非包被ELISA可检测寨卡病毒包膜蛋白

A non?coated enzyme?linked immunosorbent assay for screening zika virus envelope protein

目的建立一种基于寨卡病毒包膜蛋白的非包被ELISA方法,检测被感染的细胞内E蛋白的表达情况,用于抗寨卡病毒药物筛选。方法将寨卡病毒感染的贴壁细胞固定在孔板上,加入抗体与之结合,并确定抗体浓度,建立E蛋白的非包被ELISA方法;用该方法检测木脂素类化合物C1的抗病毒活性;用实时荧光定量PCR实验(RT-PCR)验证这种非包被ELISA方法的准确性;并排除该方法对登革病毒的交叉反应。结果经过优化,未感染的细胞的背景A450nm降低到0.20左右,采用该方法检测的抗病毒活性结果,与实时荧光定量PCR实验的结果基本一致,且该方法对登革病毒不存在交叉反应。结论建立了一种寨卡病毒E蛋白的非包被ELISA方法,可用于抗寨卡病毒药物筛选。

Objective To establish a non-coated enzyme-linked immunosorbent assay (ELISA) based on zika virus envelope (E) protein for detecting the expression of E protein in infected cells.Methods Adherent Vero-143 cells infected with zika virus in a 96-well plate were fixed,and the antibodies against zika virus E protein were added at an optimized concentration to establish the non-coated ELISA method for E protein.The antiviral activities of lignans compound C1 was evaluated using this method.The accuracy of this non-coated ELISA was verified by RT-PCR,and the cross reaction with dengue virus was assessed.Results After optimization,the background absorbance at 450 nm of uninfected cells was reduced to about 0.20.The antiviral activities of lignans compound C1 detected by this method were basically consistent with the results of RT-PCR.No cross reaction with dengue virus was found in this assay.Conclusion A non- coated ELISA method based on zika virus E protein was established,which can be used for screening antiviral agents against zika virus.