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目的以亮氨酸氨肽酶(LAP)作为检测抗原,观察日本血吸虫感染后不同时期动物血清中相应抗体的动态变化特征。方法应用免疫印迹试验(Western blot)鉴定日本血吸虫LAP(SjLAP)重组蛋白的表达,并利用His镍柱纯化目的蛋白。采用酶联免疫吸附试验(ELISA)检测感染前、后不同时期小鼠及猪血清中LAP诱导抗体产生的变化特征。结果重组蛋白SjLAP可与anti-His单克隆抗体反应,并可被日本血吸虫感染后小鼠血清所识别。获得高浓度重组蛋白SjLAP,以该蛋白为抗原,采用ELISA法检测发现感染后4周猪血清和感染后6周小鼠血清抗体水平达到高峰;且随着虫卵大量出现,抗体水平均显著下降(P均<0.01)。结论 SjLAP在感染早期的高表达可能对日本血吸虫病早期诊断具有一定的参考价值。
Objective To detect the dynamic changes of the corresponding antibodies in serum of different sera of Schistosoma japonicum infected with leucine aminopeptidase (LAP) as antigen. Methods Western blotting was used to identify the expression of SjLAP recombinant protein of Schistosoma japonicum and His Ni column was used to purify the target protein. Enzyme-linked immunosorbent assay (ELISA) was used to detect the changes of LAP-induced antibody production in mouse and pig serum at different stages before and after infection. Results The recombinant protein SjLAP reacted with anti-His monoclonal antibody and was recognized by the sera of mice infected with Schistosoma japonicum. The recombinant protein SjLAP with high concentration was obtained. Using this protein as antigen, the level of serum antibody in pig serum at 4 weeks after infection and at 6 weeks after infection reached the peak by ELISA, and the level of antibody was significantly decreased with the appearance of large numbers of eggs (P <0.01). Conclusion The high expression of SjLAP in the early stage of infection may have some reference value for the early diagnosis of schistosomiasis japonica.