Reference gene selection for quantitative real-time PCR analysis in virus infected cells: SARS corona virus, Yellow fever virus, Human Herpesvirus-6, Camelpox virus and Cytomegalovirus infections

Virol J. 2005 Feb 10:2:7. doi: 10.1186/1743-422X-2-7.

Abstract

Ten potential reference genes were compared for their use in experiments investigating cellular mRNA expression of virus infected cells. Human cell lines were infected with Cytomegalovirus, Human Herpesvirus-6, Camelpox virus, SARS coronavirus or Yellow fever virus. The expression levels of these genes and the viral replication were determined by real-time PCR. Genes were ranked by the BestKeeper tool, the GeNorm tool and by criteria we reported previously. Ranking lists of the genes tested were tool dependent. However, over all, beta-actin is an unsuitable as reference gene, whereas TATA-Box binding protein and peptidyl-prolyl-isomerase A are stable reference genes for expression studies in virus infected cells.

MeSH terms

  • Cell Line
  • Cytomegalovirus / genetics
  • Cytomegalovirus / isolation & purification*
  • Genes, Viral / genetics*
  • Herpesvirus 6, Human / genetics
  • Herpesvirus 6, Human / isolation & purification*
  • Humans
  • Peptidylprolyl Isomerase / genetics
  • Peptidylprolyl Isomerase / metabolism
  • Polymerase Chain Reaction / methods*
  • Polymerase Chain Reaction / standards
  • Poxviridae / genetics
  • Poxviridae / isolation & purification*
  • Reference Standards
  • Severe acute respiratory syndrome-related coronavirus / genetics
  • Severe acute respiratory syndrome-related coronavirus / isolation & purification*
  • TATA-Box Binding Protein / metabolism
  • Virus Replication
  • Yellow fever virus / genetics
  • Yellow fever virus / isolation & purification*

Substances

  • TATA-Box Binding Protein
  • Peptidylprolyl Isomerase