Complete genome sequencing of a chicken embryo egg-adapted Duck Atadenavirus A

The virus was propagated into allantoic sac of 10-day-old specific-pathogen-free embryonated chicken eggs. The embryos were incubated at 37 C for seven days and examined daily for their viability. After harvesting the allantoic fluid aseptically from eggs, seven blind passages were performed through allantoic cavity into another fresh set of embryonated eggs until the high haemagglutination (HA) titer was observed. Then, after extraction of viral DNA from the allantoic fluid, overlapping fragments of the viral genome sequence were generated by PCR using 25 pairs of primers. Purified PCR products were subjected to complete genome sequencing by next generation sequencing approach.