Molecular typing of human adenoviruses by PCR and sequencing of a partial region of the hexon gene

Abstract

Human adenoviruses (Ads) are responsible for a substantial disease burden. Type-specific identification of Ads can help guide therapeutic and disease prevention strategies and aid epidemiological investigations. Immunotyping of Ads by serum neutralization (SN) is laborious and time consuming and depends upon type-specific antisera that are in short supply. A rapid molecular typing assay based on polymerase chain reaction (PCR) amplification and sequencing of Ad hexon gene hyper-variable regions 1–6 (HVR_1–6) known to contain type-specific epitopes was evaluated as an alternative to SN. Deduced amino acid sequences of HVR_1–6 obtained from all 51 currently recognized Ad prototype strains were well resolved, with the exception of types 15 and 29, which were identical. Of 192 temporally and geographically diverse Ad field isolates sequenced in this study, and 111 previously published sequences, all more closely matched their predicted prototype strains. Ads were also detected and correctly identified directly from 24 clinical specimens positive by culture or antigen detection. PCR and sequencing of HVR_1–6 offers a practical alternative to SN for typing most Ads and can be readily adapted for use in laboratories with molecular capabilities.