中文摘要：

目的采用二维电泳、免疫印迹法和质谱技术鉴定日本血吸虫可溶性虫卵抗原（SEA）成分中具有早期诊断价值的抗原分子。方法通过等电聚焦以及十二烷基磺酸钠聚丙烯酰胺凝胶电泳对SEA进行二维电泳,将胶上蛋白质斑点转移到硝酸纤维素膜上,再分别与日本血吸虫感染前、后不同时间点的小鼠血清反应,以辣根过氧化物酶标记的羊抗鼠IgG作为二抗,进行免疫印迹试验,寻找SEA中能与血吸虫感染早期血清反应的蛋白斑点。通过高效液相色谱串联质谱（LC MS/MS）分析鉴定其蛋白成分。结果对血吸虫感染1、2、6周以及未感染健康小鼠血清识别SEA的二维电泳免疫印迹图谱进行匹配分析发现,SEA中有2个蛋白斑点能被血吸虫感染后1、2、6周小鼠血清识别,另有3个蛋白斑点仅被血吸虫感染后6周小鼠血清识别。经LC MS/MS鉴定,2个被血吸虫感染早期血清识别的SEA蛋白分子分别为热休克蛋白（HSP70）和葡萄糖调节蛋白（Grp78/Bip）。结论 SEA中HSP70和Grp78/Bip可能具有血吸虫感染早期诊断价值。

英文摘要：

Objective To identify the molecules of soluble egg antigen（SEA） for early diagnosis of Schistosoma japonicum infection by two-dimensional electrophoresis（2-DE）,immunoblotting and liquid chromatography with tandem mass spectrometry（LC-MS/MS）.Methods The 2-DE of SEA was executed through first direction isoelectric focusing（IEF） in immobilized pH gradient gel 3-10（IPG3-10） and second direction SDS-PAGE.The protein dots of SEA on the SDS-PAGE gel were transferred to nitrocellulose membrane.These nitrocellulose membranes were responded to the sera of healthy,sera of mice at 1 week,2 weeks and 6 weeks post-infection respectively,then the membrane color was developed with the second antibody of HRP labeled goat anti-mouse IgG conjugate and substrate DAB.The protein dots recognized by sera of mice in the early stage of schistosome infection were identified by LC-MS/MS.Results After matching and analyzing the Western blot patterns of SEA responding to acute infection sera（1 week and 2 weeks post-infection）,chronic infection sera（6 weeks post-infection） and healthy sera by PDQuest 1.0 software,two protein dots were found to be recognized by sera of mice at 1 week,2 weeks and 6 weeks post-infection,and three protein dots were only recognized by the sera of mice at 6 weeks post-infection,no protein dot was recognized by healthy mouse sera.The data of LC-MS/MS showed that the two protein molecules recognized by the sera of mice with schistosome infection in the early stage were heat shock protein 70（HSP70） and 78 kDa glucose-regulated protein（Grp78/Bip） respectively.Conclusion The results of this study preliminarily indicate that HSP70 and Grp78 in SEA have early diagnostic value for S.japonicum infection.