基于纳米磁珠免疫层析技术建立登革病毒快速检测方法

Establishment of rapid detection method for dengue virus based on nano-magnetic beads immunochromatography

目的基于免疫层析技术,利用纳米磁珠建立登革病毒抗原快速检测方法。方法用登革病毒检测抗体包被纳米磁珠制作免疫探针,在硝酸纤维膜上喷涂登革病毒捕获抗体做检测线,喷涂羊抗鼠多克隆抗体做质控线。将重组登革病毒NS1蛋白稀释至100、50、20、10、1、0.1 ng/ml,用建立的纳米磁珠免疫层析试纸条进行测试,同时用市售胶体金试纸条进行对比。结果通过肉眼观察,纳米磁珠免疫层析试纸条检测登革病毒NS1蛋白的检测限是1 ng/ml,市售胶体金试纸条检测限是10 ng/ml。经特异性检测,和西尼罗病毒、寨卡病毒假病毒、埃博拉病毒、甲型流感病毒没有交叉反应。结论建立了登革病毒免疫层析快速检测方法,简便、快捷、特异性好,可用于大规模快速诊断。

Objective To establish a novel method for the direct detection of dengue virus antigen by using nanomagnetic beads based on immunochromatography.Methods The nano-magnetic beads were coated with dengue virus labeling antibody to prepare immune-probe.The capture antibody for dengue virus was dispensed on nitrocellulose membrane as the test line,and goat anti-mouse polyclonal antibody was dispensed as the control line.The recombinant dengue virus NS1 protein was diluted to 100,50,20,10,1 and 0.1 ng/ml respectively,tested with the established nano-magnetic bead immunochromatographic test strips,and compared with the commercially available colloidal gold test strips.Results By observation,the detection limit of the nano-magnetic bead immunochromatographic test strips for detecting dengue virus NS1 protein was 1 ng/ml,and the detection limit of the commercially colloidal gold test strips was 10 ng/ml.After specific testing,there was no cross-reaction with West Nile virus,Zika virus pseudovirus,Ebola virus,and influenza A virus.Conclusion A rapid immunochromatographic detection method for dengue virus has been established,which was simple,fast,and specific,and can be used for large-scale rapid diagnosis.