中国寄生虫学与寄生虫病杂志 ›› 1995, Vol. 13 ›› Issue (2): 142-145.

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阴道毛滴虫整虫抗原和单克隆抗体进行斑点酶联免疫吸附试验条件的探讨

于超,高兴政,朱永红   

  1. 北京医科大学寄生虫学教研室
  • 出版日期:1995-05-31 发布日期:2017-01-05

EXPLORATION ON EXPERIMENTAL CONDITIONS OF DOT ELISA FOR TRICHOMONAS VAGINALIS USING INTACT PARASITE ANTIGEN MONOCLONAL ANTIBODY DETECTING SYSTEM

  • Online:1995-05-31 Published:2017-01-05

摘要: 以阴道毛滴虫整虫抗原和单克隆抗体进行dot-ELISA试验,分别观察了不同的虫体数量、孵育时间、封闭剂型、抗原类型、NC膜及抗原保存时间与温度等条件对试验效果的影响。结果显示以2.5×107细胞/ml的虫数制备抗原膜,用0.5%脱脂奶粉1次性封闭1h(37℃),单克隆抗体和酶分别孵育1h(37℃)为最佳条件。在4℃和-20℃温度下,抗原可保存4个月。

关键词: 阴道毛滴虫, 斑点酶联免疫吸附试验, 整虫抗原

Abstract: This paper deals with the effect of different experimental conditions including the number of parasite, blocking fluid, incubation time and different types of antigens on dot ELISA for Trichomonas vaginalis. Five microlitres of freshly washed intact worms at a concentration of 7.7×10 5-3×10 8 cells/ml were added onto nitrocellulose membrane blocked with 1% BSA, 10% bovine serum or milk power (0.1%,0.25%,0.5%,5%). Dot ELISA using nitrocellulose membrane containing whole cell antigen 2.5×10 7 cells/ml, blocked by 0.5% defatted milk power at 37℃ for one hour before the incubation in McAb, incubated at 37℃ for one hour in McAb and SPA respectively was found to give more promising results. The whole cell antigen could be preserved for four months at 4℃ and -20℃ .

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