title: Identification of Suitable Candidate Reference Genes for Gene Expression Analysis by RT-qPCR in Peripheral Blood Mononuclear Cells of CHB Patients.
abstract: BACKGROUND
  Quantitative polymerase chain reaction (qPCR) analysis is a precise and effective method for the study of mRNA expression throughout the field of peripheral blood mononuclear cell (PBMC) research. However, the use of suitable reference genes for data normalization is critical to obtain meaningful and reproducible results. The present study aimed to identify the greatest reference genes for further research in PBMC of Chronic Hepatitis B (CHB) patients.
  
  
  METHODS
  We assessed the expression stability of four commonly used reference genes (beta actin, beta-tubulin, 18S rRNA, GAPDH) in PBMC of CHB patients. Then we employed geNorm, BestKeeper, and Normfinder to evaluate the expression stability of these reference genes.
  
  
  RESULTS
  All four genes displayed no significant differences between patient and control groups except beta actin and thus beta actin should not be used as a normalizing gene in a discussed experimental setup. GAPDH and beta-tubulin composed the best pair of reference genes for normalization purposes in future studies of gene expression in PBMC of CHB patients according to three algorithms.
  
  
  CONCLUSIONS
  GAPDH and beta-tubulin were the best combination of two reference genes in this study for RT-qPCR analysis.
authors.forename: Hao
authors.initials: H
authors.lastname: Zhang
authors.forename: Zi-shu
authors.initials: ZS
authors.lastname: Guan
authors.forename: Shi-he
authors.initials: SH
authors.lastname: Guan
authors.forename: Kai
authors.initials: K
authors.lastname: Yang
authors.forename: Ying
authors.initials: Y
authors.lastname: Pan
authors.forename: Yuan-yuan
authors.initials: YY
authors.lastname: Wu
authors.forename: Ai-hua
authors.initials: AH
authors.lastname: Wang
authors.forename: Bei-bei
authors.initials: BB
authors.lastname: Sun
authors.forename: Jie
authors.initials: J
authors.lastname: Hou
authors.forename: Xuan-xuan
authors.initials: XX
authors.lastname: Mu
authors.forename: Yu-feng
authors.initials: YF
authors.lastname: Gao
authors.forename: Wei-sheng
authors.initials: WS
authors.lastname: Cheng
mesh_terms: D000199:Actins / Q000235:genetics; D000465:Algorithms; D002138:Calibration; D016022:Case-Control Studies; D020869:Gene Expression Profiling / Q000379:methods / Q000592:standards*; D005819:Genetic Markers; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics*; D019694:Hepatitis B, Chronic / Q000097:blood / Q000175:diagnosis / Q000235:genetics*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000737:chemistry* / Q000821:virology; D011237:Predictive Value of Tests; D012333:RNA, Messenger / Q000097:blood / Q000235:genetics*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000592:standards*; D012015:Reference Standards; D014404:Tubulin / Q000235:genetics*
chemical_list: D000199:Actins; D005819:Genetic Markers; D012333:RNA, Messenger; D012336:RNA, Ribosomal, 16S; D014404:Tubulin; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases