Since 2006, the occurrence of bluetongue in Northwest- and Central Europe has lead to extensive financial losses. For first-line laboratory diagnosis, reverse transcription real-time PCR (RT-qPCR) has been used increasingly, necessitating careful evaluation of all applied methods by the performing laboratory. In the course of an internal validation, four commercially available BTV RT-qPCR kits and two published reference methods were compared. Clear differences regarding reaction kinetics, analytical as well as diagnostic sensitivity and specificity were observed. Furthermore, test kits were not equally suitable for testing samples from a selection of BTV-susceptible host species. Only two commercial kits were in all examined parameters equal to, or superior than, the more demanding reference methods and thus represent a possible alternative to using published methods for BTV RT-qPCR screening.