中文摘要：

目的 获得衣原体噬菌体φCPG1的Vp1蛋白,制备抗Vp1的单克隆抗体并检测临床分离的沙眼衣原体标本中是否存在噬菌体.方法 原核表达并纯化衣原体噬菌体φCPG1的衣壳蛋白Vp1,通过杂交瘤技术获得单克隆抗体杂交瘤分泌株,利用ELISA、Western印迹等方法对单克隆抗体进行鉴定,采用动物体内诱生腹水的方法大量制备单克隆抗体并通过G蛋白亲和层析法纯化.采用免疫荧光法检测临床沙眼衣原体噬菌体.结果 获得纯化的Vp1蛋白和3株稳定分泌单抗的杂交瘤细胞株.杂交瘤细胞染色体分析发现染色体数目平均为96,结构上多数为端着丝点染色体,少数亚中部着丝点染色体.3株细胞分泌的单克隆抗体的免疫球蛋白类别均为IgG1.纯化后单克隆抗体效价可达1：102400.利用得到的单克隆抗体检测20份临床标本,结果未发现衣原体噬菌体.结论 成功获得重组的衣原体噬菌体φCPG1Vp1蛋白及抗Vp1蛋白的单克隆抗体,免疫荧光法检测临床沙眼衣原体株中噬菌体未发现阳性标本.

英文摘要：

Objective To express recombinant Vp1 protein of chlamydiaphage φCPG1, prepare monoclonal antibody against Vp1 protein and utilize it to screen clinical isolates of Chlamydia trachomatis. Methods The Vp1 protein was obtained by prokaryotic expression, and monoclonal antibody against this protein was prepared by hybridoma technique. ELISA and Western blot were used to identify monoclonal antibodies. Then,the monoclonal antibody was prepared in quantity by injecting hybridoma cells into the abdominal cavity of BALB/C mice, and purified by using protein G affinity chromatography. Clinical isolates of Chlamydia trachomatis were screened for the chlamydiaphage by immumofluorescence assay using the prepared monoclonal antibody.Results Purified Vp1 protein was obtained. The monoclonal antibody against Vp1 protein was gained after 3times of sub-clone and consistently identified as IgG1. Three hybridoma cell strains that stably secreted monoclonal antibody were generated. Chromosome analysis of hybridoma cells showed that the mean number of chromosome was 96, most of them were telocentric and a few were submetacentric. The titer of purified monoclonal antibody was more than 1： 12 800. Twenty clinical isolates were screened by using the monoclonal antibody and no positive results were obtained. Conclusions The monoclonal antibody against Vp1 protein of chlamydiaphage φCPG1 is successfully prepared, while no chlamydiaphage is detected by immumofluorescence assay using the prepared antibody in 20 clinical isolates of Ct.