胶体金免疫层析法检测猪瘟(强弱毒区分)抗体研究

以原核表达的重组猪瘟病毒E2蛋白为弱毒抗原,以猪瘟病毒(HCV)强毒株细胞培养物为强毒抗原,利用胶体金标记技术国内首次建立了HCV抗体检测(强弱毒区分)的免疫层析试纸条(双抗原夹心法)。研究结果表明,检测时只需将80μL待检测猪血清加于加样端20 min后观察结果即可。若观察区上端仅1条红色条带,即为HCV抗体阴性;2~3条条带即为HCV抗体阳性;若出现2条条带的位置为观察区上端和中间部位,即为HCV弱毒抗体阳性;若出现2条条带的位置位于观察区上端和下端部位,则为HCV强毒抗体阳性;若出现3条条带则HCV强弱毒抗体均为阳性。本试纸可很好地避免因试验操作造成的假阳性、假阴性,且操作简单、快速、简便、灵敏度高、特异好、不需特殊设备、价格低廉,20 min即可判断结果,且可区分强弱毒感染,适于基层实验室和现场检测。

Study on gold immunochromatography assay for detection of hog cholera antibody

Recombination Hog cholera virus E2 protein which was expressed by prokaryotic was used as attenuated antigen,Hog cholera virus(HCV)was served as virulent HCV antigen.HCV antibody test of immunochromatographic test strip was established for the first time in China(double sandwich method).80 μL porcine serum was added to the end of strips.One red band meant HCV antibody was negative,two or three bands meant HCV antibody was positive.It was HCV attenuated antibody positive if two bands were on the top and middle of the observation area;It was HCV virulent antibody positive if two bands were on the top and bottom of the observation area;If there were three bands,the attenuated and strong antibody were both positive.The method could discriminate between virulent and attenuated strain,which was more sensitive and specific than other methods and fit for clinical diagnosis of HCV.