Pharyngitis caused by group A streptococci (GAS) is one of the most common infections around the world. However, relatively little is known about which genes are expressed and which genes regulate expression during acute infection. Due to their ability to provide genome-wide views of gene expression at one time, microarrays are increasingly being incorporated in GAS research. In this study, a novel electrochemical detection-based microarray was used to identify gene expression patterns among humans with culture-confirmed GAS pharyngitis. Using 14 samples (11 GAS-positive and three GAS-negative) obtained from subjects seen at the Brooke Army Medical Center paediatric clinic, this study demonstrated two different clusters of gene expression patterns. One cluster expressed a larger number of genes related to phages, immune-system evasion and survival among competing oral flora, signifying a potentially more virulent pattern of gene expression. The other cluster showed a greater number of genes related to nutrient acquisition and protein expression. This in vivo genome-wide analysis of GAS gene expression in humans with pharyngitis evaluated global gene expression in terms of virulence factors.