Med. Weter. 68 (12), 736-739, 2012

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Niedbalski W.
Validation of universal and serotype-specific real-time RT- PCR assays for the detection of European bluetongue virus serotypes
The aim of this study was to validate real-time RT-PCR (rRT-PCR) assays for the detection and typing of bluetongue virus (BTV) serotypes recently circulating in Europe. The universal rRT-PCR assay (for all BTV genotypes) was based on a highly conserved region in BTV RNA segment 1, and the serotype–specific (BTV typing) rRT-PCR was based on the BTV Seg-2 target gene encoding the highly variable outer shell protein VP2. The rRT-PCR techniques applied here are very fast (approximately 4 h), specific and sensitive for the detection and identification of BTV serotypes. Using the BTV-typing rRT-PCR, it was possible to identify European BTV serotypes 1, 2, 4, 6, 8, 11 and 16 in archival blood samples collected in 2008-2011 for the purpose of a ring trial for BTV genome and antibody detection. This assay may therefore be considered as a valuable tool complementing the routine diagnostic procedure for BTV diagnosis.
Key words: bluetongue virus, detection, typing, real-time RT-PCR