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应用细胞培养技术和免疫荧光(IF)检查方法,从汉中地区褐家鼠分离出EHF 病毒R_6、R_(10)株。分离的病毒能在鼠体和细胞两个不同宿主系统连续稳定传代;检查10份确诊的EHF 病人双份血清,抗体滴度均有4倍以上增高;病毒抗原能与EHF病人阳性血清、兔抗KHFV76-118株免疫血清及多个单克隆抗体(M_cAb)反应,而不能与正常人和非出血热病人血清反应;兔抗KHFV76-118株免疫血清能阻断病毒株抗原同EHFM_cAb4E_7的阳性反应,抗呼肠弧病毒Ⅰ～Ⅲ免疫血清、抗类环状病毒免疫血清、抗CMV 免疫血清和抗HSV 免疫血清均无此阻断作用;用感染的VeroE_(-6)细胞作超薄切片,电镜下观察到大小约100nm 典型病毒颗粒;M_cAb 型别鉴定试验确定为EHFV野鼠型毒株。
EHF virus R_6 and R_ (10) strains were isolated from the brown spot mice of Hanzhong area by using cell culture technique and immunofluorescence (IF) method. The isolated virus can be continuously and serially passaged in two different host systems of mice and cells. The double serum of 10 patients diagnosed with EHF was tested, and the antibody titers increased more than 4 times. The virus antigens were able to react with the positive serum and rabbit anti-EHF KHFV76-118 immune serum and multiple monoclonal antibodies (M_cAb) reaction, but not normal and non-hemorrhagic fever serum; Rabbit anti-KHFV76-118 immune serum can block the virus strain antigen with EHFM_cAb4E_7 positive reaction, Anti-reovirus serogroup Ⅰ ~ Ⅲ immune serum, anti-class cyclic virus immune serum, anti-CMV immune serum and anti-HSV immune serum were not blocked. VeroE_ (- 6) The typical size of about 100nm virus particles were observed; M_cAb type identification test identified as EHFV wild-type strain.