Nucleic acid extraction and real-time PCR
Viral nucleic acids were extracted using the QIAamp® RNA Viral Mini Kit (Qiagen) according to the manufacturer’s instructions. qPCR was conducted according to the protocol published by King et al. (2003) with slight modifications (addition of a heterologous internal control DNA), or with the commercial qPCR kits virotype ASFV or virotype ASFV 2.0 (Indical Bioscience) according to the manufacturer’s instructions. All qPCR runs were performed on C1000™ thermal cyclers with the CFX96™ Real-Time System (Biorad). Results were recorded as quantification cycle (cq) values.