【摘要】 目的以鸡卵黄免疫球蛋白IgY代替哺乳动物来源的IgG抗体,建立ELISA双抗体夹心法,探索用于诺如病毒检测的可行性。方法以387型菌株(GⅡ.4型)诺如病毒P颗粒为抗原免疫健康产蛋来航鸡,制备、纯化特异性抗诺如病毒鸡卵黄免疫球蛋白IgY。采用过碘酸钠法对IgY进行酶标。以特异性抗体为捕获抗体,酶标抗体为检测抗体,建立双抗体夹心ELISA检测体系。评价其灵敏度、特异度及重复性。并将其与金标准进行比较。结果本实验建立的双抗体夹心ELISA检测体系最低检测限达20 ng/ml,批内变异系数为1.021%,批间变异系数为1.150%。临床应用评价显示真阳性率为82.5%,真阴性率为81.82%。与金标准比较,检出率差异无统计学意义(P=0.629)。结论建立了基于特异性抗诺如病毒卵黄抗体IgY的双抗体夹心ELISA检测体系,用于腹泻患者粪便标本诺如病毒的检测具有较好的敏感性和特异性。更多还原

【Abstract】 Objective To establish the enzyme-linked immunosorbent assay(ELISA) double antibodies sandwich method by using chicken egg yolk immunoglobulin(IgY) instead of mammalian IgG antibodies for the detection of norovirus.Methods Nov-specific IgY were produced by immunizing healthy laying Leghorn chickens with type 387 Nov P particles(Type GⅡ.4).IgY was labeled with HRP by sodium iodide method.The ELISA double antibodies sandwich method was established with the specific antibodies as capture antibodies and the enzyme-antibodies as the detection antibodies.The sensitivity,specificity and reproducibility were evaluated and also compared with the gold standard for clinical application evaluation.Results The ELISA double antibodies sandwich method was established.The minimum detection level was 20 ng / ml,the intra-assay coefficient of variation was 1.021% and the inter-assay coefficient was 1.150%.The evaluation of clinical application showed the true positive rate was 82.5%,and the true negative rate was 81.82%.Compared with the gold standard,the detection rate has not significant difference(P>0.05).Conclusions The ELISA double antibodies sandwich method based on Nov-specific egg yolk antibodies(IgY) was established successfully.It has high sensitivity and specificity for the detection of diarrhea stool specimens.更多还原