Ratiometric fluorescence immunoassay of SARS-CoV-2 nucleocapsid protein via Si-FITC nanoprobe-based inner filter effect

Mao, Guobin1,2; Yang, Yang4; Cao, Shijie1,2,3; Ye, Silu1,2; Li, Yifang1,2; Zhao, Wei1,2; An, Hongwei3; Liu, Yingxia4; Dai, Junbiao1,2; Ma, Yingxin1,2

2022-08-01

10.1007/s12274-022-4740-5

Nano Research   影响因子和分区

1998-0124

1998-0000

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 SARS-CoV-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 p-nitrophenyl 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.

Si-fluorescein isothiocyanate (FITC) nanoparticles ratiometric fluorescent probe severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) inner filter effect enzyme-linked immunosorbent assay (ELISA)

SCI

英语

通讯

National Key Research and Development Program of China[2021YFA0910900] ; National Natural Science Foundation[22104147] ; Youth Innovation Promotion Association CAS[2021359] ; Natural Science Foundation of Guangdong["2018B030306046","2020A1515111130"] ; Guangdong Provincial Key Laboratory of Synthetic Genomics[2019B030301006] ; Shenzhen Science and Technology Program[KQTD20180413181837372]

Chemistry ; Science & Technology - Other Topics ; Materials Science ; Physics

Chemistry, Physical ; Nanoscience & Nanotechnology ; Materials Science, Multidisciplinary ; Physics, Applied

WOS:000842592900003

TSINGHUA UNIV PRESS

Web of Science

1.Chinese Acad Sci, Shenzhen Inst Synthet Biol, Guangdong Prov Key Lab Synthet Genom, CAS Key Lab Quantitat Engn Biol,Shenzhen Inst Adv, Shenzhen 518055, Peoples R China
2.Chinese Acad Sci, Shenzhen Key Lab Synthet Genom, Shenzhen Inst Synthet Biol, Shenzhen Inst Adv Technol, Shenzhen 518055, Peoples R China
3.Guangxi Univ Chinese Med, Nanning 530001, Peoples R China
4.Southern Univ Sci & Technol, Shenzhen Key Lab Pathogen & Immun, Natl Clin Res Ctr Infect Dis,Hosp 2, State Key Discipline Infect Dis,Shenzhen Peoples, Shenzhen 518112, Peoples R China

Mao, Guobin,Yang, Yang,Cao, Shijie,et al. Ratiometric fluorescence immunoassay of SARS-CoV-2 nucleocapsid protein via Si-FITC nanoprobe-based inner filter effect[J]. Nano Research,2022,16(4).

Mao, Guobin.,Yang, Yang.,Cao, Shijie.,Ye, Silu.,Li, Yifang.,...&Ma, Yingxin.(2022).Ratiometric fluorescence immunoassay of SARS-CoV-2 nucleocapsid protein via Si-FITC nanoprobe-based inner filter effect.Nano Research,16(4).

Mao, Guobin,et al."Ratiometric fluorescence immunoassay of SARS-CoV-2 nucleocapsid protein via Si-FITC nanoprobe-based inner filter effect".Nano Research 16.4(2022).