tailieunhanh - Báo cáo khoa học: "Detection of canine distemper virus (CDV) through one step RT-PCR combined with nested PCR"

Tuyển tập các báo cáo nghiên cứu khoa học quốc tế về bệnh thú y đề tài: Detection of canine distemper virus (CDV) through one step RT-PCR combined with nested PCR | J. Vet. Sci. 2001 2 1 59-63 JOURNAL OF Veterinary Science Detection of canine distemper virus CDV through one step RT-PCR combined with nested PCR Yong-Hwan Kim Kyu-Woan Cho1 Hwa-Young Youn Han Sang Yoo and Hong-Ryul Han Department of Internal Medicine and Infectious Diseases College of Veterinary Medicine and School of Agricultural Biotechnology Seoul National University Seoul 151-742 Korea 1 Department of Internal Medicine College of Veterinary Medicine Chungnam National University Taejon 305-764 Korea A one step reverse transcription PCR RT-PCR combined nested PCR was set up to increase efficiency in the diagnosis of canine distemper virus CDV infection after developement of nested PCR. Two PCR primer sets were designed based on the sequence of nucleocapsid gene of CDV Onderstepoort strain. One-step RT-PCR with the outer primer pair was revealed to detect 102 PFU m . The sensitivity was increased hundredfold using the one-step RT-PCR combined with the nested PCR. Specificity of the PCR was also confirmed using other related canine virus and peripheral blood mononuclear cells PBMC and body secretes of healthy dogs. Of the 51 blood samples from dogs clinically suspected of CD 45 samples were revealed as positive by one-step RT-PCR combined with nested PCR. However only 15 samples were identified as positive with a single one step RT-PCR. Therefore approximately 60 increase in the efficiency of the diagnosis was observed by the combined method. These results suggested that one step RT-PCR combined with nested PCR could be a sensitive specific and practical method for diagnosis of CDV infection. Key words CDV RT-PCR nested PCR Introduction Canine distemper CD is a worldwide highly contagious disease in young dogs particularly in 3 to 6 months of age with high morbidity and mortality. It is manifested by a diphasic fever curve and acute rhinitis and later by bronchitis catarrhal pneumonia severe gastroenteritis and nervous signs 17 . The disease spread mainly in the