title: Quality-controlled measurement methods for quantification of variations in transcript abundance in whole blood samples from healthy volunteers.
abstract: BACKGROUND
  Transcript abundance (TA) measurement in whole blood frequently is conducted to identify potential biomarkers for disease risk and to predict or monitor drug response. Potential biomarkers discovered in this way must be validated by quantitative technology. In this study we assessed the use of standardized reverse transcription PCR (StaRT-PCR) to validate potential biomarkers discovered through whole blood TA profiling.
  
  
  METHODS
  For each of 15 healthy volunteers, 6 blood samples were obtained, including 3 samples at each of 2 separate visits. Total variation in TA for each gene was partitioned into replicate, sample, visit, study participant, and residual components.
  
  
  RESULTS
  Variation originating from technical processing was <5% of total combined variation and was primarily preanalytical. Interindividual biological sample variation was larger than technical variation. For 12 of 19 tests, the distribution of measured values was gaussian (Shapiro-Wilks test).
  
  
  CONCLUSIONS
  For control or diseased population groups with variation rates as low as those observed in this control group, 17 individuals per group would be required to detect 1 SD change with 80% power with a 2-sided alpha = 0.05 statistical test for mean differences.
authors: Gene Express, Inc., Toledo, OH, USA.
authors: Elizabeth Herness
authors: EH
authors: Peters
authors: Sandra
authors: S
authors: Rojas-Caro
authors: Mitchell G
authors: MG
authors: Brigell
authors: Robert J
authors: RJ
authors: Zahorchak
authors: Shelley Ann
authors: SA
authors: des Etages
authors: Patricia L
authors: PL
authors: Ruppel
authors: Charles R
authors: CR
authors: Knight
authors: Bradley
authors: B
authors: Austermiller
authors: Myrna C
authors: MC
authors: Graham
authors: Steve
authors: S
authors: Wowk
authors: Sean
authors: S
authors: Banks
authors: Lakshmi V
authors: LV
authors: Madabusi
authors: Patrick
authors: P
authors: Turk
authors: Donna
authors: D
authors: Wilder
authors: Carole
authors: C
authors: Kempfer
authors: Terry W
authors: TW
authors: Osborn
authors: James C
authors: JC
authors: Willey
mesh_terms: D015415:Biomarkers / Q000097:blood*; D003627:Data Interpretation, Statistical; D020869:Gene Expression Profiling / Q000592:standards* / Q000706:statistics & numerical data; D014644:Genetic Variation; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000592:standards* / Q000706:statistics & numerical data; D011786:Quality Control; D012016:Reference Values; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D015415:Biomarkers