Systematic evaluation of three microRNA profiling platforms: microarray, beads array, and quantitative real-time PCR array

PLoS One. 2011 Feb 11;6(2):e17167. doi: 10.1371/journal.pone.0017167.

Abstract

Background: A number of gene-profiling methodologies have been applied to microRNA research. The diversity of the platforms and analytical methods makes the comparison and integration of cross-platform microRNA profiling data challenging. In this study, we systematically analyze three representative microRNA profiling platforms: Locked Nucleic Acid (LNA) microarray, beads array, and TaqMan quantitative real-time PCR Low Density Array (TLDA).

Methodology/principal findings: The microRNA profiles of 40 human osteosarcoma xenograft samples were generated by LNA array, beads array, and TLDA. Results show that each of the three platforms perform similarly regarding intra-platform reproducibility or reproducibility of data within one platform while LNA array and TLDA had the best inter-platform reproducibility or reproducibility of data across platforms. The endogenous controls/probes contained in each platform have been observed for their stability under different treatments/environments; those included in TLDA have the best performance with minimal coefficients of variation. Importantly, we identify that the proper selection of normalization methods is critical for improving the inter-platform reproducibility, which is evidenced by the application of two non-linear normalization methods (loess and quantile) that substantially elevated the sensitivity and specificity of the statistical data assessment.

Conclusions: Each platform is relatively stable in terms of its own microRNA profiling intra-reproducibility; however, the inter-platform reproducibility among different platforms is low. More microRNA specific normalization methods are in demand for cross-platform microRNA microarray data integration and comparison, which will improve the reproducibility and consistency between platforms.

Publication types

  • Evaluation Study

MeSH terms

  • Cell Transformation, Neoplastic
  • Gene Expression Profiling / methods*
  • Humans
  • MicroRNAs / genetics*
  • Microspheres*
  • Oligonucleotide Array Sequence Analysis / methods*
  • Oligonucleotides / genetics
  • Osteosarcoma / genetics
  • Osteosarcoma / pathology
  • Real-Time Polymerase Chain Reaction / methods*
  • Reproducibility of Results

Substances

  • MicroRNAs
  • Oligonucleotides
  • locked nucleic acid