Evaluation of a recombinant adenovirus as an internal process control virus

BLANCO FERNANDEZ, MARIA DOLORES; MELINA ELIZABETH BARRIOS; ROBERTINA VIVIANA CAMMARATTA; CAROLINA TORRES; VIVIANA A. MBAYED

Salta, Salta

Simposio; III Simposio Latinoamericano de Virologia Ambiental; 2016

Universidad Nacional de Salta

A recombinant non-replicative adenovirus (rAdV-IPC), carrying agenetic construction, was evaluated as an internal process control(IPC) for virus concentration procedures from tap water andsewage. rAdV-IPC was obtained and produced by transfectionand successive passages in 293A cells. Infectivity of rAdV-IPC,huAdV3 and bacteriophage PP7 at different pH values (3, 7 and10) was evaluated and compared. No significant differences ininfectious virus titers were observed for both adenoviruses afterincubation at each pH condition. In contrast, a significantdecrease was observed for PP7 under treatment at pH 3 and 10.The performance of rAdV-IPC and PP7 during virus concentrationby ultrafiltration and ultracentrifugation was evaluated andcompared. Viruses were quantified along the concentrationprocess using qPCR and in vitro culture assays, to calculate totalvirus and infectious virus recovery. Recovery was significantlylower when comparing infectious versus total virus for both controlviruses. While no significant differences in recovery between IPCviruses were observed for the ultracentrifugation process,differential behavior was observed for ultrafiltration recovery. Theability of IPC virus to mimic the behavior of the target virus shouldbe analyzed for each particular virus and detection procedure.