This technology includes a real-time polymerase chain reaction (PCR) assay using hydrolysis probe (TaqMan®) chemistry which was developed for detection and type-specific identification of human adenovirus type 4 (HAd4) genomic DNA in clinical specimens. The assay uses unique oligonucleotide primer and probes designed to conserved regions of the HAd4 hexon protein gene. Assay reagents and amplification conditions were optimized and sensitivity and specificity characteristics determined with DNA extracts from multiple HAd4 laboratory strains and other respiratory viral pathogens.