鸽毛滴虫感染Dot-ELISA检测方法的建立  

Development of Dot-ELISA for detecting Trichomonas gallinae from pigeon

作　　者：罗锋[1,2,3] 郭炳新[1] 李国清[1] 苏遂琴[4] 梁刚[4] 陈泽华[4] 

机构地区：[1]华南农业大学兽医学院,广东广州510642 [2]华南农业大学动物科学学院,广东广州510642 [3]广东温氏食品集团研究院,广东新兴527400 [4]佛山出入境检验检疫局,广东佛山528200

出　　处：《中国兽医杂志》2010年第10期18-20,共3页Chinese Journal of Veterinary Medicine

基　　金：华南农业大学校长基金项目(2008B006);广东出入境检验检疫局科技计划项目(2006GDK03);教育部博士点基金项目(200805640004)

摘　　要：为了制备兔抗鸽IgG抗体-HRP,建立斑点酶联免疫吸附试验定性检测鸽毛滴虫抗原的检测方法,将纯化的鸽IgG加入佐剂免疫兔子,获取纯化的兔抗鸽IgG抗体;采用简易过碘酸钠法标记兔抗鸽IgG,获取兔抗鸽IgG抗体-HRP;同时将鸽毛滴虫用超声波粉碎加入佐剂后多次免疫健康鸽获取鸽抗鸽毛滴虫高免血清(一抗);然后在2×106个/mL的鸽毛滴虫抗原量下,用不同浓度的一抗抗体、兔抗鸽IgG抗体-HRP进行试验;并对70个临床样品进行检测。结果显示,在2×106个/mL的鸽毛滴虫抗原量下,一抗与兔抗鸽IgG-HRP最佳工作浓度分别为1∶100和1∶500;70个样品中,48个镜检为阳性的样品用Dot-ELISA检测有44个呈阳性,阳性符合率为91.7%;22个镜检为阴性的样品用Dot-ELISA检测有12个呈阴性,阴性符合率为54.5%;Dot-ELISA检测与镜检结果的总符合率为80%。结果表明,本试验成功地建立了鸽毛滴虫感染的Dot-ELISA检测方法。To obtain the rabbit anti-pigeon IgG-HRP and develop a Dot-ELISA method for detecting T.gallinae from pigeon,firstly,the rabbits were injected with the purified pigeon IgG in adjuvant,and the purified rabbit anti-pigeon IgG antibody were obtained.Then the rabbit anti-pigeon IgG-HRP were prepared after labeled with facility prosodium iodate.At the same time,some pigeons were injected with the parasite vaccine in adjuvant,then the pigeon anti-T.gallinae serum were obtained.At the parasite number 2×10^6/mL,Dot-ELISA test was conducted using the different concentrations of pigeon anti-T.gallinae serum and anti-pigeon-HRP conjugate,and 70 clinic samples from pigeons were detected.The results showed that when the parasite number was 2×10^6/mL,the optimal working concentrations of anti-T.gallinae antiserum and IgG-HRP were 1∶100 and 1∶500,respectively,44 positive and 12 negative in total 70 samples were detected with Dot-ELISA, while 48 positive and 22 negative samples were detected with wet film examination under microscope.The positive rate,the negative rate and the total conformity rate were 91.7%,54.5% and 80%,respectively.The results indicated that the Dot-ELISA method for detecting T.gallinae antigen from pigeon was developed successfully.

关 键 词：鸽毛滴虫 兔抗鸽IgG抗体-HRP 鸽抗鸽毛滴虫高免血清 DOT-ELISA 

分 类 号：S852.72[农业科学—基础兽医学]