抗Trx单克隆抗体用于血吸虫病诊断的研究  

Application of Double Antibody Sandwich ELISA for Detection of Trx of Schistosoma Japonicum

作　　者：吴玉龙[1,2] 程梅[3] 刘春会[1] 耿丽[1] 张珍[1] 杜镇镇[1] 李波清[1] 李雍龙[4] 

机构地区：[1]滨州医学院病原生物学教研室,山东烟台264003 [2]南京医科大学卫生部抗体技术重点实验室,江苏南京210029 [3]滨州医学院护理学院,山东滨州256603 [4]华中科技大学基础医学院人体寄生虫学教研室,湖北武汉430030

出　　处：《现代生物医学进展》2014年第6期1057-1061,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81072429)

摘　　要：目的:建立双抗体夹心ELISA法检测日本血吸虫硫氧还蛋白(Thioredoxin,Trx)。方法:用重组日本血吸虫Trx(rTrx)蛋白免疫BALB/c小鼠,筛选高滴度、高特异性的单克隆抗体建立双抗体夹心ELISA法。通过检测日本血吸虫排泄-分泌物(excretorysecretions,ES)与rTrx的浓度评价该方法的敏感性;通过对健康人血清的检测确定其特异性;通过对布氏姜片吸虫病、华支睾吸虫病、卫氏并殖吸虫病、囊虫病患者血清进行交叉反应试验,评价该方法的特异性。结果:获得2株稳定分泌抗rTrx蛋白单克隆抗体的杂交瘤细胞株,命名为McTrx1和McTrx2。以McTrx1为包被抗体,HRP-McTrx2为酶标抗体,建立的双抗体夹心ELISA可检测出ES的最低浓度为4.8μg/ml,检测出rTrx的最低浓度为1.2μg/ml。该方法的特异性为96%。结论:以抗rTrx蛋白单克隆抗体McTrx1与McTrx2为基础建立的双抗体夹心ELISA法具有较高的特异性。Objective： To establish a double antibody sandwich ELISA method for detection of Trx of Schistosoma japonicum. Methods： BALB/c mice were immunized with recombinant Trx （rTrx） of Schistosoma japonicum. The hybridomas that secreted high titer of monoclonal antibodies （mAbs） with high specificity were screened and used to establish the double antibody sandwich ELISA for the detection of rTrx. In order to evaluate the sensitivity of the method, the concentration of ES antigen and rTrx was detected, respectively. Serum samples from healthy people and patients with fasciolopsiasis, clonorchiasis,paragonimiasis, cysticercosis were examined by the same method to estimate the specificity. Results： Two hybridoma cell lines, McTrxl and McTrx2, were developed for secreting mAbs against Trx. The McTrxl was used as coating antibody, and HRP-labeled McTrx2 as secondary antibody. The double antibody sandwich ELISA detecting Trx was developed with a minimum concentration of 4.8 p^g/mL for ES of Schistosoma japonicum and 1.2 ixg/mL for Trx. An overall specificity of 96 % was determined. Conclusion： The double antibody sandwich ELISA based on McTrxl as coating antibody and McTrx2 as secondary antibody has a high specificity.

关 键 词：日本血吸虫 单克隆抗体 硫氧还蛋白 双抗夹心ELISA 

分 类 号：R383.2[医药卫生—医学寄生虫学]