抗柔嫩艾美耳球虫EtRP单克隆抗体的制备及其免疫保护效果  

Preparation of Monoclonal Antibody Against Eimeria tenella and Its Immune Protection for Chicken

作　　者：陈浩[1] 蔡宇威 刘兰[1] 梁雄燕[1] 卢渊录 

机构地区：[1]长江大学动物科学学院,湖北荆州434025

出　　处：《中国家禽》2017年第7期20-23,共4页China Poultry

摘　　要：棒状体蛋白(EtRP)在柔嫩艾美耳球虫入侵宿主细胞的过程中起重要作用。该蛋白在球虫发育阶段的早期表达,具有保护性抗原的性质,其相应抗体可抑制球虫对宿主的感染。采用纯化后的p GEX-6P-1-EtRP重组蛋白免疫BALB/c小鼠制备了抗EtRP的单克隆抗体,将不同剂量的单克隆抗体注射试验鸡群再对免疫鸡群攻虫,7 d后致死鸡群并计算卵囊减少率、相对增重率以及盲肠病变评分情况。结果表明,所制备的单克隆抗体亚型为IgG1,经Western-blot检测能特异性识别重组蛋白,经间接ELISA测定其效价大于1∶128 000。此外,单克隆抗体对鸡的被动保护性试验结果显示相对增重率、病变评分、卵囊减少率均随着免疫剂量的增加而增大,表明一定剂量的单克隆抗体可以有效减少柔嫩艾美耳球虫对鸡群的感染。Rhoptry protein （EtRP）plays an important role in the process of Eimeria invading host cell. The protein is expressed at the early developmental stages of Eimeria,it has the nature of protective antigen,the antibody could inhibit the infection of the host. In this experiment the monoclonal antibody against EtRP was obtained by immunizing BALB/c mice with purified recombinant pGEX-6P-1-EtRP protein. Subsequently,different doses of monuclonal antibody was injected into the experimental group of chicken,then Eimeria tenella was injected into immunized chickens. The chicken oocysts reduction rate, relative growth rate and cecal lesion scores were detected after 7 d. The results showed that the monoclonal antibody could specifically recognize the recombinant protein by Western-blot,and the titer was more than 1：128 000 by indirect ELISA and the subtype was identified as IgGl by the immunoglobulin detection kit. In addition, the results of passive protection test showed that the relative weight gain rate, lesion score and egg reduction rate were all increased with the increase of immune dose. It was proved that a certain dose of monoclonal antibody could effectively reduce the infection of Eimeria tenella to chickens.

关 键 词：柔嫩艾美耳球虫 棒状体蛋白(EtRP) 单克隆抗体 免疫保护 

分 类 号：S855.9[农业科学—临床兽医学]