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将ＰＣＲ扩增的ＨＴＶ７６－１１８ＲＮＡ小片段的ｃＤＮＡ克隆插入到载体ｐＤＳ５６／ＲＢＳⅡ－（Ｏ）－６Ｈｉｓ中，通过ＩＰＴＧ诱导表达以及镍螯合层析纯化重组ＮＰ，经过ＳＤＳ－ＰＡＧＥ、免疫转印和ＥＬＩＳＡ方法分析其蛋白特性，证实该重组ＮＰ与毒粒ＮＰ相同，均为４９．６ｋｕ，能与肾综合征出血热（ＨＦＲＳ）患者血清产生特异性反应。以此重组ＮＰ作为抗原制备抗ＮＰ单克隆抗体，获得两株持续稳定分泌单克隆抗体的杂交瘤细胞株。用抗ＮＰＭｃＡｂ建立检测ＩｇＭ抗体的ＥＬＩＳＡ捕获法。证实纯化重组ＮＰ作为抗原，避免了组织培养病毒的感染性和非特异性以及难以标准化的缺点。所建立的ＥＬＩＳＡ捕获法检测ＨＦＲＳ患者血清中ＩｇＭ抗体的方法特异性强、敏感性高，可作为ＨＦＲＳ的早期诊断试剂
The cDNA clone of the PCR-amplified HTV76-118RNA fragment was inserted into the vector pDS56 / RBSII- (O) -6His. The recombinant NP was induced by IPTG and purified by nickel chelation chromatography. After SDS-PAGE, immunoprecipitation and The protein characteristics of the recombinant NPs were analyzed by ELISA. The results showed that the recombinant NP was 49.6ku, the same as virion NPs, and could specifically react with the serum of patients with hemorrhagic fever with renal syndrome (HFRS). Using this recombinant NP as an antigen to prepare an anti-NP monoclonal antibody, two hybridoma cell lines that consistently and stably secrete monoclonal antibodies were obtained. An ELISA capture method to detect IgM antibodies was established with anti-NPMcAb. The purified recombinant NPs were confirmed as antigens, avoiding the infectivity and nonspecificity of tissue culture viruses as well as the disadvantages of difficult to standardize. The established ELISA capture method for detecting IgM antibodies in serum of HFRS patients has strong specificity and high sensitivity and can be used as an early diagnosis reagent for HFRS