Peptidylpropyl isomerase B (PPIB): a suitable reference gene for mRNA quantification in peripheral whole blood

J Biotechnol. 2004 Oct 19;114(1-2):121-4. doi: 10.1016/j.jbiotec.2004.07.001.

Abstract

Quantitative real-time RT-PCR is a very powerful technique for measuring gene expression at the mRNA level. In order to compare mRNA expression in different experimental or clinical conditions, expression of a target gene has to be normalized to an appropriate internal standard, which is generally a housekeeping gene. In our study, we have tested several housekeeping genes in peripheral whole blood of healthy volunteers and patients suffering from inflammatory diseases. A first analysis of 91 samples illustrated that the mRNA expression of peptidylpropyl isomerase B (PPIB) encoding for cyclophilin B protein, is more stable than beta actin and glyceraldehyde-3-phosphate dehydrogenase, which are both commonly selected as internal standard. Among the three genes tested, beta actin displayed the highest inter-sample variation of expression. The constancy of PPIB mRNA expression was further confirmed by 214 additional samples. In conclusion, we showed that PPIB, in contrast to beta actin and glyceraldehyde-3-phosphate dehydrogenase, is a suitable housekeeping gene in human peripheral blood.

Publication types

  • Comparative Study
  • Evaluation Study
  • Validation Study

MeSH terms

  • Benchmarking / methods
  • Blood Chemical Analysis / methods
  • Cyclophilins / analysis*
  • Cyclophilins / blood*
  • Cyclophilins / genetics*
  • Cyclophilins / standards
  • Gene Expression Profiling / methods*
  • Humans
  • Peptidylprolyl Isomerase
  • Polymerase Chain Reaction / methods*
  • Polymerase Chain Reaction / standards
  • RNA, Messenger / blood*
  • RNA, Messenger / standards
  • Reference Standards
  • Reproducibility of Results
  • Sensitivity and Specificity

Substances

  • RNA, Messenger
  • cyclophilin B
  • Cyclophilins
  • Peptidylprolyl Isomerase