预警哨鼠抗Sj23HD IgG抗体Western blot检测方法的改进及应用

发布时间：2018-06-26 05:30

本文选题：血吸虫 + 日本　；参考：《中国病原生物学杂志》2013年12期

【摘要】：目的改进检测预警哨鼠血清抗日本血吸虫23ku膜蛋白大亲水肽段(Sj23HD)IgG抗体的Western blot方法,并观察其现场应用价值。方法预先制备转印有重组Sj23HD蛋白的NC膜条,采用化学发光Western blot检测哨鼠血清抗Sj23HD IgG,观察方法的稳定性和敏感性,并对2012年江苏省潜在血吸虫感染高风险水域监测预警哨鼠回收后第14、21、35d血样进行检测,同时以ELISA检测方法作为对照,对免疫检测结果与解剖查虫结果进行统计学分析。结果改进后的Western blot操作简便,比常规方法节省1~2周时间。预制NC膜条至少可低温保存6个月,其敏感性与新鲜制备的NC膜条的敏感性相似。使用改进后的Western blot方法检测预警哨鼠14、21和35d血清标本,阳性率分别为0.90%,3.50%和5.06%,ELISA阳性率分别为0.45%、3.00%和5.06%,差异无统计学意义(P0.05)。两种免疫学方法的敏感性均高于解剖查虫法。结论改进后的Western blot操作简便,敏感性高,可用于预警哨鼠血吸虫感染检测。
[Abstract]:Objective to improve the Western blot method for detection of anti-Schistosoma japonicum 23ku membrane protein large hydrophilic peptide fragment (Sj23HD) IgG antibody in early warning sentinel rat serum, and to observe its application value in the field. Methods the NC membrane strips with recombinant Sj23HD protein were prepared in advance. The serum anti-Sj23HD IgG of sentinel mice was detected by chemiluminescence Western blot. The stability and sensitivity of the method were observed. The blood samples of sentinel rats collected from potential Schistosoma japonicum infection high risk areas in Jiangsu Province in 2012 were detected on day 1421, and Elisa was used as control. The results of immunoassay and dissection were analyzed statistically. Results the improved Western blot was easy to operate and saved 1 ~ 2 weeks compared with the conventional method. The prefabricated NC strips can be preserved at least 6 months at low temperature, and the sensitivity of prefabricated NC strips is similar to that of freshly prepared NC strips. The positive rates of Elisa were 0.90% and 5.06%, respectively, and the positive rates were 0.45% and 5.06%, respectively. There was no significant difference between the two groups (P0.05). The sensitivity of the two immunological methods was higher than that of the dissecting method. Conclusion the improved Western blot is simple and sensitive, and can be used to detect schistosomiasis infection in sentinel mice.
【作者单位】：卫生部重要寄生虫病预防与控制技术重点实验室江苏省寄生虫分子生物学重点实验室江苏省血吸虫病防治研究所;卫生部重要寄生虫病预防与控制技术重点实验室江苏省寄生虫分子生物学重点实验室江苏省血吸虫病防治研究所;卫生部重要寄生虫病预防与控制技术重点实验室江苏省寄生虫分子生物学重点实验室江苏省血吸虫病防治研究所;
【基金】：国家传染病重大专项(No.2012ZX10004-220) 国家自然科学基金项目(No.30972581,81201316) 江苏省自然科学基金项目(No.BK2008110,BK2012544) 江苏省卫生厅科技项目(No.H200738,H201066) 江苏省医学重点人才项目(No.RC2011094) 江苏省兴卫工程基金项目(No.ZX201108) 江苏省血吸虫病、地方病及寄生虫病项目(No.X201115)
【分类号】：R392.1

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