鸭黄病毒SDbz株的分离与初步鉴定

鸭黄病毒病为近年来新发的鸭病,给养鸭业带来了极大的经济损失。为了深入研究本病的防制,本试验对鸭黄病毒(duck flavivirus,DFV)进行了分离鉴定。取疑似感染DFV的病鸭病料,经细菌分离初步排除细菌感染后,应用RT-PCR检测呈现DFV阳性,处理后将其接种到鸭胚成纤维细胞(DEF)和健康鸭胚上进行病毒分离传代。结果显示,在DEF细胞上第1代48 h就开始出现CPE,随着时间的延长CPE更加明显,通常在72~96 h产生典型CPE;接种鸭胚每一代均出现死亡,且死亡时间多集中于接种后60~72 h,死亡鸭胚胚体水肿、出血、发育不良、胚肝严重出血、肿胀或斑驳样坏死等病变。将病毒DEF细胞和鸭胚分离物应用血凝试验、毒价测定、病毒中和试验、RT-PCR及人工感染试验进行检测鉴定,证明所分离到的病毒为DFV,并将其命名为DFV SDbz株。

Isolation and Identification of Duck Flavivirus SDbz Strain

Duck flavivirus(DFV)disease was a kind of new duck disease. It had brought great economic losses to the duck industry in recent years. In order to study thoroughly and prevent the disease,it was isolated and identified as follows.The bacterial infection was initially ruled out from the samples by bacteria isolation. The samples showed DFV positive by RT-PCR detection. Then they were inoculated into DEF cells and healthy duck embryo for virus isolation and passages. Cytopathic effect (CPE) in DEF cells were observed in the first generation in the 48 h after inoculation, CPE was more evident with the extension of the time, and the typical CPE were observed as from 72 to 96 h after inoculation. Each generation of inoculated duck embryo appeared to be death, and the time of death were more concentrated in 60 to 72 h after inoculation. The dead duck embryo body were observed as edema, hemorrhage, dysphasia, severe hepatic hemorrhage, swelling or necrosis mottled. The virus isolated in DEF cells and duck embryo were detected and identified by hemagglutination test, titer determination, virus neutralization test, RT-PCR and artificial infection test. The results showed that the virus isolated was DFV and named as DFV SDbz strain.