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目的制备抗人3型副流感病毒核衣壳蛋白特异性单克隆抗体,为研发3型副流感病毒新型诊断试剂提供基础。方法利用重组克隆表达技术,获得可溶性的重组核衣壳蛋白;采用纯化的重组核衣壳蛋白免疫小鼠,采用杂交瘤技术制备抗人3型副流感病毒核衣壳单克隆抗体;采用ELISA及间接免疫荧光法筛选并鉴定可识别天然3型副流感病毒核衣壳蛋白单克隆抗体。结果获得9株分泌抗重组核衣壳蛋白的特异性单克隆抗体的细胞株,其中2株杂交瘤细胞分泌的单克隆抗体能识别天然的3型副流感病毒核衣壳蛋白。结论利用重组核衣壳蛋白作为免疫原制备出了识别天然核衣壳蛋白的单克隆抗体,并且具有较高的反应性。
Objective To prepare specific monoclonal antibodies against human parainfluenza virus type 3 nucleocapsid protein and provide a basis for the development of novel diagnostic reagents for type 3 parainfluenza virus. Methods The purified recombinant nucleocapsid protein was obtained by recombinant cloning technique. The purified recombinant nucleocapsid protein was used to immunize mice and the monoclonal antibody against human parainfluenza virus type 3 nucleocapsid was prepared by hybridoma technique. Indirect immunofluorescence screening and identification of monoclonal antibodies that recognize nucleocapsid protein of the native parainfluenza type 3 virus. Results Nine cell lines secreting specific monoclonal antibodies against the recombinant nucleocapsid protein were obtained. The monoclonal antibodies secreted by the two hybridoma cells could recognize the native type 3 parainfluenza virus nucleocapsid protein. Conclusion The recombinant nucleocapsid protein was used as immunogen to prepare a monoclonal antibody that recognizes natural nucleocapsid protein and has high reactivity.