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当前位置: 首页 > 文章 > Dot-ELISA检测H9亚型禽流感病毒的研究 中国畜牧兽医 2008,35 (10) 87-89
Position: Home > Articles > Study on Detection of Avian Influenza Virus H9 Subtype by Dot-ELISA China Animal Husbandry & Veterinary Medicine 2008,35 (10) 87-89

Dot-ELISA检测H9亚型禽流感病毒的研究

作  者：
焦显芹;廖仲磊;陈红英;韩玉林;王磊;李新生
单  位：
河南省荥阳市畜牧局;河南省郑州市现代农业科技服务中心;河南省焦作市畜牧局;河南农业大学牧医工程学院
关键词：
Dot-ELISA;检测;H9亚型;禽流感病毒
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。
译  名：
Study on Detection of Avian Influenza Virus H9 Subtype by Dot-ELISA
作  者：
JIAO Xian-qin1,LIAO Zhong-lei2,CHEN Hong-ying2,3,HAN Yu-lin4,WANG Lei5,LI Xin-sheng2,3(1.Zhengzhou Modern Agricultural Technology Service Centre,Zhengzhou 450002,China;2.The College of animal Husbandry And Veterinary,Henan Agricultural University,Zhengzhou 450002,China;3.Animal Food Safety Key Laboratory,Henan Province,Zhengzhou 450002,China;4.Husbandry and Veterinary Burearu of Jiaozuo,Jiaozuo 454150,China;5.Husbandry and Veterinary Burearu of Xingyang,Xingyang 450100,China)
关键词：
Dot-ELISA;detection;H9 subtype;avian influenza virus
摘  要：
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 economical,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.

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