鹿粘膜病病毒的分离鉴定  

Isolation and identification of deer mucosal disease virus

作　　者：杜锐[1] 王全凯[1] 王树志[1] 王新平[1] 任东波[1] 

机构地区：[1]吉林农业大学中药材学院,吉林长春130118

出　　处：《中国预防兽医学报》2004年第5期376-379,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(30300256);吉林省科技发展计划项目(20030552-2)

摘　　要：本研究首次应用MDBK传代细胞从腹泻幼鹿粪样中分离获得一株病毒(MDV N71)。该毒株可使培养细胞出现明显的空泡变性,电镜负染观察接毒细胞培养物见有典型的粘膜病病毒粒子。MDV N71在MDBK传代细胞上的TCID_(50)为10^(-5.4),对乙醚、氯仿和胰蛋白酶敏感,56℃70min被灭活。对酸的耐受力较弱,在pH3.0环境下毒力明显降低。对碱有较强的耐受力,在pH9.0环境下依然稳定。不凝集绵羊、猪、兔和鸡红细胞。用MDV长春184毒株的特异性免疫荧光抗体染色(IFA),可以在感染细胞浆内见有特异性荧光。该毒株的致细胞病变作用,可被MDV长春184毒株抗血清所中和。提取病毒感染细胞的总RNA进行反转录聚合酶链反应(RT-PCR),结果扩增出的片段大小为400 bp。根据MDV N71的理化特性、形态学、生物学、免疫学和RT-PCR鉴定结果,确证MDV N71株为粘膜病病毒,这为深入探讨鹿MD的诊断和防制以及进行分子生物学研究奠定了基础。A deer-origined strain of mucosal disease virus, name N71, was isolated in MDBK cell fecal samples in diarrhea fawn and its properties were identified. The result is that N71 strain exhibited a typical vecuous degeneration, typical virions were observed in the culture fluids with negative staining electron microscope, TCID_(50) was 10^(-5.4) in MDBK cell. N71 strain is sensitive to chloroform、ether and trypsin. The N71 is inactivated after 70 minutes at 56℃, stable at pH7～9, inativated at pH3, and can not agglutinate the red blood cell of sheep、pig、rabbit and chicken. With immunofluorescence antibody of strain 184, fluorescence was observed in MDBK cell infected with N71 strain. A fragment of 400 bp, was amplified from MDBK cell incubated with the isolated virus strain by reverse transcripase-polymerase chain reaction(RT-PCR). From the results described above, it can be concluded that the strain N71is a new strain of deer-origined MDV.

关 键 词：鹿 粘膜病病毒 分离鉴定 

分 类 号：S852.65[农业科学—基础兽医学]