Purified HAdV5-PUMA in PBS were dissociated by incubation with 0.05% deoxycholate at 56 °C for 2 mins, while BVCAR virions in PBS were dissociated by a treatment of four cycles of freeze-thawing. Microtitre plates were coated with 100 μl of the viral proteins (1 μg ml−1) diluted in PBS and left overnight at 4 °C in a moisture chamber. The coated wells were washed four times with PBS and incubated with 200 μl blocking solution (2% bovine serum albumin in TBS) for 1 h at RT. After washing, 100 μl of each sera was added to the wells and incubated for 1 h at RT. Plates were then washed and incubated with anti-rat-horseradish peroxidase diluted to 1: 15,000 in blocking solution for 1 h at RT. After washing, 100 μl of OPD (o-phenylenediamine, Sigma-Aldrich) substrate was added, and the reaction was blocked by addition of 1 N HCl. OD was measured at 450 nm using an ELISA plate reader (Wallac 1420 Multilabel Counter, PerkinElmer).