吉林大学学报(理学版)

• 生命科学 • 上一篇    

轮状病毒间接免疫荧光的检测方法

姜涛, 吴丛梅, 刘新涛, 金浩, 殷玉和   

  1. 长春工业大学 化学与生命科学学院, 长春 130012
  • 收稿日期:2015-11-11 出版日期:2016-07-26 发布日期:2016-07-20
  • 通讯作者: 殷玉和 E-mail:yyhlxt72@163.com

Detection Method of Indirect Immunofluorescence Assay for Rotavirus

JIANG Tao, WU Congmei, LIU Xintao, JIN Hao, YIN Yuhe   

  1. School of Chemistry and Life Science, Changchun University of Technology, Changchun 130012, China
  • Received:2015-11-11 Online:2016-07-26 Published:2016-07-20
  • Contact: YIN Yuhe E-mail:yyhlxt72@163.com

摘要:

以兔抗SA11多克隆抗体为一抗、 Alexa488标记的山羊抗兔抗体为二抗, 通过对反应条件的优化建立轮状病毒(RV)检测的间接免疫荧光法(IFA). 实验结果表明, IFA最佳工作条件为: MA104细胞密度为每孔2×104个, 胰酶活化轮状病毒质量浓度为1 μg/mL, 培养时间为18 h, 一抗最适稀释度为1∶800, 二抗最适稀释度为1∶500.

关键词: 轮状病毒, 间接免疫荧光, 胰酶活化, 滴度

Abstract:

Using rabbit polyclonal antibody against SA11 as the primary antibody and Alexa488 labeled the goat antirabbit antibody as the secondary antibody, we established an indirect immunofluorescence assay (IFA) for detection of Rotavirus (RV) through the optimization of reaction conditions. Experimental results show that the optimum conditions of IFA are as follows: the best density of MA104 cells is 2×104 cells per well, the best mass concentration of trypsin activation rotavirus is 1 μg/mL, the best time of incubate Rotavirus is 18 h, and the optimum working dilutions of the rabbit polyclonal antibody against SA11 and Alexa488 labeled the goat antirabbit antibody are 1∶800 and 1∶500, respectively.

Key words: Rotavirus; indirect immunofluorescence assay, trypsin activation, titer

中图分类号: 

  • R392