摘要
对禽流感病毒H9亚型血凝素单克隆抗体4C4进行胶体金标记,喷涂于玻璃纤维上制成金标垫,鸡抗AIVIgG和羊抗鼠IgG分别包被于硝酸纤维素(NC)膜上作为检测线和质控线,制作禽流感H9亚型免疫层析检测试纸条。验证结果表明,试纸条操作简单,肉眼于10 min内可判定结果,对H9亚型禽流感病毒及其标准血凝抗原具有高度特异性,与其它亚型禽流感病毒标准抗原及其它禽类病毒没有交叉反应,检测灵敏度比HA试验高4倍以上。试纸条在室温保存6个月、4℃保存12个月,其特异性及灵敏度没有明显变化。对实验室送检及临床采集的共311份喉头拭子进行检测,结果与本室研制的禽流感ELISA试剂盒总符合率为92%,随机选取10份阳性样品经病毒分离证实无一误检。结果证明,本研究建立的H9亚型禽流感病毒免疫层析检测方法具有特异、敏感、稳定、操作简单快捷等特点,适合用于H9亚型禽流感病毒的现场检测及鉴别诊断。
An immunochromatographic strip was developed for the detection of the H9 subtype of avian influenza viruses （H9AIVs） in poultry using McAb 4C4 for H9AIV hemagglutinin and chicken anti-AIV antibody. The 4C4 was labeled with colloidal gold as a detection reagent, and chicken anti-AIV antibody was blotted on the test line while a goat anti-mouse antibody was used on the control line of the nitrocellulose membrane. In comparison with the hemagglutination （HA） and hemagglutination inhibition （HI） test, the strip was specific for detecting HgAIV with the sensitivity at 0. 25 HA units within 10 min. Storage of the strips at room temperature for six months or at 4 ℃ for 12 months had not changed their sensitivity and specificity. 311 tracheal samples from potentially infected chickens were detected using the strip and ELISA kit established in our lab, the coincidence ratio was 92%. Further characterization of 10 positive samples randomly selected showed that no single sample was false positive, as determined by the HI assays and standard virus isolation. Our research revealed that the immunochromatographic strip for detection of H9AIVs has high specificity, sensitivity and stability. This, together with the advantages of rapid detection and easy operation without requirement for special skills and equip- ments, makes it suitable for the on-site detection and differentiation of H9AIVs from other viruses in poultry.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2008年第8期1081-1086,共6页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
湖北省科技攻关重大专项(2006AA202A05)
湖北省科技攻关禽流感应急专项(2005AA401D60)
