A rooted phylogenetic analysis of the viral family Picornaviridae identified several major clusters within this viral family. The genus Parechovirus, including Ljungan virus, was found to be basal to the family. Within the family, there were two major subgroups: (1) a cluster including viruses assigned to the genera Enterovirus and Rhinovirus; and (2) a cluster including Teschovirus, Cardiovirus, Erbovirus, Aphthovirus, and probably Kobuvirus. Within the former cluster, the genera Enterovirus and Rhinovirus were found not to be monophyletic. Principal components analysis was used to identify different patterns with regard to the relative sequence divergence of the 11 polyproteins encoded by the picornavirus genome. In comparisons of closely related pairs of viruses, the nonstructural proteins were generally more conserved than the capsid proteins 1B, 1C, and 1D. This pattern is most likely explained by the predominance of host antibody-driven selection on the differentiation of closely related viral genomes. However, the same pattern was not seen in more distantly related pairs of viruses, which showed a number of distinct patterns with regard to the relative conservation of the 11 proteins.