The design and analysis of microarray experiments: applications in parasitology.
Microarray experiments can generate enormous amounts of data, but large datasets are usually inherently complex, and the relevant information they contain can be difficult to extract. For the practicing biologist, we provide an overview of what we believe to be the most important issues that need to be addressed when dealing with microarray data. In a microarray experiment we are simply trying to identify which genes are the most "interesting" in terms of our experimental question, and these will usually be those that are either overexpressed or underexpressed (upregulated or downregulated) under the experimental conditions. Analysis of the data to find these genes involves first preprocessing of the raw data for quality control, including filtering of the data (e.g., detection of outlying values) followed by standardization of the data (i.e., making the data uniformly comparable throughout the dataset). This is followed by the formal quantitative analysis of the data, which will involve either statistical hypothesis testing or multivariate pattern recognition. Statistical hypothesis testing is the usual approach to "class comparison," where several experimental groups are being directly compared. The best approach to this problem is to use analysis of variance, although issues related to multiple hypothesis testing and probability estimation still need to be evaluated. Pattern recognition can involve "class prediction," for which a range of supervised multivariate techniques are available, or "class discovery," for which an even broader range of unsupervised multivariate techniques have been developed. Each technique has its own limitations, which need to be kept in mind when making a choice from among them. To put these ideas in context, we provide a detailed examination of two specific examples of the analysis of microarray data, both from parasitology, covering many of the most important points raised.
Morrison
David A
DA
Department of Parasitology (SWEPAR), National Veterinary Institute and Swedish University of Agricultural Sciences, Uppsala, Sweden.
Ellis
John T
JT
D000704:Analysis of Variance; D000818:Animals; D020224:Expressed Sequence Tags; D007090:Image Interpretation, Computer-Assisted; D015999:Multivariate Analysis; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D010274:Parasitology / Q000379:methods*; D012107:Research Design; D014122:Toxoplasma / Q000235:genetics / Q000502:physiology