Rapid identification of subgenera of human adenovirus by serological and PCR assays

Bacterially expressed recombinant protein IX (pIX) of human adenovirus serotype 2 (Ad2) and 3 (Ad3) was evaluated for use as a subgenus-specific antigen by enzyme-linked immunosorbent assay (ELISA) and Western blotting. Patients sera positive by ELISA for the genus-specific adenovirus hexon antigen recognized the recombinant pIX of Ad2 and Ad3 in a subgenus-specific manner by both assays. Polyclonal rabbit serum raised against the recombinant Ad2pIX reacted strongly by indirect immunofluorescence assay, with Adl, 2 and 5 (subgenus C) but not with serotypes representing other subgenera. In a similar way, anti-Ad3pIX reacted with Ad3, 7, 11 and 14 (subgenus B), but not with serotypes representing other subgenera. A polymerase chain reaction showed that the complete pIX gene could be amplified in a subgenus specific fashion using primers specific for Ad3 (subgenus B), Ad2 (subgenus C), or Ad40/41 (subgenus F). The pIX gene from the available isolates of subgenus A, D and E was not amplified with these primers. The use of pIX-based serological assays is useful for subgenotyping as a primary screen of anti-Ad sera. It is much more rapid than the currently used neutralization assay or hemagglutination inhibition test. The application of anti-pIX sera by immunofluorescence and a pIX gene-based PCR are rapid methods which will improve subgenus identification of adenoviruses.