Serological Assay for CCHFV Antibodies
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Corresponding Organization :
Other organizations : Office of Extramural Research, Karolinska University Hospital, Karolinska Institutet, Public Health Agency of Sweden, University of Giessen
Variable analysis
- Coating of Nunc Maxisorp plates with 200ng/well of mouse monoclonal antibodies against NP (clone 9D5, BEI Resources) or Gc (11E7, BEI resources)
- Whole-CCHFV-virion-specific IgM and IgG responses in serum, quantified by an in-house ELISA
- NP or Gc specific responses, measured by a sandwich ELISA
- Blocking of plates with 5% skim milk in PBS 0.05% Tween
- Application of semi-purified whole virus lysate
- Serum dilution at 1:1000
- Detection of bound antibodies with goat horseradish peroxidase conjugated anti-monkey IgG (Seracare, catalog KPL 074–11-021) at 1:2000
- Development with ABTS solution (Seracare) and stopping the reaction with 5% sodium dodecyl sulfate in water
- Absorbance reading at 405nm on a Synergy HTX (Biotek)
- Subtraction of absorbance values of wells coated with anti-NP or anti-Gc but receiving no CCHFV lysate from wells receiving lysate
- Measurements performed in duplicate
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