摘要
用微波法制备金溶胶,对禽流感病毒(AIV)H3和H7亚型单克隆抗体(H3-1D6和H7-1F7)用亲和层析法进行纯化,优化单抗与金溶胶的最佳结合条件后,喷涂于玻璃纤维上制成金标垫。将纯化的马抗H3和H7亚型AIV多克隆抗体和山羊抗鼠二抗分别包被于硝酸纤维素(NC)膜上作为检测线和质控线,制备胶体金检测试纸条。用制备的试纸条对H3和H7亚型AIV标准抗原、毕赤酵母真核表达蛋白和已知样品进行检测,结果与血凝试验、血凝抑制试验、AC-ELISA和RT-PCR方法相符。同时用该方法对H5和H9亚型AIV标准抗原、病料以及传染性支气管炎、新城疫等抗原进行检测,结果均为阴性。该方法操作简单,肉眼于10min内可判定结果,且达到了血凝试验和血凝抑制试验的敏感性。
Au-collosol was prepared by microwave, purified the monoelonal antibody of H3 and H7 subtypes AIV （H3-1D6 and H7-1F7）, and optimized the combine condition, then, sprayed on the fiberglass, and obtained Aumarked pad. Coated the purified horse-anti-H3 and H7 polyclonal antibody on nitrocellulose membrane as the detec- tion line,and goat-anti-Mus antibody as the quality control line. Detected the standard antigen of H3 and H7, ex- pressed protein in Pichia pastoris, and the known sample, the result showed conformity with the hemagglutination test, hemagglutination inhibition assay, AC-ELISA and RT-PCR. Detected the standard antigen of H5 and H9,nega- tive samples, IBV, NDV simultaneously, the result showed negative. This method has a simple operation, which could assess the result within 10 minutes,and the sensitivity was the same as hemagglutination test and hemagglutination inhibition assay.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2009年第5期580-583,共4页
Chinese Journal of Veterinary Science
基金
科技部“十五”攻关资助项目(2004BA519A15)
关键词
禽流感病毒
H3亚型
H7亚型
胶体金
avian influenza viruses
H3 subtype
H7 subtype
gold immunochromatography
