摘要
目的 将内参蛋白引入新型冠状病毒(SARS-CoV-2)荧光免疫层析检测体系,以监测样本检测各环节的有效性,提高检测准确率。方法 聚乙烯亚胺(PEI)自组装法制备硅核双层量子点微球(SiO2-DQD),分别偶联内参抗体和SARS-CoV-2检测抗体。硝酸纤维素膜双检测线上分别包被内参抗体和SARS-CoV-2的捕获抗体。基于双抗夹心免疫检测原理,构建基于内参辅助的SARS-CoV-2荧光免疫层析检测方法。结果 荧光信号结果显示,在样本检测结果有效的条件下,该方法对SARS-CoV-2抗原的检测限可达0.01 ng/ml,且具有良好的特异性、稳定性。结论建立了快速的基于内参蛋白检测的SARS-CoV-2荧光免疫层析方法,为SARS-CoV-2抗原检测提供了质控对照,降低检测结果的假阴性率。
Objective To monitor the validity of the sample process flow and to improve the detection accuracy of a SARS⁃CoV⁃2antigenbyintroducing internalreferenceintoSARS⁃CoV⁃2fluorescentlateralflowassay.Methods Silica⁃core double⁃layer quantum dots(SiO2⁃DQD)microspheres were prepared using a polyethyleneimine(PEI)⁃mediated self⁃assembly method before being separately conjugated with an internal reference detection antibody and SARS⁃CoV⁃2 antibody.Their capture antibodies were plotted on two test lines of the nitrocellulose membrane.Based on double⁃antibody sandwich detection principles,an internal reference⁃assisted fluorescent lateral flow assay for SARS⁃CoV⁃2 antigen was established.Results According to the readout fluorescent signals,the detection limit of the method for the SARS⁃CoV⁃2 antigen reached 0.01 ng/ml with a validated sample process,suggesting its good specificity and stability.Conclusion In this study,a rapid internal reference⁃integrated fluorescent lateral flow assay for SARS⁃CoV⁃2 has been established,which provides control reference for the analysis workflow so that the false⁃negative rate of the test results can be reduced.
作者
王子仪
程小丹
杨兴胜
荣振
王升启
WANG Ziyi;CHENG Xiaodan;YANG Xingsheng;RONG Zhen;WANG Shengqi(Bioinformatics Center,Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 100850,China)
出处
《军事医学》
CAS
CSCD
2023年第12期913-918,共6页
Military Medical Sciences
基金
国家自然科学基金项目(81902159)。
关键词
新型冠状病毒
内参蛋白
免疫层析
量子点
荧光
SARS⁃CoV⁃2
internal reference protein
immunochromatography
quantum dots
fluorescence
