基于微波法快速合成胶体金建立猴痘病毒抗原免疫层析试纸条半定量方法

Establishment of a semi-quantitative immunochromatographic test strip approach based on a colloidal gold microwave method for detection of monkeypox viral antigen

目的 建立猴痘病毒(Monkeypox virus, MPXV)A35R蛋白现场快速半定量检测胶体金免疫层析方法。方法 以微波法6 min快速合成金纳米颗粒(Gold nanoparticles, AuNPs)作为标记物,基于双抗体夹心法体系建立检测MPXV A35R蛋白的免疫层析试纸方法。利用真核系统表达的MPXV包膜成分A35R蛋白配制系列浓度标准液,初步评价胶体金免疫层析试纸条的敏感性、特异性及稳定性等检测性能。结果 20 min内完成检测,建立MPXV抗原的胶体金免疫层析试纸的半定量检测的灵敏度达到62.5 pg/mL,肉眼最低检测限是125 pg/mL。与登革病毒(Dengue virus, DENV)、诺如病毒(Norovirus, NV)均无交叉反应,稳定性良好。结论 本研究建立的胶体金半定量免疫层析试纸条可以实现对MPXV的快速灵敏检测,在MPXV的即时检测(Point of care testing, POCT)领域具有一定潜力。

To establish an immunochromatographic method using colloidal gold for rapid,semi-quantitative,on-site detection of monkeypox virus(MPXV)A35R protein,in this research,gold nanoparticles(AuNPs)were synthesized through a microwave method within 6 minutes and selected for labeling,and an immunochromatographic test strip approach was developed for the detection of MPXV A35R protein with a double antibody sandwich system.A series of solutions of MPXV A35R protein,expressed via an eukaryotic expression system,were prepared to preliminarily evaluate the sensitivity,specificity,and stability of the colloidal gold immunoassay strip.The colloidal gold immunochromatographic strip detection of MPXV was completed within 20 minutes,the sensitivity of semi-quantitative detection reached 62.5 pg/mL,and the limit of detection by the naked eye was 125 pg/mL.No cross-reactivity with dengue virus(DENV)and norovirus(NV)was observed,and the stability was satisfactory.Thus,colloidal gold immunochromatography strips for semi-quantitative detection of MPXV were established in this research.This method may havepotential applications in point of care testing for MPXV.