The reaction consisted of 0.1 M DDT (Invitrogen), 1X Platinum Quantitative PCR Supermix-UDG (Invitrogen), Pan-HBoV-F and Pan-HBoV-R primers each at a concentration of 100 mm, Pan-HBoV-NS1 probe at 10 mm concentration, ROX 25 mm (Invitrogen) 2.5 ml of template DNA and RNase free water to a final reaction volume of 25 ml. The amplification consisted of an initial denaturation at 95uC for 10 min, followed by 40 cycles with denaturation at 95uC for 15 sec, annealing at 55uC for 30 sec and extension at 60uC for 45 sec.