抗体制备ELISA试剂诊断肾综合征出血热  

ELISA reagent for detecting hemorrhagic fever with renal syndrome by using antibody of Hantaan virus

作　　者：王敬军[1] 余鹏博[1] 董建华[1] 张家驹[1] 邢爱华[1] 王百锁[1] 李翔[1] 田小平[1] 

机构地区：[1]陕西省疾病预防控制中心,西安710054

出　　处：《中国公共卫生》2005年第12期1425-1426,共2页Chinese Journal of Public Health

基　　金：陕西省首届医药卫生重点科研基金项目(99zh-001)

摘　　要：目的研究汉坦病毒重组抗原及其单克隆抗体制备双抗夹心酶联免疫吸附(ELISA)试剂早期诊断肾综合征出血热(HFRS)。方法将汉坦病毒76 118株S 1.3 kb基因克隆于pET 28 a质粒,诱导大肠埃希菌表达出48 kd的汉坦病毒核蛋白抗原。以此抗原免疫Balb/c小白鼠,取其B淋巴细胞与骨髓瘤细胞融合,筛选阳性杂交瘤细胞株,小鼠腹腔接种,制备单克隆抗体。用重组抗原及其单克隆抗体,制备ELISA诊断试剂,并与美国产汉坦病毒重组抗原ELISA诊断试剂和间接免疫荧光法(IFA)比较。结果自产重组抗原及其单克隆抗体制备的双抗夹心ELISA试剂与美国产抗原ELISA试剂在87例HFRS和56例非HFRS诊断中完全吻合。结论自产ELISA试剂可用于早期诊断HFRS,且意义重大。Objective To detect Hemorrhagic Fever with Renal Syndrome（HFRS） earlier and sensitively, the recombinant nucleocapsid protein（rNP）and monoclonal antibody（ McAb）of Hantaan virus were used and ELISA reagent was allocated. Methods The genome S 1.3 kb of Hantaan virus 76 118 was transfered to plasmid of E. coli to express nucleocapsid protein. Lymphocytes B were taken from mice which were immunized by the rNP. The sensitized lymphocytes B and myeloma cells were fused together for getting the McAb of rNP, and IgM capturing ELISA was established with rNP and McAb. Our ELISA reagent was compared with IFA and ELISA reagent which was produced with American rNP and McAb. Results The sensitivity and specificity of our ELISA reagent met the standard of the ELISA reagent with American rNP and McAb completely. Conclusion It is great meaningful for using our IgM capturing ELISA to detect HFRS.

关 键 词：肾综合征出血热(HFRS) 重组核蛋白 单克隆抗体 酶联免疫试验 

分 类 号：R392-33[医药卫生—免疫学]