阴道毛滴虫全虫抗原的制备及其免疫原性鉴定  

Preparation of an antigen found in Trichomonas vaginalis trophozoites and detection of its immunogenicity

作　　者：许少莉 杨森[1] 王黎[1] 薛长贵[1] 

机构地区：[1]郑州大学基础医学院人体寄生虫学教研室,河南郑州450001

出　　处：《中国病原生物学杂志》2014年第11期1016-1019,共4页Journal of Pathogen Biology

摘　　要：目的制备阴道毛滴虫全虫抗原并用其免疫家兔,观察其免疫原性。方法将阴道毛滴虫临床分离株接种于TYM培养基进行连续培养,达到纯培养后制成全虫抗原,经背部皮下多点注射免疫家兔,间隔2周加强免疫（共2次）,末次免疫后7d采血,分离血清,采用ELISA检测抗体效价,达标后采集、分离并纯化免疫血清,采用Western blot和间接免疫荧光法检测其免疫原性。结果阴道毛滴虫在连续培养3代后即达到纯培养,SDS-PAGE显示提取的全虫抗原蛋白带分布在10-150ku之间,其中30-70ku之间的蛋白带最为密集。ELISA检测显示阴道毛滴虫全虫抗原注射家兔后引起较强的免疫反应,产生的抗体滴度〉1∶1×10^6;Western blot检测共有15种全虫抗原蛋白组分可被免疫兔血清识别,免疫荧光染色显示阴道毛滴虫虫体可与全虫抗原免疫兔血清发生免疫反应,呈现红色荧光。结论阴道毛滴虫全虫抗原具有良好的免疫原性,用该抗原免疫家兔可获得较高滴度的抗血清。Objective To prepare an antigen found in Trichomonas vaginalis trophozoites and to immunize rabbits with the trophozoite antigen in order to detect its immunogenicity. Methods Clinical isolates of T.vaginalis were seeded in TYM medium and maintained as axenic cultures to produce the trophozoite antigen.Rabbits were immunized by subcutaneous injections of the trophozoite antigen in the skin of the back.Rabbits were either given a booster（2weeks later）or they were not immunized.Seven days later,blood was collected,serum was separated,and the antibody titer was detected with ELISA.Once the antibody titer met criteria,the serum was separated and purified,and immunogenicity was determined using Western blotting and IFA. Results Three passages yielded a pure culture of T.vaginalis.SDS-PAGE results indicated that the protein bands for the trophozoite antigen were between 10-150 ku,and the most intense band was between 30-70 ku.ELISA results indicated that the trophozoite antigen caused a strong immunoreaction in immunized rabbits;the antibody titer was greater than 1：1×10^6.Western blotting results indicated that 15 antigen proteins were recognized by serum from immunized rabbits;only 12 proteins were recognized by mouse serum.Immunofluorescence results indicated that the immunoreaction between the trophozoite antigen and serum led to red fluorescence. ConclusionImmunizing rats with an antigen found in Trichomonas vaginalis trophozoites resulted in a high antibody titer,indicating that the antigen has good immunogenicity.

关 键 词：阴道毛滴虫 抗原 ELISA 免疫 

分 类 号：R382.21[医药卫生—医学寄生虫学]