibody by enzyme‑linked immunosorbent assay (D3) • Has low sensitivity • Should be the standard of care • Should not be performed • Has high specificity. 14. Antiphospholipid antibodies (D3) • Should be repeated at least after 24 weeks and before 5 years • Should be repeated at least after 10 weeks and before 8 years • Should be repeated at least after 12 weeks and before 5 years • Should be repeated at least after 12 months and before 5 years. 15. A standard curve (D1) • Requires patient samples of graded concentrations in each well • Requires standard samples of similar concentrations in each well • Requires standard samples of graded concentrations in each well • Requires standard samples of identical concentration in each well. 16. Which is not a myositis‑specific antibody (D3) • Anti‑U1RNP antibody • Anti‑SRP antibody • Anti‑MDA‑5 antibody • Anti‑Mi2 antibody. 17. Anti‑RNP antibodies (D3) • Are specific for mixed connective tissue disease • Are subject to epitope spreading • Are not valid makers of disease activity in mixed connective tissue disease • Are useful to differentiate lupus from myositis. 18. The Crithidia luciliae assay looks at (D2) • Anti‑DNA antibodies • Anti‑dsDNA antibodies • Anti‑ssDNA antibodies • Low affinity anti‑dsDNA and anti‑histone antibodies. 19. Antibodies to cardiolipins in lupus (D3) • Predict cognitive dysfunction • Predict transverse myelitis • Predict membranous nephropathy • Predict aseptic meningitis. 20. Anti‑RNA polymerase III antibody in scleroderma (D3) • Predicts limited cutaneous involvement • Predicts scleroderma renal crisis • Predicts increased risk of myocardial infarction • Predicts higher risk of cutaneous ulcers. 21. Avidity of antibody refers to (D1) • Sum total of all the affinities to all the antibodies and antigenic determinants • Strength of binding between a single antibody and its antigenic determinant site • Interaction between an epitope and paratope • Flexibility of antibody at its hinge region. 22. Antibodies bind to antigen through (D1) • Covalent bond • Noncovalent bond. 23. Prozone phenomenon refers to (D1) • Excess of antigen • Excess of antibody • Excess of either • Absence of salts in the precipitation reaction. 24. Agglutination reactions usually refers to all EXCEPT (D1) • Interaction between solid‑phase antigen and liquid‑phase antibody • Plant proteins, viruses, salts of heavy metals, and basic proteins may cause agglutination • IgG immunoglobulins are more likely to cause agglutination as compared to IgM immunoglobulins. 25. Enzyme‑linked immunosorbent assay plate is made up of (D1) • Polystyrene • Plastic • Glass •. Rubber