Purity for clarity: the need for purification of tumor cells in DNA microarray studies.
It is now well established that gene expression profiling using DNA microarrays can provide novel information about various types of hematological malignancies, which may lead to identification of novel diagnostic markers. However, to successfully use microarrays for this purpose, the quality and reproducibility of the procedure need to be guaranteed. The quality of microarray analyses may be severely reduced, if variable frequencies of nontarget cells are present in the starting material. To systematically investigate the influence of different types of impurity, we determined gene expression profiles of leukemic samples containing different percentages of nonleukemic leukocytes. Furthermore, we used computer simulations to study the effect of different kinds of impurity as an alternative to conducting hundreds of microarray experiments on samples with various levels of purity. As expected, the percentage of erroneously identified genes rose with the increase of contaminating nontarget cells in the samples. The simulations demonstrated that a tumor load of less than 75% can lead to up to 25% erroneously identified genes. A tumor load of at least 90% leads to identification of at most 5% false-positive genes. We therefore propose that in order to draw well-founded conclusions, the percentage of target cells in microarray experiment samples should be at least 90%.
de Ridder
D
D
Department of Immunology, Erasmus MC, University Medical Center Rotterdam, Rotterdam, The Netherlands.
van der Linden
C E
CE
Schonewille
T
T
Dik
W A
WA
Reinders
M J T
MJ
van Dongen
J J M
JJ
Staal
F J T
FJ
D002469:Cell Separation / Q000379:methods*; D003198:Computer Simulation; D004273:DNA, Neoplasm / Q000302:isolation & purification*; D015973:Gene Expression Regulation, Leukemic; D017408:Guidelines as Topic; D006801:Humans; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000235:genetics*; D015459:Leukemia-Lymphoma, Adult T-Cell / Q000235:genetics*; D007963:Leukocytes, Mononuclear / Q000166:cytology; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods* / Q000592:standards; D015203:Reproducibility of Results
D004273:DNA, Neoplasm