Laboratory diagnosis
For qPCR at the regional laboratory (Landeslabor Berlin-Brandenburg,
Frankfurt/Oder, Germany, LLBB), the commercial RealPCR ASFV DNA Test
(IDEXX, Westbrook, USA) was used following the manufacturer’s
instructions. For confirmatory purposes, the OIE-listed protocol by King
et al. (2003) was used with slight modifications. At the German national
reference laboratory (NRL at the FLI), qPCRs were performed according to
the protocols published by King et al. (2003) and Tignon et al. (2011).
Both assays were used with adapted internal control systems. In
addition, a commercial qPCR kit was employed (virotype ASFV, Indical
Bioscience, Leipzig, Germany) according to the manufacturer´s
instructions. To isolate the causative ASFV in serum and tissue samples,
a haemadsorption test (HAT) was performed using PBMC-derived macrophages
according to slightly modified standard procedures (Carrascosa, Bustos,
& de Leon, 2011).