Type‐specific identification of human adenovirus 3, 7, and 21 by a multiplex PCR assay†

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.

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

“…Besides the 15 viral isolates, other eight nasopharyngeal samples negative for viral isolation in cell culture were positive for AdV by PCR detection. This finding agrees with previous studies which have shown that even though viral culture has been considered as the "gold standard" for AdV diagnosis, the PCR-based assays are more sensitive for this purpose (Allard et al 1990, Pring-Åkerblom & Adrian 1994, Alistair et al 1996, Kajiwara et al 1999, Xu et al 2000, Avellón et al 2001, Echavarria et al 2001, Heim et al 2003.…”

“…Immunofluorescence and other immunodiagnostic methods using direct antigen detection are faster than culture, but they lack sensitivity (Minnich & Ray 1980, Mahafzah & Landry 1989, Portillo & Cruz 2000. Recently polymerase chain reaction (PCR)-based assays have proven to be useful for AdV detection, showing higher specificity and sensitivity in comparison to the classic methods (Allard et al 1990, Pring-Åkerblom & Adrian 1994, Alistair et al 1996, Kajiwara et al 1999, Xu et al 2000, Avellón et al 2001, Echavarria et al 2001, Heim et al 2003.…”

Hospitalization rates for human metapneumovirus infection among 0- to 3-year-olds in Gipuzkoa (Basque Country), Spain

“…RSV, influenza A and B, parainfluenza viruses types 1-3 and adenovirus were investigated through culture using the rapid shell vial techniques on MDCK, A-549 and LLC-MK2 cell lines. Similarly, the presence of RSV [11], influenza A and B [11,12], parainfluenza 1-4 [11], rhinovirus [13], respiratory coronaviruses [14], and coronavirus NL63 [15] were investigated using multiplex reverse transcriptase-polymerase chain reaction (RT-PCR) methods, the presence of adenovirus was investigated with a PCR method [16]. RT-PCR for HMPV was performed using primers derived from the F gene [17].…”

Monitoring of adenovirus load in stool by real-time PCR permits early detection of impending invasive infection in patients after allogeneic stem cell transplantation

“…They have been divided into six major subgroups (subgenera or species A-F) on the basis of the oncogenic, hemagglutinating, morphological and DNA sequence properties. [8][9][10][11][12] Infections can cause localized disease such as enteritis, upper respiratory tract infection, encephalitis or cystitis. [13][14][15] However, AdV infections in immunocompromised patients tend to become invasive, and disseminated disease is associated with very high mortality.…”