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.