Quality assessment of microarray data in a multicenter study

Kellie J. Archer, Valeria Mas, Thomas R. O'Brien, Ruth Pfeiffer, Nicole L. Lum, Robert A. Fisher

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Issues implicit in a multicenter microarray study are protocol standardization and monitoring center adherence to established protocols. This study explored the effects of submitting center and sample preservation method on the quality of isolated RNA. In addition, the effects of sample preservation method and laboratory on microarray quality were also examined. Herein we evaluated the contribution of specific technical factors [center, laboratory, and preservation method (frozen/RNAlater)] on quality of isolated RNA, cRNA synthesis products, and reproducibility of gene expression microarray data for independent biologic samples collected in a multicenter microarray study. The Kruskal-Wallis test was used to test for differences owing to submitting center on isolated RNA quality. Mixed effects analysis of variance was used in assessing the impact of laboratory and preservation method on gene expression values for the 12 samples hybridized at 2 independent laboratories (24 GeneChips). One center was found to be in violation of the tissue handling protocol. No significant effect was noted owing to preservation method, which ensured that our tissue handling protocols are working properly. There was a significant laboratory effect with respect to cRNA yield, though this effect did not impact sample quality. We conclude that use of consistent protocols for sample collection, RNA extraction, cDNA/cRNA synthesis, labeling, hybridization, platform, image acquisition, normalization, and expression summaries can yield consistent expression values. Moreover, evaluation of sample quality at various steps in the data acquisition process is an important component of a multicenter study to ensure all participating centers adhere to established protocols.

Original languageEnglish (US)
Pages (from-to)34-43
Number of pages10
JournalDiagnostic Molecular Pathology
Volume18
Issue number1
DOIs
StatePublished - Mar 1 2009

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Multicenter Studies
Complementary RNA
RNA
Gene Expression
Analysis of Variance
Complementary DNA

All Science Journal Classification (ASJC) codes

  • Pathology and Forensic Medicine
  • Molecular Biology
  • Cell Biology

Cite this

Archer, K. J., Mas, V., O'Brien, T. R., Pfeiffer, R., Lum, N. L., & Fisher, R. A. (2009). Quality assessment of microarray data in a multicenter study. Diagnostic Molecular Pathology, 18(1), 34-43. https://doi.org/10.1097/PDM.0b013e3181731e14

Quality assessment of microarray data in a multicenter study. / Archer, Kellie J.; Mas, Valeria; O'Brien, Thomas R.; Pfeiffer, Ruth; Lum, Nicole L.; Fisher, Robert A.

In: Diagnostic Molecular Pathology, Vol. 18, No. 1, 01.03.2009, p. 34-43.

Research output: Contribution to journalArticle

Archer, KJ, Mas, V, O'Brien, TR, Pfeiffer, R, Lum, NL & Fisher, RA 2009, 'Quality assessment of microarray data in a multicenter study', Diagnostic Molecular Pathology, vol. 18, no. 1, pp. 34-43. https://doi.org/10.1097/PDM.0b013e3181731e14
Archer, Kellie J. ; Mas, Valeria ; O'Brien, Thomas R. ; Pfeiffer, Ruth ; Lum, Nicole L. ; Fisher, Robert A. / Quality assessment of microarray data in a multicenter study. In: Diagnostic Molecular Pathology. 2009 ; Vol. 18, No. 1. pp. 34-43.
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