2.5.3. Plaque Reduction Neutralization Test (PRNT)

The protocol for the Plaque Reduction Neutralization Test (PRNT) was already published [34]. Briefly, serial serum dilutions were incubated with approx. 50 pfu NSDV, before the serum/virus mixture was transferred to prepared SW13 cell monolayers in a 6-well format. After an incubation period, the inocula were removed and an overlay was added to each well. Plates were fixed after 5 days, and a serum dilution was qualified positive if 80% of plaques (PRNT₈₀) were reduced compared to the control plate.