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Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs
Published on: April 14, 2015
Assessing Agreement between miRNA Microarray Platforms
Niccolò P Bassani1, Federico Ambrogi2, Elia M Biganzoli1
1Department of Clinical Sciences and Community Health, University of Milan, Via Vanzetti 5 20133, Milano (MI), Italy.
Abstract:
Over the last few years, miRNA microarray platforms have provided great insights into the biological mechanisms underlying the onset and development of several diseases. However, only a few studies have evaluated the concordance between different microarray platforms using methods that took into account measurement error in the data. In this work, we propose the use of a modified version of the Bland-Altman plot to assess agreement between microarray platforms. To this aim, two samples, one renal tumor cell line and a pool of 20 different human normal tissues, were profiled using three different miRNA platforms (Affymetrix, Agilent, Illumina) on triplicate arrays. Intra-platform reliability was assessed by calculating pair-wise concordance correlation coefficients (CCC) between technical replicates and overall concordance correlation coefficient (OCCC) with bootstrap percentile confidence intervals, which revealed moderate-to-good repeatability of all platforms for both samples. Modified Bland-Altman analysis revealed good patterns of concordance for Agilent and Illumina, whereas Affymetrix showed poor-to-moderate agreement for both samples considered. The proposed method is useful to assess agreement between array platforms by modifying the original Bland-Altman plot to let it account for measurement error and bias correction and can be used to assess patterns of concordance between other kinds of arrays other than miRNA microarrays.
Insights
This study introduces a modified Bland-Altman plot to evaluate agreement between miRNA microarray platforms. The method revealed good concordance for Agilent and Illumina, but poor-to-moderate agreement for Affymetrix.
Area of Science:
- Biotechnology
- Genomics
- Medical Diagnostics
Background:
- MicroRNA (miRNA) microarrays are crucial for understanding disease mechanisms.
- Limited studies assess concordance between miRNA microarray platforms considering measurement error.
Purpose of the Study:
- To propose and validate a modified Bland-Altman plot for assessing agreement between miRNA microarray platforms.
- To evaluate the concordance of Affymetrix, Agilent, and Illumina miRNA platforms.
Main Methods:
- Profiling of a renal tumor cell line and normal human tissues on three miRNA platforms (Affymetrix, Agilent, Illumina) in triplicate.
- Assessment of intra-platform reliability using concordance correlation coefficients (CCC and OCCC).
- Application of a modified Bland-Altman plot to analyze inter-platform agreement, accounting for measurement error and bias.
Main Results:
- All platforms demonstrated moderate-to-good intra-platform repeatability (CCC and OCCC).
- Modified Bland-Altman analysis showed good concordance for Agilent and Illumina platforms.
- Affymetrix platform exhibited poor-to-moderate agreement between samples.
Conclusions:
- The modified Bland-Altman plot effectively assesses agreement between miRNA microarray platforms, accounting for measurement error.
- Agilent and Illumina platforms show better concordance compared to Affymetrix for miRNA profiling.
- The method is adaptable for evaluating concordance across various array-based technologies.

