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机构地区:[1]中国科学院武汉病毒研究所
出 处:《畜牧兽医学报》1999年第4期348-352,共5页ACTA VETERINARIA ET ZOOTECHNICA SINICA
摘 要:采用氯仿、聚乙二醇一硫酸葡聚糖钠盐二相系统和蔗糖密度梯度离心法,从患兔出血症死亡兔肝组织中提取、纯化病毒。该病毒粒子无囊膜,呈20面体对称,直径一般为33~37nm。其三角剖分数T=3,共有32个子粒,每一子粒为中空、外径为9nm左右的圆形轮廓。经蔗糖密度梯度离心后,可获得沉降系数为166s的病毒颗粒,这种颗粒具有很强的感染性。二苯胺试验,吖啶橙染色、甲醛试验及热变性曲线表明,该病毒是单链DNA病毒。电镜下经甲酰胺法展层的病毒核酸呈单链线状,分子量平均为21×106道尔顿。53s中空的圆形子粒,有很强的血凝性,而对照组铁蛋白和同一条件下所制备正常兔肝成份无凝血特性,说明53s的9nm粒子应为病毒外壳蛋白子粒。该病毒粒子经琼脂糖双扩散试验显示不同于兔细小病毒标准抗血清产生沉淀线。根据试验结果显示,我们认为该RHDV可能是兔出血症病毒的新血清型。In this paper a strain of Rabbit Hemorrhagic Disease Virus(RHDV)was isolated and purified from the diseased rabbit livers with a method of using chloroform,two phase of polyethylene glycol dextran sulfate sodium and sucrose density gradient centrifugation.Purified virus was nonenveloped,icosahedral symmetry with a triangulation number of 3,and 33~37nm in diameter.The capsid was composed of 32 capsomeres with central holes in an outer diameter of about 9nm.Two types of viral partcles having different sedimentation coefficient,130s and 166s could be identified after sucrose density gradient centrifugation.Viral nucleic acid was extracted from purified virus by using SDS proteinase K phenol.Tests with diphenylamine,formaldehyde,and staining with acridine orange as well as the curves of thermal denaturation showed that this strain of virus had a single stranded DNA.The molecular weight of the ssDNA was approx 2 1×10 6 dalton as determined by electron microscopy.Data indicate RHDV cant react with antiserum of standard Parvovirus of Rabbit and so on,So we suggest the isolate of RHDV is a new viral strain.
分 类 号:S858.291[农业科学—临床兽医学] S852.65[农业科学—兽医学]
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