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Updated: Jan 2, 2026
Author Spotlight: Advancements in Multiplex Detection of Respiratory Viruses
Published on: November 10, 2023
Type-specific identification of human adenovirus 3, 7, and 21 by a multiplex PCR assay
1Centers for Disease Control and Prevention, Division of Viral and Rickettsial Diseases, Respiratory and Enteric Viruses Branch, Atlanta, Georgia, USA.
Abstract:
Human adenovirus (Ad) serotypes 3, 7, and 21 of DNA cluster B:1 are often associated with severe respiratory illness, particularly in infants and young children and, in addition to Ad4, are among the most important causes of acute respiratory disease syndrome in new military recruits. To address the inherent problems associated with classic typing methods, we developed a multiplex PCR assay for the rapid, specific identification of Ad3, Ad7, and Ad21 field isolates. To design type-specific primers for our assay, we sequenced the Ad21 hexon gene and compared this sequence with previously published sequences of Ad3, Ad7, and Ad16. The overall nucleotide (nt) and amino acid (aa) identities between Ad21 and Ad3, Ad7, and Ad16 were similar (ranges 78.3-80.8% nt; 84.1-86.2% aa), with significantly greater variability in the regions of the hexon that encode surface loops 1 and 2. Type-specific primers designed to the hypervariable regions correctly identified Ad3, Ad7, and Ad21 prototype strains and 53 previously typed Ad field isolates. No cross-reactions with other Ad serotypes were identified. Our multiplex PCR assay for type-specific identification of Ad3, Ad7, and Ad21 isolates will provide a rapid and convenient tool for the epidemiologic investigation of Ad-associated respiratory illness.
Insights
A new multiplex PCR assay rapidly identifies human adenovirus (Ad) serotypes 3, 7, and 21, common causes of severe respiratory illness. This tool aids in the epidemiologic investigation of adenovirus-associated respiratory disease.
Area of Science:
- Virology
- Molecular Biology
- Epidemiology
Background:
- Human adenovirus (Ad) serotypes 3, 7, and 21 are linked to severe respiratory illnesses in children and military recruits.
- Traditional adenovirus typing methods present inherent challenges for rapid identification.
Purpose of the Study:
- To develop a rapid and specific multiplex PCR assay for identifying Ad3, Ad7, and Ad21 field isolates.
- To improve the epidemiologic investigation of adenovirus-associated respiratory illness.
Main Methods:
- Sequencing of the Ad21 hexon gene and comparison with Ad3, Ad7, and Ad16 sequences.
- Design of type-specific primers targeting hypervariable regions of the hexon gene.
- Validation of the multiplex PCR assay using prototype strains and 53 field isolates.
Main Results:
- Nucleotide and amino acid identities between Ad21 and related serotypes ranged from 78.3-80.8% nt and 84.1-86.2% aa.
- Hypervariable regions in the hexon gene showed significant variability, enabling type-specific primer design.
- The assay correctly identified Ad3, Ad7, and Ad21 strains with no cross-reactions with other serotypes.
Conclusions:
- The developed multiplex PCR assay offers a rapid and convenient method for the type-specific identification of Ad3, Ad7, and Ad21.
- This assay will be a valuable tool for epidemiological studies of adenovirus-related respiratory diseases.
