目的制备肺炎支原体(Mycoplasma pneumoniae,Mp)重组P1蛋白多克隆抗体,为Mp抗原诊断试剂的制备奠定基础。方法用Mp重组P1蛋白免疫家兔,ELISA法测定免疫血清抗体效价;硫酸铵沉淀及离子交换层析法纯化抗体;紫外分光光度法测定抗体的浓度;SDS-PAGE分析抗体的纯度;量子点标记纯化抗体,分析抗体的免疫活性;直接免疫荧光法检测抗体的特异性。结果重组P1蛋白兔免疫血清抗体效价为1∶25 600;纯化的抗体浓度为3.689μg/μl;SDS-PAGE显示相对分子质量58 000的重链条带;用量子点标记纯化的抗体仍保持较高的免疫活性,与其他呼吸道常见病原菌均未发生特异性结合。结论获得的纯化Mp P1蛋白IgG抗体具有较高的免疫活性和特异性,可用于临床Mp感染的抗原诊断。 Objective To prepare a polyclonal antibody against recombinant P1 protein of Mycoplasma pneumoniae (Mp), and lay a foundation for the preparation of Mp antigen diagnostic reagent. Methods Rabbit was immunized with Mp recombinant P1 protein, the titer of immune serum was determined by ELISA, the antibody was purified by ammonium sulfate precipitation and ion exchange chromatography, the concentration of antibody was determined by ultraviolet spectrophotometry, the purity of antibody was analyzed by SDS-PAGE, The purified antibody was labeled to analyze the immunological activity of the antibody. The specificity of the antibody was detected by direct immunofluorescence. Results The antibody titer of recombinant P1 protein was 1:25 600. The concentration of purified antibody was 3.689 μg / μl. SDS-PAGE showed a heavy chain with molecular weight of 58 000. High immune activity, and other respiratory pathogens are not specific binding. Conclusion The purified IgG antibodies against Mp P1 protein have high immunogenicity and specificity and can be used to diagnose antigen in clinical Mp infection.