tailieunhanh - Báo cáo khoa học: "Optimization of in situ hybridization assay using nonradioactive DNA probes for the detection of canine erpesvirus (CHV) in paraffin-embedded sections"

Tuyển tập các báo cáo nghiên cứu khoa học quốc tế về bệnh thú y đề tài: Optimization of in situ hybridization assay using nonradioactive DNA probes for the detection of canine erpesvirus (CHV) in paraffin-embedded sections | J. Vet. Sci. 2004 5 1 71-73 JOURNAL OF Short Communication Veterinary Science Optimization of in situ hybridization assay using non-radioactive DNA probes for the detection of canine herpesvirus CHV in paraffin-embedded sections Okjin Kim Department of Laboratory Animal Sciences and Center for Animal Resource Development College of Medicine Seoul National University Seoul 110-799 Korea Two non-radioactive probes using digoxigenin or biotin were developed for detecting canine herpesvirus CHV and compared for their sensitivities by in situ hybridization ISH in formalin fixed paraffin embedded sections which has been used routinely in veterinary fields. Sections of the CHV-infected cell preparation were subjected to several different ISH protocols using digoxigenin- or biotin-labeled probe respectively. Results were compared for the hybridization and background signal intensities. The best result was obtained by the optimized ISH protocol using digoxigenin-labeled probe for detection of CHV DNA. The optimized ISH assay which developed in this study may be a valid tool for the study of pathogenesis and diagnosis of CHV infection. Key words canine herpesvirus digoxigenin biotin in situ hybridization Canine herpesvirus CHV is a member of the alpha herpesvirus subfamily that can cause a severe hemorrhagic disease in neonatal pups as well as mild or subclinical respiratory infections in adult dogs 1 . Since its isolation CHV has been identified in many countries and a worldwide distribution is presumed. Several studies in South Korea 5 9 and European countries 6 7 suggest a high prevalence of the virus among the dog population. As CHV is presumed to be widespread among the dog population and as the economic losses that breeding kennels may suffer after infection with CHV may be disastrous it seems necessary to determine the CHV infection more exactly. Formalin-fixed paraffin-embedded tissues have been used routinely in veterinary practice. For the study of pathogenesis and