Akademik Veri Yönetim Sistemi
REVUE DE MEDECINE VETERINAIRE, cilt.164, sa.7, ss.348-357, 2013 (SCI-Expanded)
The aim of this study was to compare the efficiency of immunohistochemistry, immunofluorescence and in situ PCR to detect viral particles in liver and heart lesions experimentally induced by FadV-4 (Fowl Adenovirus 4) inoculation in broilers. For that, 4 week old healthy broilers were inoculated by FadV-4 (0.5 mL, EID(50)log(2)10(7.3) (Chicken Embryo Infective Dose)/mL) orally (group I, n = 18) or intramuscularly (group II, n = 18) and 12 birds were not inoculated and served as negative controls. Birds were sacrificed 3, 6, 9, 12, 14 and 17 days after; necropsies were systematically performed and liver and heart samples were analysed for histopathology, and for detection of viral antigens using anti FAdV-4 polyclonal chicken antibodies and revelation of the formed immune complexes by rabbit anti chicken IgG and biotinylated goat anti rabbit IgG and horse radish peroxidase (immunohistochemistry) or fluoroisothiocyanate conjugated IgG (immunofluorescence) and using in situ PCR throughout specific amplification of the hexon. Although no mortality was noted, oral and intramuscular inoculations have induced degenerative and inflammatory lesions in heart and liver and the intensity and duration of the organ damage were greater in intramuscularly infected birds. Immunohistochemistry evidenced viral particles in hepatocytes and in some Kupffer cells in liver and in cardiomyocytes in heart as well as in endothelial and mononuclear inflammatory (mainly lymphocytes) cells in all infected birds and the labelling intensity was well correlated to the lesion intensity whereas using immunofluorescence, the antigen distribution was lately evidenced and appeared as attenuated in both organs. The in situ PCR also revealed the virus presence in the same type cells and PCR positivity was highly associated to the lesion intensity and highly corroborated immunohistochemical results. These results show that the in situ PCR method allows the detection of the virus according to the disease extension and suggest that it may be useful to evidence latent infections.