TYPING OF EUROPEAN BLUETONGUE VIRUS SEROTYPES 1, 6, AND 8 BY REAL-TIME RT- PCR

A novel real-time RT-PCR (rRT-PCR) for the detection and typing of bluetongue virus (BTV) in EDTA treated blood samples taken from BTV infected animals was described. This rRT-PCR was based on BTV Seq-2 target gene encoding the highly variable outer shell protein VP2. The applied PCR was accurate, specific, and reliable technique for the detection of a specific sequence for a BTV type and endogenous internal positive control (IPC) in the same well. Using this technique, it was possible to identify the European BTV serotypes I, 6, and 8 in archival blood samples supplied during 2008-2010 for the purpose of the ring trial for BTV genome and antibody detection. Moreover, it was shown that all archive BTV positive blood samples taken from seropositive cattle imported to Poland from Germany were positive for BTV8. This method was much faster (approximately 4 h) and more precise than conventional serological typing methods. In addition, the used thermal cycler allows the use of 96-well plate formats, which further increases the capacity and speed of the analysis. Therefore, it seems to be a valuable complementing tool for a routine diagnosis of BTV infection.