流行性出血热病毒Vero-E6细胞病变株的分离和鉴定

从EHF患者血清中分离到8株EHF病毒,在E 6细胞中传代适应过程中,发现有5株EHF病毒分别在第4～10代出现规律的细胞病变(CPE),对其中两个CPE株(HFN-04和-19)进行了一系列特异性鉴定试验。实验结果表明:(1)病毒的CPE发展与病毒增殖动态完全一致;(2)CPE株的主要生物学和理化性状均符合EHF病毒;(3)用CPE株病毒感染细胞抗原片检测EHF患者双份血清,其恢复期血清抗体均呈4倍以上增高反应;(4)病毒的CPE可被患者血清及EHF、KHF免疫血清所中和,不能被正常兔血清和Reo病毒1—3型多价血清所中和;(5)CPE株病毒感染的E_6细胞几乎能完全吸收EHF患者血清及EHF、KHF抗血清中的特异性抗体;(6)CPE株病毒感染鼠脑悬液与EHF病毒A9株McAb25-1致敏血球呈阳性血凝反应。据此,首次证实了EHF病毒至少有部分毒株能引起E_6细胞规律性CPE。EHF病毒CPE株的发现为EHF的诊断和防治工作提供了有利条件。

Isolation and Identification of cytopathogenic Effect Strains of Epidemic Hemorrhagic Virus in Vero-E_6 Cell Culture

Five strains out of the 8 of epidermic hemorrhagic fever(EHF)virus isolated from the serum of the acute phase of EHF patients in Nanchong, Sichuan were found to have cytopathogenic effect(CPE)through 4th to 10th passages in Vero-E6 cell culture. The biological, physico-chemical and antigenic characteristics of HFN-04 and HFN-19, 2 out of the 5 CPE strains, were identified. The results were as follows;( 1 ) The CPE development and the dynamic proliferation of the virus areconsistent. ( 2 ) The main biological and physico-chemical characteristics of the 2 CPEstrains arc the same as those of EHF virus.( 3 ) When paired samples of EHF patients' serum were tested with the slide antigens of E6 cells infected with these CPE strains, all the convalescent serum demonstrated an increase by 4 fold or more of the IFA. titer.( 4 ) The CPE of the virus can be neutralized by EHF patient's serum or EHF and KHF antisera, but not by normal rabbit serum or the multivalent serum of Reovirus type 1-3.( 5 ) The E6 cells infected by CPE strains of virus can almost fully absorbthe specific antibodies in EHF patients or in EHF and KHF antisera.( 6 ) The suspension of the mouse brain infected by CPE strains can formpositive agglutination reaction with the sentitized blood cells by EHFvirus strain A9 McAb 25-1.On the basis of our observation, it can be concluded that at least certain strains of EHF virus can produce CPE in Vero-E6 cell culture.