Development of a PCR-based dot blot assay for the detection of fowl adenovirus

Abstract

Group-I Fowl adenovirus ( FAdV ) is still widespread in China's chicken farms, leading to huge economic losses. The traditional PCR method, which can detect all serotypes at the same time, is not sensitive enough to obtain accurate results, especially in some samples containing only a low titer of virus, such as contaminated live vaccine. In order to solve this problem, this study developed a dot blot assay based on the above PCR method. A total of 6 probes targeting the conserved region of FAdV were designed and systematically optimized through sensitivity, accuracy, and stability analyses. Results showed that it is not only suitable for 12 serotypes, but also effectively improve the sensitivity, which increased more than 100 times in comparison with PCR assay. Moreover, this sensitivity was increased 100 times when detecting contaminated live vaccine samples, showing the great prospect of this method in daily monitoring.

FAdV-4 Promotes Expression of Multiple Cytokines and Inhibits the Proliferation of aHEV in LMH Cells

“…202110889250.5). Primers and probes for fluorescent RAA and RAA-LFD detection were designed and synthesized from the conserved domains of 12 serotype reference strains of FAdV-I [ 17 ]. Sensitivity tests for the standard plasmid demonstrated that the minimum detectable concentration of fluorescent RAA and RAA-LFD was 100 copies/µL.…”