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 lifethreatening 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.

Antigenic and genomic characterization of adenovirus associated to respiratory infections in children living in Northeast Brazil

“…Polymerase chain reaction (PCR), real-time PCR and immunochromatographic kits have recently become available for this purpose (Fujimoto et al 2004). Typing of Ads isolates can be achieved by neutralization, hemmaglutination inhibition, restriction endonuclease analysis (REA), sequencing and, PCR combined with REA (Allard et al 2001, Sarantis et al 2004. Until recently, the interest in serotype and genotype determination of Ad isolates was mainly epidemiological, however, this has changed due to the increasingly clear association between specific serotypes and genotypes and more severe disease presentation (Kajon & Wadell 1994, Larrañaga et al 2000, Carballal et al 2002.…”

Adenoviruses C in non-hospitalized Mexican children older than five years of age with acute respiratory infection

“…To circumvent practical problems associated with traditional serum neutralization studies molecular typing methods have been proposed (Kidd et al1996, Li et al 1999, Elnifro et al 2000, such as, restriction endonuclease analysis of full-length adenovirus DNA or by PCR using typespecific primers (Garnet et al 2002). Other approaches have been the PCR amplification of the hypervariable region of the hexon gene followed by DNA sequencing (Takeuchi et al 1999, Sarantis et al 2004, PCR amplification followed by restriction endonuclease analysis, and multiplex PCR-enzyme hybridization methods with species-specific-oligonucleotide primers and probes (Gu et al 2003, Pehler-Harrington et al 2004, Echavarria et al 2006). …”

Molecular characterization of adenoviruses from children presenting with acute respiratory disease in Uberlândia, Minas Gerais, Brazil, and detection of an isolate genetically related to feline adenovirus

“…In general, neutralisation or heamagglutination inhibition assays are used for HAdV serotyping (Swenson et al 2003). However, serotyping can be done using immunofluorescence assay (IF) with serotype-specific monoclonal antibodies (Wood et al 1997), genus-specific polymerase chain reaction (PCR) combined with restriction endonuclease digestion (PCR-restriction fragment length polymorphism) analysis (Allard et al 2001) and sequence analysis (Sarantis et al 2004).…”