Dot-ELISA检测H9亚型禽流感病毒的研究  

Study on Detection of Avian Influenza Virus H9 Subtype by Dot-ELISA

作　　者：焦显芹 廖仲磊[2] 陈红英[2] 韩玉林 王磊[4] 李新生[2] 

机构地区：[1]河南省郑州市现代农业科技服务中心,郑州450002 [2]河南农业大学牧医工程学院,郑州450002 [3]河南省焦作市畜牧局,焦作454150 [4]河南省荥阳市畜牧局,荥阳450100

出　　处：《中国畜牧兽医》2008年第10期86-88,共3页China Animal Husbandry & Veterinary Medicine

基　　金：国家十一五科技支撑计划(2006BAD06A08)

摘　　要：本研究采用兔抗H9亚型禽流感病毒(AIV)IgG包被于硝酸纤维素膜,酶标羊抗兔IgG作二抗,建立检测H9亚型AIV的Dot-ELISA法。经方阵试验确定兔抗AIVIgG工作浓度为1∶400,酶标羊抗兔IgG的工作浓度为1∶400。作者建立的Dot-ELISA对AIV的最小检测量为3.35×10-9g。Dot-ELISA与HA和HI、AGP及病毒分离法相比,检测63份临床疑似H9亚型AIV病料,Dot-ELISA检出32份(57.14%),HA和HI检出15份(23.81%),AGP检出11份(17.46%),病毒分离检出38份(60.30%)。用抗H9亚型AIV阳性血清可以阻断Dot-ELISA阳性反应,诊断膜片与鸡新城疫病毒、鸡传染性法氏囊病病毒、鸡传染性支气管炎病毒、产蛋下降综合征病毒不出现阳性反应,证明Dot-ELISA特异性好。分别置室温(25℃左右)、4℃和-20℃下保存1个月后,膜片诊断效果不变,对照反应均成立,该方法重复性好(重复符合率为93.9%),操作简便(3 h内可完成),不需要特殊检测仪器,结果客观,肉眼易于判断,是微生物和传染病及寄生虫病诊断标准化的新技术之一。A Dot-ELISA method was developed for the detection of avian influenza virus （AIV） H9 subtype. The optimum working concentration of rabbit anti-AIV IgG was determined to be 1 ： 400,and that of goat anti-rabbit IgG labeled with horseradish peroxidase（HRP）to be 1 ： 400 too. The assay could detect as low as 3.35× 10^-9 g/disc. Comparison of the Dot-ELISA with HA and HI, AGP, virus isolation were conducted. In the Dot-ELISA, 36 of 63 were positive （57.14%）, 38 of 63 in the virus isolation were positive （60. 30%）. The high specify of the Dot-ELISA was shown by the specific blocking test with AIV positive serum and cross-reaction test with newcastle disease virus, infectious bronchitis virus, infectious bursal disease virus, egg drop syndrome virus. The result of test reproducibility in the Dot-ELISA was very good （93.9%）. The procedure took about 3 h, and it was gconomical, results of reaction could be easily read by eye. This rapid and inexpensive method could be proved to be a new diagnostic technique for early diagnosis of avian influenza.

关 键 词：DOT-ELISA 检测 H9亚型 禽流感病毒 

分 类 号：S852.659.2[农业科学—基础兽医学]