Transcription of reference genes used for quantitative RT-PCR in Atlantic salmon is affected by viral infection
1 Section for immunoprophylaxis, National Veterinary Institute, P.O. Box 750 Sentrum, 0106 Oslo, Norway
2 Department of Basic Sciences and Aquatic Medicine, Norwegian School of Veterinary Science, P.O. Box 8146 Dep, 0033 Oslo, Norway
3 Norwegian College of Fishery Science, University of Tromsø, 9037 Tromsø, Norway
4 Department of Food Safety and Infection Biology, Norwegian School of Veterinary Science, P.O. Box 8146 Dep, 0033 Oslo, Norway
5 Intervet Norbio AS, Thormøhlensgate 55, 5008 Bergen, Norway
Veterinary Research 2011, 42:8 doi:10.1186/1297-9716-42-8Published: 18 January 2011
Relative quantification using RT-qPCR is a widely used method for transcription profiling. Transcript levels of target genes in fish after experimental infection is often reported without documentation of stably transcribed reference genes. We present results demonstrating that transcription of typically used reference genes in Atlantic salmon is not stable during experimental infection with salmon pancreas disease virus (SPDV). Transcript levels 0 to 6 weeks after challenge revealed statistically significant changes between time-points that corresponded with a peak in viral load 3 weeks after challenge. The results emphasize the need for thorough method validation prior to transcriptional studies during viral infections.