Reference genes evaluated for use in infectious pancreatic necrosis virus real-time RT-qPCR assay applied during different stages of an infection.
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.
Julin
K
K
Nofima Marin, N-9291 Tromsø, Norway. kjersti.julin@nofima.no
Johansen
L-H
LH
Sommer
A-I
AI
D000199:Actins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D018175:Birnaviridae Infections / Q000662:veterinary* / Q000821:virology; D002460:Cell Line; D005393:Fish Diseases / Q000821:virology*; D020869:Gene Expression Profiling / Q000592:standards; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics / Q000378:metabolism; D018061:Infectious pancreatic necrosis virus / Q000235:genetics / Q000302:isolation & purification / Q000472:pathogenicity*; D007668:Kidney / Q000378:metabolism / Q000821:virology; D011708:Pylorus / Q000378:metabolism / Q000821:virology; D012337:RNA, Ribosomal, 18S / Q000235:genetics / Q000378:metabolism; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000592:standards*; D012269:Ribosomal Proteins / Q000235:genetics / Q000378:metabolism; D020317:Salmo salar / Q000821:virology*; D012474:Salmon / Q000821:virology*
D000199:Actins; D012337:RNA, Ribosomal, 18S; D012367:RNA, Viral; D012269:Ribosomal Proteins; C029611:ribosomal protein S20; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases