Homogenization and Viral Genome Analysis

For the homogenization of organ samples in a serum-free medium, the TissueLyzer II tissue homogenizer (Qiagen, Hilden, Germany) was used. Subsequently, the DNA of all samples taken during the study was extracted using the KingFisher Flex System (Thermo Scientific, Darmstadt, Germany) and the NucleoMag Vet kit (Macherey-Nage, Düren, Germany) according to the manufacturer’s instructions. During DNA extraction, an internal control-DNA (IC-2 DNA) was added to the samples to control successful DNA extraction and inhibition-free amplification [44 (link),45 (link)]. The analysis of the viral genome load in the samples was performed using the PerfeCTa qPCR ToughMix (Quanta BioSciences, Gaithersburg, MD, USA) and the already described pan capripox real-time qPCR of Bowden et al. [6 (link)] with a modified probe [46 (link)].