Nucleospin blood and nucleospin tissue kits

The extraction of viral DNA from blood samples, milk, skin scarifications, skin biopsies, internal organs, and nasal swabs of animal origin, as well as swabs and samples from the environment and insects, was performed at VSI by using a commercial MagVet Universal DNA/RNA kit (LSI, Germany) and Bioextract Superball (Biosellal, Dardilly, France) on Kingfisher mL and Kingfisher Flex (Thermo Fisher Scientific, Vantaa, Finland) according to the manufacturer’s instructions. The extracted DNA was stored at temperatures below −16 °C until the start of the testing.
Viral DNA was extracted at Sciensano using the NucleoSpin Blood and NucleoSpin tissue kits (Macherey-Nagel, Düren, Germany) as previously described [21 (link)].

Viral DNA from blood (200 µL), tissue and organ samples (25 mg) was extracted using the NucleoSpin Blood and NucleoSpin tissue kits (Macherey-Nagel, Düren, Germany), as described by the manufacturer with the exception of: (a) the incubation time for lysis of blood samples was prolonged to 1 h; (b) an external control (EC) was added to B3 buffer before extraction [31 (link)]; (c) tissue/organ samples were homogenized using a TissueLyser (Qiagen, Gaithersburg, MD, USA) in the presence of buffer T1 and proteinase K before the overnight incubation at 56 °C. Viral DNA from swabs was extracted by placing them into 1 mL PBS for 15 min at room temperature. Subsequently, the swabs were vortexed for 1 min and the DNA was extracted using the NucleoSpin Blood kit, as described above.