Detection of Fowl Adenovirus DNA from Formalin-Fixed and Paraffin-Embedded Sections by PCR and Classification of Serotypes by Sequencing of PCR Products

Abstract

Detection of fowl adenovirus (FAV) DNA from formalin-fixed and paraffin-embedded (FFPE) sections was attempted by PCR. Serotypes of FAV were classified by sequencing the PCR products. In trials of PCR using a positive control infected with serotype 2 FAV, the best primer set was 57F forward primer (5'-CAARTTCAGRCAGACGGT-3') and 26R reverse primer (5'-GGCTTGACGTACGCTCCGTA-3'). A second PCR with the same primer set revealed a clearer band in the electrophoresis of generated PCR products. Generated PCR products were confirmed to be derived from infected FAV. In addition, PCR and sequencing of PCR products of the liver FFPE sections, from two natural inclusion body hepatitis cases that were not examined for virologic isolation, suggested that the detected FAV was serotype 8a. The PCR of FFPE sections, and serotyping by the sequencing of PCR products, are useful for diagnosis and epidemiologic analysis of FAV infections.

Propagation and Molecular Characterization of Fowl Adenovirus Serotype 8b Isolates in Chicken Embryo Liver Cells Adapted on Cytodex™ 1 Microcarrier Using Stirred Tank Bioreactor

“…The amplification of the hexon gene of UPM08136CELP5B1 and UPM08136CELP20B1 yielded a partial hexon DNA 738-750 bp in size, which corresponds to the report of Meulemans et al [29] who initially published HexA1/HexB1 primers. The primer had been used by several researchers for identification and studies of FAdV [43][44][45][46][47][48]. The amplification of the fiber gene using the novel fbrF/fbrR primers yielded partial fiber gene, mainly the knob region.…”