流行性出血热病毒单克隆抗体的制备及其IgM捕获ELISA的建立  

Preparation of Antibodies against EHF Virus and Its Application in Diagnosis

作　　者：王美亮[1] 陈淑清 袁湘叶 彭静[2] 周学良[2] 郑海金[3] 

机构地区：[1]解放军473医院检验科,甘肃兰州730070 [2]卫生部兰州生物制品研究所,甘肃兰州730046 [3]兰州大学生命科学学院,甘肃兰州730000

出　　处：《兰州大学学报（自然科学版）》1999年第4期94-97,共4页Journal of Lanzhou University(Natural Sciences)

摘　　要：用灭活的流行性出血热病毒（ＥＨＦＶ） 免疫小鼠获得４ 株能分泌ＥＨＦＶ 特异性单克隆抗体（ＭｃＡｂ） 的杂交瘤细胞．Ｗｅｓｔｅｒｎ ｂｌｏｔ 结果表明４ 株ＭｃＡｂ 能专一地识别ＥＨＦ 病毒的分子量为５６ｋＤ的囊膜糖蛋白Ｇ２ ．交叉实验结果表明４ 株ＭｃＡｂ 与野鼠型病毒株７６１１８ 和家鼠型病毒株Ｌ９９ ，Ｒ２２ 等均有反应，而与流感病毒、轮状病毒等均无交叉反应，表明４ 株ＭｃＡｂ 具有较高的特异性．用６Ｂ１１ ＭｃＡｂ 组建ＩｇＭ 抗体捕获ＥＬＩＳＡ 试剂盒，试测临床标本．２５ 例临床确诊ＥＨＦ病人血清中特异性ＩｇＭ 抗体全部阳性．７０ 例正常献血员血清全部阴性．ＥＬＩＳＡ 试验方法简单，敏感性好，可建立试剂盒应用于临床．Cell fusion was performed by using Sp2/O myeloma cells and spleen cells from BALB/c mice immunized with EHFV antigen. Four hybrid cell lines secreting monoclonal antibodies(McAb) against EHF virus were obtained. The results of westernblot proved that they all recognized the 56KD membrane glucoprotein G 2 of EHF virus. McAbs can react with EHFV 76 118, L 99 and R 22 strains, but it cant with influenza virus and rotavirus as contrast. The IgM antibody capture ELISA(MacELISA) test for IgM antibodies against EHFV was established. Serum specimens collected from 25 EHF patients were all tested to be positive while 70 normal human serum specimens were all negative. The results demonstrated that MacELISA was more sensitive than IFAT and can be applied in clinical diagnosis.

关 键 词：流行性出血热 单克隆抗体 IGM ELISA EHFV 

分 类 号：R373.32[医药卫生—病原生物学] R512.8[医药卫生—基础医学]