O型口蹄疫病毒免疫层析试纸条检测方法的建立

Development of a Rapid Chromatographic Strip for the Diagnosis of Serotype O Foot-and-mouth Disease Virus

为建立一种快速、准确检测O型口蹄疫病毒(FMDV)抗原的胶体金免疫层析方法,将兔、豚鼠抗O型FM-DV多抗用DEAE-Sephose层析柱纯化。胶体金标记O型豚鼠口蹄疫抗体,形成金标探针并将其喷涂于玻璃纤维上。兔抗O型FMDV抗体和羊抗豚鼠IgG分别标记于硝酸纤维素膜上作为检测带和质控带,各部件按顺序装配形成快速诊断试纸条。如果待检样品中含有O型FMDV,它将与玻璃纤维上的胶体金探针和兔抗O型FMDV抗体形成夹心复合物,并在检测带被固定,沉集反应形成肉眼可见的红色条带。在田间试验中,53份试验样本分别用试纸条和反向间接血凝试验进行检测,2种方法的阳性率分别为95.45%和90.91%。评价试验证实,本研究建立的胶体金免疫层析方法简便、快速,具有良好的特异性和敏感性,非常适于基层兽医实验室诊断时使用。

To develope a rapid and accurate test for detecting serotype O foot-and-mouth disease virus（FMDV）based on the principle of gold immunochromatographic assay（GICA）,guinea pig and rabbit polyclonal antibody against FMDV serotype O were purified by DEAE-sephrose.The guinea pig anti（FMDV） serotype O-IgG was conjugated with colloidal gold,which was called gold probes,and was dispensed on the glass fiber.The rabbit anti（FMDV） serotype O-IgG and the goat anti-guinea pig IgG polyclonal antibody were separately sprayed on the nitrocellulose membrane as the test line（T line） and the control line（C line）.The rapid gold immunochromatographic strip was assembled by the different accessory in regular sequence.If there was serotype O FMDV in sample,gold probes on the glass fiber and rabbit anti（FMDV） serotype O-IgG formed a sandwich complex with serotype O FMDV.Within the test line,the sandwich complex was immobilized, furthermore,concentrated and appeared a distinct red color on the test line.In the clinic test,a total of 53 field samples were comparatively detected with both GICA and reverse indirect hemagglutination assay（RIHA）,and the positive rate were 95.45% and 90.91%,respectively.The evaluation test proved GICA established in this study is simple,rapid,sensitive and specific.It is suitable for basic veterinary medical laboratories.