何浩,杨造伦,陈云华,李永珍,朱昌福,张式其,杨世庄,彭超德.流行性出血热病原学研究:用A-549细胞从疫区黑线姬鼠肺单层细胞中分离出血热病毒[J].Chinese journal of Epidemiology,1983,4(3):148-152 |
流行性出血热病原学研究:用A-549细胞从疫区黑线姬鼠肺单层细胞中分离出血热病毒 |
Studies on the Etiology of Epidemic Hemorrhagic Fever |
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Abstract: |
用疫区黑线姬鼠肺制备单层细胞,经KHF阳性血清作IFA法检查,将10份KHF荧光阳性(卅以上)的单层细胞培养物接种A-549细胞,发现5株可以在A-549细胞中连续传代的EHF相关因子。以其中ALC96株感染的A-549细胞制作抗原片,检查125份血清,67份EHF患者恢复期血清中66份免疫荧光阳性(98.5%),58份正常人及其它疾病病人血清全部阴性。10例EHF患者双份血清抗体滴度均有4倍以上升高。与呼肠孤Ⅰ、Ⅱ、Ⅲ型及类环状病毒抗血清免疫荧光阴性,与EHF病毒A1、A5,A9株(A-549细胞适应株)抗血清为阳性,其感染性可被EHF病人恢复期血清中和,证明此相关因子是EHF病毒。病毒可通过450nm孔径滤膜。对5-碘脱氧尿嘧啶核苷不敏感,不耐酸(pH≤3.0),对脂溶剂敏感,这些与KHF病毒相似。 |
English Abstract: |
The lung monolayer cell cultures prepared with apodemus agrarius from Epidemic hemorrhagic fever (EHF) endemic area were examined by indirect immunofluorescent antibody (IFA) technique.The sera used cum IFA against KHF virus come from EHF convalescents. A-549 cells were inoculated with 10cultures in which IFA against KHF are positive. Five transmissible strains of EHF-related agent were obtained. Among them a strain ALC96 was identified and its antigenic specifity was confirmed by the following: Sera of 125 subjects were tested by IFA technique with spot-slides prepared from A-549 cells infected by ALC96. 66 out of 67 serum samples of EHF convalescents are positive (98.5%), 58 serum samples of healthy adults and non-EHF patients are negative, ten paired sera from patients with EHF showed more than four times rise in IFA titers. The possibilities of other virus infection such as infection of reovirus (I、II、III type) or orbivirus-like agent can be ruled out by negative results of its corresponding antiserum. At the same time, the spot-slides acted with antiserum against EHF virus A1 A5 A9 (adapted A-549 cell strain) gave positive results.The experiments mentioned above confirmed that the ALC96 agent was EHF virus. The EHF virus is sensitive to lipid-solvents and acid (pH≤3.0), but resistant to idoxyuridene. It is able to pass through bacterial filter with a limiting pore diameter of 450nm.These properties are similar to KHF virus. |
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