Peptidylpropyl isomerase B (PPIB): a suitable reference gene for mRNA quantification in peripheral whole blood.
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.
Department of Human Genetics, BioMérieux, 69280 Marcy-l'Etoile, France. alexandre.pachot@eu.biomerieux.com
Alexandre
A
Pachot
Jean-Luc
JL
Blond
Bruno
B
Mougin
Pierre
P
Miossec
D019985:Benchmarking / Q000379:methods; D001774:Blood Chemical Analysis / Q000379:methods; D021983:Cyclophilins / Q000032:analysis* / Q000097:blood* / Q000235:genetics* / Q000592:standards; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D019696:Peptidylprolyl Isomerase; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012333:RNA, Messenger / Q000097:blood* / Q000592:standards; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D012333:RNA, Messenger; C081786:cyclophilin B; D021983:Cyclophilins; D019696:Peptidylprolyl Isomerase