Detector sample diluent

All plasma samples were first thawed and centrifuged at 10.000g, +4°C for 10 minutes to remove debris. Because bats can harbor many viruses, supernatants were treated in a P2 laboratory for viral inactivation using a standard solvent/detergent protocol used for human blood plasma products (34 (link), 35 (link)) and described in (36 (link)) and in (32 (link)). Briefly, samples were treated with Tri-n-Butyl Phosphate (TnBP) 0.3% (v/v) and Triton X100 (TX100) 1% (v/v) for 2 hours at room temperature. After treatment, TnBP was removed by passing the samples through a C18 column (Discovery DSC-18 SPE from Supelco). For digital ELISA assays, inactivated samples and stimulated cell supernatants were diluted in the Detector/Sample Diluent (Quanterix) added with NP40 0,5% (v/v). They were then incubated for one hour at room temperature before analysis. Global dilution factor was generally 1/6 for plasma samples and 1/3 for stimulated cell supernatants depending on the amount of material available and to allow the optimal protein detection.

All serum and plasma samples were thawed and centrifugated at 10,000g, +4°C for 10 min. Supernatants were diluted in the Detector / Sample Diluent (Quanterix) for pan-IFNα or IFNα2 quantification, then incubated one hour at room temperature before analysis. Biological samples were diluted from 1/3 to 1/300 depending on the amount of material available and to avoid saturation. NP40 was added in the Detector / Sample Diluent used for cHCV, dengue or viral CNS infection samples at a final concentration of 0.5% (v/v) to inactivate viruses.