Quantification of viral genomes by qPCR to confirm OAd replication in PDOs
PDOs were infected with 2 different OAds (OAd1 and OAd2) at 2 MOI, subjected to freeze-thaw cycles and to DNA extraction (passage 1, P1). Fifteen μL of lysate-derived supernatants were used to infect fresh PDOs, and the same process was repeated to quantify viral genomes in organoids (passage 2, P2). Adenoviral genomes were quantified by qPCR amplification of hexon genes using a standard curve consisting of DNA dilutions of known copy numbers (102–107). High viral yields were detected in P1 and P2 with both viruses, proving that the OAd effectively replicates within PDOs. Data are represented as mean ± SEM (n = 3).