A. Akalu et al., RAPID IDENTIFICATION OF SUBGENERA OF HUMAN ADENOVIRUS BY SEROLOGICAL AND PCR ASSAYS, Journal of virological methods, 71(2), 1998, pp. 187-196
Citations number
31
Categorie Soggetti
Virology,"Biochemical Research Methods","Biothechnology & Applied Migrobiology
Bacterially expressed recombinant protein IX (pIX) of human adenovirus
serotype 2 (Ad2) and 3 (Ad3) was evaluated for use as a subgenus-spec
ific antigen by enzyme-linked immunosorbent assay (ELISA) and Western
blotting. Patients sera positive by ELISA for the genus-specific adeno
virus hexon antigen recognized the recombinant pIX of Ad2 and Ad3 in a
subgenus-specific manner by both assays. Polyclonal rabbit serum rais
ed against the recombinant Ad2pIX reacted strongly by indirect immunof
luorescence assay, with Adl, 2 and 5 (subgenus C) but not with serotyp
es representing other subgenera. In a similar way, anti-Ad3pIX reacted
with Ad3, 7, 11 and 14 (subgenus B), but not with serotypes represent
ing other subgenera. A polymerase chain reaction showed that the compl
ete pIX gene could be amplified in a subgenus specific fashion using p
rimers specific for Ad3 (subgenus B), Ad2 (subgenus C), or Ad40/41 (su
bgenus F). The pIX gene from the available isolates of subgenus A, D a
nd E was not amplified with these primers. The use of pIX-based serolo
gical assays is useful For subgenotyping as a primary screen of anti-A
d sera. It is much more rapid than the currently used neutralization a
ssay 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. (C) 1998 E
lsevier Science B.V. All rights reserved.