鸡传染性法氏囊病双抗体夹心Dot─ELISA诊断法的建立及应用  

Establishment and Application of Sandwich Dot-ELISAfot the Detection of Infectious Bursal Disease Virus

作　　者：张春杰[1] 郑祥海[1] 程相朝[1] 

机构地区：[1]洛阳农业高等专科学校

出　　处：《中国兽医学报》1994年第2期164-167,共4页Chinese Journal of Veterinary Science

摘　　要：以醋酸纤维素膜作为固相载体，辣根过氧化物酶标记鸡抗传染性法氏囊病病毒抗体（ＩＢＤ—ＩｇＧ），饱和二氨基联苯胺为底物显色，建立了鸡传染性法氏囊病双抗体夹心Ｄｏｔ—ＥＬＩＳＡ诊断法。经方阵实验确定最佳反应条件为：ＩｇＧ的包被液为０．０５ｍｏｌ／Ｌ（ｐＨ９．６）碳酸盐缓冲液，包被浓度为１：５０；酶标抗体的工作浓度为１：１００；洗涤液为含０．０５％吐温—８０的０．０２ｍｏｌ／Ｌ（ｐＨ７．４）磷酸盐缓冲液；封闭液为含０．２％明胶的洗涤液；封闭时间、抗原及酶标抗体的作用时间均为３７℃３０ｍｉｎ。应用本方法和琼扩试验同步检测２０份已知阳性病料、１２０份待检病料和胚毒尿囊液、１０份正常鸡样品，结果表明，Ｄｏｔ—ＥＬＩＳＡ阳性率为９０％，而琼扩试验为４０％；凡琼扩试验阳性者，Ｄｏｔ—ＥＬＩＳＡ均呈强阳性，而在Ｄｏｔ—ＥＬＩＳＡ阳性样品中，只有４４％呈琼扩试验阳性，Ｄｏｔ—ＥＬＩＳＡ的敏感度为琼扩试验的１００倍。The Sandwich Dot-ELISA,a rapid and sensitive method,was estab-lished to detect antigen of infectious bursal disease virus, 150 samples were syn-chronously tested by Dot-ELISA and AGP,The results Showed that the Dositive ratesof Dot-ELISA and AGP were 90%and 40%respectively. All the positive samples de-tected by AGP were also positive by Dot-ELISA, whereas among the positive samplesby Dot-ELISA,only 44%of them revealed positive reaction in AGP,The sensitivity ofDot-ELISA was 100-fold as high as AGP.

关 键 词：鸡 DOT-ELISA 抗原检测 IBD 

分 类 号：S858.315.3[农业科学—临床兽医学]