Quality-controlled measurement methods for quantification of variations in transcript abundance in whole blood samples from healthy volunteers.
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.
Peters
Elizabeth Herness
EH
Gene Express, Inc., Toledo, OH, USA.
Rojas-Caro
Sandra
S
Brigell
Mitchell G
MG
Zahorchak
Robert J
RJ
des Etages
Shelley Ann
SA
Ruppel
Patricia L
PL
Knight
Charles R
CR
Austermiller
Bradley
B
Graham
Myrna C
MC
Wowk
Steve
S
Banks
Sean
S
Madabusi
Lakshmi V
LV
Turk
Patrick
P
Wilder
Donna
D
Kempfer
Carole
C
Osborn
Terry W
TW
Willey
James C
JC
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
D015415:Biomarkers