Detecting and Correcting Batch Effects in High-Throughput Genomic Experiments

Abstract

DETECTING AND CORRECTING BATCH EFFECTS IN HIGH-THROUGHPUT GENOMIC EXPERIMENTS By Sarah Elizabeth Reese A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy at Virginia Commonwealth University Virginia Commonwealth University, 2013 Director: Kellie J. Archer, Ph.D., Associate Professor, Department of Biostatistics; Director, VCU Massey Cancer Center Biostatistics Shared Resource Batch effects are due to probe-specific systematic variation between groups of samples (batches) resulting from experimental features that are not of biological interest. Principal components analysis (PCA) is commonly used as a visual tool to determine whether batch effects exist after applying a global normalization method. However, PCA yields linear combinations of the variables that contribute maximum variance and thus will not necessarily detect batch effects if they are not the largest source of variability in the data. We present an extension of principal components analysis to quantify the existence of batch effects, called guided PCA (gPCA). We describe a test statistic that uses gPCA to test if a batch effect exists. We apply our proposed test statistic derived using gPCA to simulated data and to two copy number variation case studies: the first study consisted of 614 samples from a breast cancer family study using Illumina Human 660 bead-chip arrays whereas the second case study consisted of 703 samples from a family blood pressure study that used Affymetrix SNP Array 6.0. We demonstrate that our statistic has good statistical properties and is able to identify significant batch effects in two copy number variation case studies. We further compare existing batch effect correction methods and apply gPCA to test their effectiveness. We conclude that our novel statistic that utilizes guided principal components analysis to identify whether batch effects exist in high-throughput genomic data is effective. Although our examples pertain to copy number data, gPCA is general and can be used on other data types as well.