摘要
为建立一种简便、快速、特异的H7亚型禽流感病毒抗体检测方法,将荧光微球偶联标记鼠抗鸡IgG,并将羊抗鼠IgG和重组H7 亚型禽流感病毒HA蛋白分别作为质控线(C线)和检测线(T线),喷涂于硝酸纤维素膜上.采用单一变量分析法对反应体系进行了优化,随后对该方法的敏感性、特异性、重复性及符合率进行了评估.结果显示:本研究建立的H7 亚型禽流感病毒荧光微球抗体检测方法比参照国标建立的血凝抑制试验灵敏度高 2 倍;特异性较好,对H5 亚型禽流感病毒、H9 亚型禽流感病毒、鸡新城疫病毒、鸡传染性支气管炎病毒、鸡传染性法氏囊病病毒和鸡减蛋综合征病毒等常见禽病病原阳性血清的检测结果均为阴性;重复性试验结果的变异系数为 3.13%～8.46%,具有较好的重复性;本研究方法和血凝抑制试验血清抗体检测结果的总体符合率为 95.67%,两种方法具有较高的一致性.结果表明,本研究建立的H7 亚型禽流感病毒荧光微球抗体检测方法性能稳定、结果可靠,为H7 亚型禽流感快速检测提供了新方法.
Abstract
In order to establish a simple,rapid and specific method to detect antibodies against H7 subtype avian influenza virus(AIV),mouse anti-chicken IgG was coupled with fluorescent microspheres,and sheep anti-mouse IgG and HA protein of recombinant H7 subtype AIV were sprayed on nitrocellulose membrane as quality control line(C line)and detection line(T line),respectively.The reaction system was optimized by single variable analysis,and its sensitivity,specificity,reproducibility as well as coincidence rate with hemagglutination inhibition(HI)test were evaluated.The results showed that the established method was twice more sensitive than the HI test established with reference to national standard;good specificity was observed,as it can't react with H5 subtype AIV,H9 subtype AIV,Newcastle disease virus(NDV),infectious bronchitis virus(IBV),infectious bursal disease virus(IBDV),egg drop syndrome virus(EDSV)and other common positive serum of avian disease pathogens;the variable coefficients of reproducibility test ranged from 3.13%to 8.46%,indicating a good repeatability;and the results detection by the established method were 95.67%consistent with those by HI test,indicating a high consistency.In conclusion,the established method with stable performance and reliable results could be used to rapidly detect antibodies against H7 subtype AIV as a new tool.
维普
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