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
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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。Abstract:Objective To detect Hemorr hagic 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 nucleo capsid protein.Lymphocytes B were taken from mice which were immunized by the rNP.The sensitized lymphocytes Band 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 IFAand ELISA reagent which was produced with American rNP and McAb.Results The sensitivity and specificity of our ELISA reagent met the standar d of the ELISA reagent with American rNP and McAb completely.Conclusion It is great meaningful for using our IgM capturing ELISA to detect HFRS.
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