Reproducible gene expression measurement among multiple laboratories obtained in a blinded study using standardized RT (StaRT)-PCR.
BACKGROUND
A method that provides standardized data and is relatively inexpensive and capable of high throughput is a prerequisite to the development of a meaningful gene expression database suitable for conducting multi-institutional clinical studies based on expression measurement. Standardized RT (StaRT)-PCR has all these characteristics. In addition, the method must be reproducible. StaRT-PCR has high intralaboratory reproducibility. The purpose of this study is to determine whether StaRT-PCR provides similar interlaboratory reproducibility.


RESULTS
In a blinded interlaboratory study, expression of ten genes was measured by StaRT-PCR in a complementary DNA sample provided to each of four laboratories. The average coefficient of variation for interlaboratory comparison of the nine quantifiable genes was 0.48. In all laboratories, expression of one of the genes was too low to be measured.


CONCLUSIONS
Because StaRT-PCR data are standardized and numerical and the method is reproducible among multiple laboratories, it will allow development of a meaningful gene expression database.
Crawford
E L
EL
Department of Medicine, Medical College of Ohio, Toledo, OH 43699, USA.
Peters
G J
GJ
Noordhuis
P
P
Rots
M G
MG
Vondracek
M
M
Grafström
R C
RC
Lieuallen
K
K
Lennon
G
G
Zahorchak
R J
RJ
Georgeson
M J
MJ
Wali
A
A
Lechner
J F
JF
Fan
P S
PS
Kahaleh
M B
MB
Khuder
S A
SA
Warner
K A
KA
Weaver
D A
DA
Willey
J C
JC
D001667:Binding, Competitive / Q000235:genetics; D002460:Cell Line; D018076:DNA, Complementary / Q000235:genetics; D030541:Databases, Genetic; D004311:Double-Blind Method; D015870:Gene Expression; D020869:Gene Expression Profiling / Q000145:classification / Q000592:standards* / Q000706:statistics & numerical data; D006801:Humans; D008168:Lung / Q000737:chemistry / Q000166:cytology / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000592:standards* / Q000706:statistics & numerical data; D013698:Templates, Genetic; D009626:Terminology as Topic
D018076:DNA, Complementary; D012333:RNA, Messenger