The global pandemic caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)virus has necessitated rapid,easy-to-use,and accurate diagnostic methods to monitor the virus infection.Herein,a ratiometric flu...The global pandemic caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)virus has necessitated rapid,easy-to-use,and accurate diagnostic methods to monitor the virus infection.Herein,a ratiometric fluorescence enzyme-linked immunosorbent assay(ELISA)was developed using Si-fluorescein isothiocyanate nanoparticles(FITC NPs)for detecting SARSCoV-2 nucleocapsid(N)protein.Si-FITC NPs were prepared by a one-pot hydrothermal method using 3-aminopropyl triethoxysilane(APTES)-FITC as the Si source.This method did not need post-modification and avoided the reduction in quantum yield and stability.The p-nitrophenyl(pNP)produced by the alkaline phosphatase(ALP)-mediated hydrolysis of pnitrophenyl phosphate(pNPP)could quench Si fluorescence in Si-FITC NPs via the inner filter effect.In ELISA,an immunocomplex was formed by the recognition of capture antibody/N protein/reporter antibody.ALP-linked secondary antibody bound to the reporter antibody and induced pNPP hydrolysis to specifically quench Si fluorescence in Si-FITC NPs.The change in fluorescence intensity ratio could be used for detecting N protein,with a wide linearity range(0.01-10.0 and 50-300 ng/mL)and low detection limit(0.002 ng/mL).The concentration of spiked SARS-CoV-2 N protein could be determined accurately in human serum.Moreover,this proposed method can accurately distinguish coronavirus disease 2019(COVID-19)and non-COVID-19 patient samples.Therefore,this simple,sensitive,and accurate method can be applied for the early diagnosis of SARS-CoV-2 virus infection.展开更多
Cadmium ion(Cd^(2+))detection technology plays a prominent role in food safety and human health.Herein,we designed and constructed an 2-aminoethyl dihydrogen phosphate(AEP)@upconversion nanoparticles(UCNPs)fluorescenc...Cadmium ion(Cd^(2+))detection technology plays a prominent role in food safety and human health.Herein,we designed and constructed an 2-aminoethyl dihydrogen phosphate(AEP)@upconversion nanoparticles(UCNPs)fluorescence sensor for quantitative detection of Cd^(2+)in paddy rice based on inner filter effect(IFE)combined with enzyme inhibition mechanism.The AEP modification UCNPs can offer a stable fluorescence donor at 658 nm and be quenched by the oxidized tetramethylbenzidine(oxTMB)catalyzed by horseradish peroxidase(HRP)enzymes.Without addition of Cd^(2+),the fluorescence of AEP@UCNPs fluorescence sensor was weaken due to the IFE between AEP@UCNPs and oxTMB.With addition of Cd^(2+),HRP enzyme activity was inhibited by Cd^(2+),leading to the decreased oxTMB,resulting in the enhance upconversion fluorescence intensity.As a result,the fluorescence intensity signal at 658 nm of the IFE-based AEP@UCNPs fluorescence sensor increased linearly with the increase in Cd^(2+)in a wide range from 0.5μmol/L to 6μmol/L and the limit of detection(LOD)was 24.6 n mol/L.In addition,our proposed IFE-based AEP@UCNPs fluorescence sensor can achieve Cd^(2+)detection in paddy rice in 30 min.展开更多
基金supported by the National Key Research and Development Program of China(No.2021YFA0910900)the National Natural Science Foundation(No.22104147)+4 种基金Youth Innovation Promotion Association CAS(No.2021359)the Natural Science Foundation of Guangdong(Nos.2018B030306046 and 2020A1515111130)Guangdong Provincial Key Laboratory of Synthetic Genomics(No.2019B030301006)Shenzhen Science and Technology Program(No.KQTD20180413181837372)Shenzhen Outstanding Talents Training Fund.
文摘The global pandemic caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)virus has necessitated rapid,easy-to-use,and accurate diagnostic methods to monitor the virus infection.Herein,a ratiometric fluorescence enzyme-linked immunosorbent assay(ELISA)was developed using Si-fluorescein isothiocyanate nanoparticles(FITC NPs)for detecting SARSCoV-2 nucleocapsid(N)protein.Si-FITC NPs were prepared by a one-pot hydrothermal method using 3-aminopropyl triethoxysilane(APTES)-FITC as the Si source.This method did not need post-modification and avoided the reduction in quantum yield and stability.The p-nitrophenyl(pNP)produced by the alkaline phosphatase(ALP)-mediated hydrolysis of pnitrophenyl phosphate(pNPP)could quench Si fluorescence in Si-FITC NPs via the inner filter effect.In ELISA,an immunocomplex was formed by the recognition of capture antibody/N protein/reporter antibody.ALP-linked secondary antibody bound to the reporter antibody and induced pNPP hydrolysis to specifically quench Si fluorescence in Si-FITC NPs.The change in fluorescence intensity ratio could be used for detecting N protein,with a wide linearity range(0.01-10.0 and 50-300 ng/mL)and low detection limit(0.002 ng/mL).The concentration of spiked SARS-CoV-2 N protein could be determined accurately in human serum.Moreover,this proposed method can accurately distinguish coronavirus disease 2019(COVID-19)and non-COVID-19 patient samples.Therefore,this simple,sensitive,and accurate method can be applied for the early diagnosis of SARS-CoV-2 virus infection.
基金financially supported by the National Natural Science Foundation of China(32202132,32172229)Young Elite Scientists Sponsorship Program by CAST(2022QNRC001)the Priority Academic Program Development of Jiangsu Higher Educations(PAPD)。
文摘Cadmium ion(Cd^(2+))detection technology plays a prominent role in food safety and human health.Herein,we designed and constructed an 2-aminoethyl dihydrogen phosphate(AEP)@upconversion nanoparticles(UCNPs)fluorescence sensor for quantitative detection of Cd^(2+)in paddy rice based on inner filter effect(IFE)combined with enzyme inhibition mechanism.The AEP modification UCNPs can offer a stable fluorescence donor at 658 nm and be quenched by the oxidized tetramethylbenzidine(oxTMB)catalyzed by horseradish peroxidase(HRP)enzymes.Without addition of Cd^(2+),the fluorescence of AEP@UCNPs fluorescence sensor was weaken due to the IFE between AEP@UCNPs and oxTMB.With addition of Cd^(2+),HRP enzyme activity was inhibited by Cd^(2+),leading to the decreased oxTMB,resulting in the enhance upconversion fluorescence intensity.As a result,the fluorescence intensity signal at 658 nm of the IFE-based AEP@UCNPs fluorescence sensor increased linearly with the increase in Cd^(2+)in a wide range from 0.5μmol/L to 6μmol/L and the limit of detection(LOD)was 24.6 n mol/L.In addition,our proposed IFE-based AEP@UCNPs fluorescence sensor can achieve Cd^(2+)detection in paddy rice in 30 min.
