猪瘟病毒胶体金免疫层析检测方法的建立及应用

为建立一种猪瘟病毒的快速检测方法,采用柠檬酸三钠还原氯金酸制备胶体金,选择15 nm胶体金标记兔抗猪瘟病毒抗体,对胶体金标记抗体采用低温超速离心法进行纯化,组装免疫层析试纸条。用试纸条对猪瘟可疑病料进行检测,同时用直接荧光抗体试验和RT-PCR做平行试验。结果显示,用试纸条检测猪瘟病毒,10 min左右即可显示结果,试纸条、直接猪瘟荧光抗体试验和RT-PCR的阳性检出率依次为36.36%(4/11)、45.45%(5/11)和72.72%(8/11)。表明,用胶体金免疫层析试纸条检测猪瘟病毒操作简便,需时短,可以用于猪瘟的流行病学调查。

Establishment and application of colloidal gold-based immunochro matographic assay for the detection of classic swine fever virus

To establish a rapid method for the detection of classic swine fever virus(CSFV),tri-sodium citrate with aqueous gold chloride were warmed up and mixed to synthesize gold sol.The 15 nm colloidal gold particles were selected from colloidal gold nanoparticles.The polyclonal antibody against CSFV was labeled with the selected gold colloid.To assemble an immunochromatographic strip,the labeled rabbit IgG against CSFV was purified by ultra-centrifugation at low temperature. The clinical samples of CSFV were examined by the assembled immunochromatographic strip,while they were also examined by both the fluorescent antibody test and RT-PCR.It taken only 10 min to finish the examination of the clinical CSFV samples by the immunochromatographic strip,and the positive rates of the clinical CSFV samples which were detected by the immunochromatographic strip,the fluorescent antibody test and RT-PCR were 36.36%(4/11),45.45%(5/11) and 72.72%(811),respectively.The results showed that the detection of CSFV was easily and quickly carried out by the immunochromatogrqphic strip assembled with the selected gold colloid,and the strip can be applied to the epidemiologic investigation of CSFV.