Species-specific identification of human adenoviruses by a multiplex PCR assay

A multiplex PCR assay was developed by using primers to the fiber gene that could differentiate human adenovirus (Ad) species A through F in a single amplification reaction, The assay correctly identified the species of all 4 9 recognized Ad prototype strains as well as 180 geographically and tempora lly diverse Ad field isolates. Ad serotype 6 (Ad6) (species C), Ad16 (speci es B), Ad31 (species A), and Ad40 and Ad41 (species F) could also be distin guished by amplicon size within each respective species. In comparison, a p reviously described Ad species-specific multiplex PCR assay that used prime rs to the Ad hexon gene gave equivocal results with several serotypes of sp ecies B, whereas our multiplex assay amplified all species B serotypes equa lly well. Our multiplex PCR assay will permit rapid, accurate, and cost-eff ective classification of Ad isolates.