Nofima Marin, N-9291 Tromsø, Norway.
J Virol Methods. 2009 Dec;162(1-2):30-9. doi: 10.1016/j.jviromet.2009.07.003. Epub 2009 Jul 26.
The stability of 6 reference genes, 18S, beta-actin, RPS20, eEF1alpha, G6PDH and GAPDH, was examined in tissues from Atlantic salmon (Salmo salar) and Chinook salmon embryo cells (CHSE-214). The main objective of this study was to determine the most suitable reference genes for use for the normalisation of data in quantitative real-time RT-qPCR assays conducted on infected tissues. The tissue samples selected for analysis were taken from head kidney and pylorus and collected at different time points during a challenge experiment with infectious pancreatic necrosis virus (IPNV). The stability of some of the reference genes was also studied in infected CHSE-214 cells. The ranking of the genes examined was carried out using the geNorm program. This program determines the most stable genes from a set of genes tested in a given cDNA sample. The stability of the reference genes varied in different tissues and in the cell line at different stages of infection with IPNV. This study demonstrated that tissue-specific combinations of reference genes must be used to normalise real time data for use for the quantitation of IPNV.
6 个参照基因（18S、β-肌动蛋白、RPS20、eEF1alpha、G6PDH 和 GAPDH）在大西洋鲑（Salmo salar）和奇努克鲑鱼胚胎细胞（CHSE-214）组织中的稳定性进行了检测。本研究的主要目的是确定最适合用于感染组织定量实时 RT-qPCR 分析的正常化数据的参照基因。选择用于分析的组织样本取自头肾和幽门，并在传染性胰腺坏死病毒（IPNV）感染实验的不同时间点采集。一些参照基因的稳定性也在感染的 CHSE-214 细胞中进行了研究。使用 geNorm 程序对所检查的基因进行了排名。该程序从给定 cDNA 样本中测试的一组基因中确定最稳定的基因。参照基因的稳定性在不同组织和感染 IPNV 的细胞系的不同阶段有所不同。本研究表明，必须针对特定组织的参照基因组合进行实时数据的正常化，以用于 IPNV 的定量。
