SARS-CoV-2 Antibody Detection from Dried Blood Spots
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Corresponding Organization : University of Massachusetts Amherst
Variable analysis
- Heat treatment of sample cards (30 minutes at 56° C)
- SARS-CoV-2 antibody levels
- Protein Saver 903 sample cards
- Punching of a single blood spot per card (0.25-inch diameter)
- Coating of ELISA plate with 1 μg/ml of purified RBD diluted in PBS overnight at 4°C
- Blocking of ELISA plate with Tris-Buffered Saline with 0.1% Tween 20 (TBST) containing 5% non-fat dry milk
- Elution of DBS in 500 μl of TBST overnight at 4°C
- Addition of 50 μl of each sample to the ELISA plate preloaded with 50 μl of TBST containing 2% non-fat dry milk
- Optical density reading at 405 nm
- Inclusion of seven negative controls and one positive control (serum from PCR-confirmed case at 1/100 dilution) per 96-well plate
- Normalization of optical density values to the mean optical density of negative controls daily
- Serum from PCR-confirmed case at 1/100 dilution
- Seven negative controls per 96-well plate
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