Comprehensive detection and serotyping of human adenoviruses by PCR and sequencing

Abstract

Human adenoviruses are common pathogens associated with many diseases, including respiratory, gastrointestinal, and ocular infections. Because they are now being increasingly recognized as agents of life-threatening disseminated infection in immunocompromised patients, robust and sensitive laboratory detection methods are needed for their rapid diagnosis. We describe here a PCR assay using a single primer pair, targeting a region of the hexon gene containing hypervariable region 7, that can detect all known human adenovirus serotypes and allows for serotype determination through the analysis of the nucleotide sequence. This comprehensive assay has proven effective for diagnosing adenoviruses at the serotype level in a broad range of patient specimens, including conjunctival, nasopharyngeal, stool, blood, and urine specimens.

FIG. 1.

A phylogenetic tree from the amino acid sequences of the amplicons of the prototype strains was inferred from an alignment calculated with ClustalX for Windows version 1.81; the tree was built with the program TREECON for Windows version 1.3.b by the neighbor-joining method, as described in the text. Prototype strains are identified by the serotype number at the end of the branches. Serotypes belonging to the same species cluster together; the species are labeled A to F. Bootstrap values (from 1,000 replicates) are expressed as a percentage for each node.

FIG. 2.

A phylogenetic tree from the nucleotide sequences of the amplicons was inferred from an alignment calculated with ClustalX for Windows version 1.81; the tree was built with the program TREECON for Windows version 1.3.b by the neighbor-joining method, as described in the text. Prototype strains are identified by the serotype number at the end of the branches; clinical isolates and strains VR1501 and VR1502 (candidate serotypes 50 and 51) identifiers are highlighted at the end of their branches. Serotypes belonging to the same species cluster together; the species are labeled A to F, with the subspecies B2 also illustrated. Bootstrap values (from 1,000 replicates) are expressed as a percentage for each node.