FIG. 3.

Detection of Ad41 mRNA by RT-PCR assay and subsequent nested PCR assay with hexon primer sets at different MOIs of Ad41 and different times p.i. (A) Lanes: 1 to 4, RT-PCR assay of mRNA from G293 cells infected with 5 × 10⁴ IU of Ad41 at 0, 1, 3, and 5 days p.i.; 5 to 8, RT-PCR assay of mRNA from G293 cells infected with 5 × 10² IU of Ad41 at 0, 1, 3, and 5 days p.i.; 9 to 12, RT-PCR assay of mRNA from G293 cells infected with 50 IU at 0, 1, 3, and 5 days p.i.; 13 to 16, RT-PCR assay of mRNA from G293 cells infected with 5 IU at 0, 1, 3, and 5 days p.i.; 17, negative control; 18, positive control. (B) Lanes: 1 to 4, nested PCR assay of mRNA from G293 cells infected with 5 × 10⁴ IU of Ad41 at 0, 1, 3, and 5 days p.i.; 5 to 8, nested PCR assay of mRNA from G293 cells infected with 5 × 10² IU of Ad41 at 0, 1, 3, and 5 days p.i.; 9 to 12, nested PCR assay of mRNA from G293 cells infected with 50 IU at 0, 1, 3, and 5 days p.i.; 13 to 16, nested PCR assay of mRNA from G293 cells infected with 5 IU at 0, 1, 3, and 5 days p.i.; 17, negative control; 18, positive control. A 100-bp DNA ladder was used as a molecular size marker (lane M). The detection sign (+ or −) was based on visual examination of the agarose gel for an amplicon of the correct size.