Wang Jie, Yu Chuan-Xin, Yin Xu-Ren, Qian Chun-Yan, Song Li-Jun, Xu Yong-Liang, He Wei, Cao Guo-Qun
Jiangsu Institute of Parasitic Diseases, Key Laboratory on Technology for Parasitic Disease Prevention and Control, Ministry of Health, Wuxi 214064, China.
Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi. 2011 Jun;23(3):273-8.
To find out the candidate antigen for immunoreagent, which could be used to diagnose Schistosoma japonicum infection early in mice.
The mice were infected with cereariae of S. japonicum Chinese mainland strain. The sera of mice before and after infection at different time were collected. The recombinant fusion protein (GST-HD) of the large hydrophilic domain (HD) of 23 kDa membrane protein of S. japonicum with the Glutathione-S-transferase (GST) of S. japonicum, soluble eggs antigen (SEA), TSP2 hydrophilic domain of S. japonicum (TSP2HD), IL4-inducing principle of S. mansoni eggs (IPSE), fusion protein GST-SjMP10 (SjMP-10), and recombinant S. japonicum (Chinese strain) signaling protein 14-3-3 (Sj14-3-3) were used as diagnostic antigens, the specific IgG and IgM antibodies were measured respectively by enzyme linked immunosorbent assay (ELISA). The antigens with the value of diagnosing schistosomiasis early were screened by analyzing the changes of the levels of specific IgG (or IgM) antibodies and the positive rates of specific antibodies in the sera of mice before and post infection at different time. Moreover, the antigen's value of early diagnosis was further validated by Immunoblot.
On the 18th, 21st and 28th day post infection, the positive rates of specific antibody IgM against GST-HD were 60%, 70% and 100%, respectively; the positive rates of specific antibody IgG against GST-HD were 40%, 60% and 90%, respectively. The positive rates of antibody IgM against SEA were 50%, 60% and 90%, respectively; the positive rates of antibody IgG against SEA were 20%, 50% and 70%, respectively. The positive rates of IgM against TSP2HD were 30%, 40% and 50%, respectively; the rates of IgG against TSP2HD were 20%, 30% and 70%, respectively. The positive rates of IgM against IPSE were 20%, 30% and 50%, respectively; the positive rates of IgG against IPSE were 20%, 30% and 60%, respectively. The positive rates of IgM against SjMP-10 were 10%, 20% and 20%, respectively; the rates of IgG against SjMP-10 were 10%, 20% and 30%, respectively. The positive rates of IgM against Sj14-3-3 were 0, 10% and 20%, respectively; the rates of IgG against Sj14-3-3 were 0, 10% and 30%, respectively. The sensitivities of GST-HD and SEA for diagnosing schistosome infection early in mice were significantly higher than those of Sj14-3-3, IPSE, TSP, and MP-10. The sensitivity of IgM was higher than that of IgG. In Western blotting, the about 73 kDa protein band of SEA was recognized by sera of mice one week post infection and showed stronger reaction as the infected time went on. Moreover, the bands (33 kDa) of GST-HD were earliest recognized by the mouse sera on the 10th day post-infection, the bands showed strong reaction with the mouse sera of 5-week post-infection.
The GST-HD fusion protein and the protein of which molecular weight is about 73 kDa of SEA have the early diagnostic value for schistosomiasis, and the sensitivity of Immunoblot is higher than that of ELISA.
寻找可用于早期诊断小鼠日本血吸虫感染的免疫试剂候选抗原。
用中国大陆株日本血吸虫尾蚴感染小鼠，收集不同时间点感染前后小鼠的血清。以日本血吸虫23 kDa膜蛋白的大亲水结构域（HD）与日本血吸虫谷胱甘肽-S-转移酶（GST）的重组融合蛋白（GST-HD）、可溶性虫卵抗原（SEA）、日本血吸虫TSP2亲水结构域（TSP2HD）、曼氏血吸虫虫卵IL4诱导原（IPSE）、融合蛋白GST-SjMP10（SjMP-10）及重组日本血吸虫（中国株）信号蛋白14-3-3（Sj14-3-3）作为诊断抗原，采用酶联免疫吸附试验（ELISA）分别检测特异性IgG和IgM抗体。通过分析不同时间点感染前后小鼠血清中特异性IgG（或IgM）抗体水平变化及特异性抗体阳性率，筛选出可早期诊断血吸虫病的抗原。此外，通过免疫印迹进一步验证该抗原的早期诊断价值。
感染后第18、21和28天，GST-HD特异性抗体IgM阳性率分别为60%、70%和100%；GST-HD特异性抗体IgG阳性率分别为40%、60%和90%。SEA抗体IgM阳性率分别为50%、60%和90%；SEA抗体IgG阳性率分别为20%、50%和70%。TSP2HD IgM阳性率分别为30%、40%和50%；TSP2HD IgG阳性率分别为20%、30%和70%。IPSE IgM阳性率分别为20%、30%和50%；IPSE IgG阳性率分别为20%、30%和60%。SjMP-10 IgM阳性率分别为10%、20%和20%；SjMP-10 IgG阳性率分别为10%、20%和30%。Sj14-3-3 IgM阳性率分别为0、10%和20%；Sj14-3-3 IgG阳性率分别为0、10%和30%。GST-HD和SEA早期诊断小鼠血吸虫感染的敏感性显著高于SjMP-10、IPSE、TSP2HD和Sj14-3-3。IgM的敏感性高于IgG。免疫印迹中，感染后1周小鼠血清可识别SEA约73 kDa的蛋白条带，且随着感染时间延长反应增强。此外，感染后第10天小鼠血清最早识别GST-HD的条带（33 kDa），该条带与感染后5周小鼠血清反应强烈。
GST-HD融合蛋白及SEA中分子量约73 kDa的蛋白对血吸虫病具有早期诊断价值，免疫印迹的敏感性高于ELISA。
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