检测蓝氏贾第鞭毛虫病毒表达的绿色荧光蛋白间接ELISA方法的建立  

ESTABLISMENT OF INDIRECT ELISA FOR DETECTION OF GREEN FLUORESCENT PROTEIN EXPRESSED BY GIARDIAVIRUS

作　　者：刘全[1] 张西臣[1] 李建华[1] 尹继刚[1] 赵永军[1] 

机构地区：[1]解放军军需大学军事兽医系,吉林长春130062

出　　处：《中国寄生虫病防治杂志》2005年第2期85-87,共3页Chinese Journal of Parasitic Disease Control

基　　金：国家自然科学基金资助项目(No.30300260)。

摘　　要：目的建立蓝氏贾第鞭毛虫病毒(GLV)介导的绿色荧光蛋白(GFP)表达定量检测方法,为GFP在寄生性原虫病毒研究中的应用奠定基础。方法以GFP兔抗血清为一抗,辣根过氧化物酶标记的羊抗兔血清为二抗,建立GFP定量检测的间接ELISA方法。结果定量检测GFP的间接ELISA方法最适反应条件为以10%正常胎牛血清作为封闭剂,一抗的最佳稀释度为1∶3200,二抗的最佳稀释度为1∶1600。结论以GFP兔抗血清为一抗成功建立GFP定量检测的间接ELISA方法,检测结果能正确反映GLV介导的GFP表达情况。Objective To establish indirect ELISA for quantitative detection of green fluorescent protein(GFP) expressed by Giardiavirus. Methods Rabbit anti-GFP serum was used as the first antibody and goat anti-rabbit IgG-HRP was used as the second antibody, the best dilution for rabbit anti-GFP serum and goat anti-rabbit IgG-HRP and the blocking agent were decided according to the best reaction condition standards for ELISA, the indirect ELISA for detection of GFP expressed by Giardiavirus was established. Results Blocking agent was 10% normal foetus cattle serum, the best dilution of first and second antibody were 1∶3 200 and 1∶1 600 respectively. Conclusion The indirect ELISA in which rabbit anti-GFP serum was the first antibody precisely reflected GFP expressed by GLV in Giardia lamblia.

关 键 词：贾第虫病毒 绿色荧光蛋白 ELISA 

分 类 号：R382.2[医药卫生—医学寄生虫学]