| 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.抗 Trx 单克隆抗体用于血吸虫病诊断的研究 *[J].,2014,14(6):1057-1061 |
| 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* |
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| DOI： |
| 中文关键词: 日本血吸虫 单克隆抗体 硫氧还蛋白 双抗夹心 ELISA |
| 英文关键词: Schistosoma Japonicum,Monoclonal antibody Trx Double antibody sandwich ELISA |
| 基金项目:国家自然科学基金项目 （81072429 ） |
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| 中文摘要: |
| 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 4.8 μg/ml， 检测出 rTrx 的最低浓度为 1.2 μg/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。 |
| 英文摘要: |
| ABSTRACT 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, McTrx1 and McTrx2, were developed for
secreting mAbs against Trx. The McTrx1 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 μg/mL for ES of
and 1.2 μg/mL for Trx. An overall specificity of 96 % was determined.Conclusion: The double antibody sandwich ELISA based on
McTrx1 as coating antibody and McTrx2 as secondary antibody has a high specificity |
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