纳米金层析免疫法快速检测黄热病病毒

目的:探索快速、简便、高效、灵敏、低廉地检测黄热病病毒感染的方法。方法:用纳米金免疫层析试验原理研制了一种快速诊断试纸。将制备的纳米金颗粒用SPA标记形成金-SPA复合物,其复合物结合在聚酯膜上制成金标结合释放垫;通过基因工程技术在大肠杆菌中克隆和表达黄热病E蛋白,经纯化的抗原蛋白被固定在硝酸纤维素膜上,然后滴加待测血清,样品中的病毒抗体通过免疫反应与膜上的抗原蛋白结合,待其渗过膜片后,与金-SPA复合物结合形成肉眼可见的红色线条。结果:用已确证的21份黄热病阳性血清和30份阴性血清进行了试验,该快速检测方法与ELISA方法无显著差异。结论:该检测方法不需任何仪器,仅凭肉眼即可判定结果,整个检测过程不超过5min。与传统的ELISA法相比,具有方便快速、成本低廉、应用范围广等优点。

Rapid and Simultaneous Detection of Yellow Fever Virus by Nano-Gold Immunochromatography Assay

Objective:To design a colloidal gold method for rapid detection of yellow fever(YF),which is fast,simple,efficient,sensitive,and inexpensive.Methods:Based on colloidal gold immunochromatography assay(GICA),we established a diagnostic test strip for the rapid detection.The colloidal gold particles were labeled by streptavidin(SPA)to form gold-SPA complex.Then the gold-SPA complex was combined onto a polyester film.The E proteins of YF viruses were cloned and expressed in E.coli using genetic engineering techniques.The purified proteins were fixed on a nitrocellulose membrane.When the serum sample was dropped onto the membrane,the virus antibody will bind with the antigen on the membrane by immune response.After permeation through the membrane,the antibody-antigen complex will bind with colloidal gold-SPA to form a red line visible to the naked eye.Results:This method was tested by 21confirmed YF positive serum samples and 30negative serum samples.The results showed that there was no significant difference between the rapid detection method and ELISA method.Conclusion:The rapid detection method does not require any instrument and the result can be determined by naked eye in less than 5minutes.Compared with ELISA,this rapid detection method is convenient,fast,inexpensive,and widely applicable.