Ratiometric fluorescent Si-FITC nanoprobe for immunoassay of SARS-CoV-2 nucleocapsid protein

Mao, Guobin1,2; Ye, Silu1,2,3; Yin, Wen3; Yang, Yang4; Ji, Xinghu5; He, Jin3; Liu, Yingxia4; Dai, Junbiao1,2; He, Zhike5; Ma, Yingxin1,2

2022-09-01

10.1007/s12274-022-5005-z

Nano Research   Impact Factor & Quartile

1998-0124

1998-0000

Coronavirus disease 2019 (COVID-19) highlights the importance of rapid and reliable diagnostic assays for the management of virus transmission. Here, we developed a one-pot hydrothermal method to prepare Si-FITC nanoparticles (NPs) for the fluorescent immunoassay of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleocapsid protein (N protein). The synthesis of Si-FITC NPs did not need post-modification, which addressed the issue of quantum yield reduction during the coupling reaction. Si-FITC NPs showed two distinct peaks, Si fluorescence at lambda(em) = 385 nm and FITC fluorescence at lambda(em) = 490 nm. In the presence of KMnO4, Si fluorescence was decreased and FITC fluorescence was enhanced. Briefly, in the presence of N protein, catalase (CAT)-linked secondary antibody/reporter antibody/N protein/capture antibody immunocomplexes were formed on microplates. Subsequently, hydrogen peroxide (H2O2) and Si-FITC NPs/KMnO4 were injected into the microplate together. The decomposition of H2O2 by CAT resulted in remaining of KMnO4, which changed the fluorescence intensity ratio of Si-FITC NPs. The fluorescence intensity ratio correlated significantly with the N protein concentration ranging from 0.02 to 50.00 ng/mL, and the detection limit was 0.003 ng/mL, which was more sensitive than the commercial ELISA kit with a detection limit of 0.057 ng/mL. The N protein concentration can be accurately determined in human serum. Furthermore, the COVID-19 and non-COVID-19 patients were distinguishable by this method. Therefore, the ratiometric fluorescent immunoassay can be used for SARS-CoV-2 infection diagnosis with a high sensitivity and selectivity.

Si-FITC nanoparticles ratiometric fluorescent probe SARS-CoV-2 ELISA

SCI

English

Others

National Key Research and Development Program of China[2021YFA0910900] ; Sino-German rapid response funding[C-0008] ; National Natural Science Foundation of China["32222044","22104147"] ; Shenzhen Municipal Science and Technology Innovation Council[RCYX20210609103823046] ; 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:000862228500004

TSINGHUA UNIV PRESS

Web of Science

1.Chinese Acad Sci, Guangdong Prov Key Lab Synthet Genom, CAS Key Lab Quantitat Engn Biol, Shenzhen Inst Adv Technol,Shenzhen Inst Synthet B, 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.Huazhong Agr Univ, Coll Life Sci & Technol, State Key Lab Agr Microbiol, Wuhan 430070, Peoples R China
4.Southern Univ Sci & Technol, Shenzhen Key Lab Pathogen & Immun, Shenzhen Peoples Hosp 3,Hosp 2, Natl Clin Res Ctr Infect Dis,State Key Discipline, Shenzhen 518112, Peoples R China
5.Wuhan Univ, Coll Chem & Mol Sci, Wuhan 430072, Peoples R China

Mao, Guobin,Ye, Silu,Yin, Wen,et al. Ratiometric fluorescent Si-FITC nanoprobe for immunoassay of SARS-CoV-2 nucleocapsid protein[J]. Nano Research,2022.

Mao, Guobin.,Ye, Silu.,Yin, Wen.,Yang, Yang.,Ji, Xinghu.,...&Ma, Yingxin.(2022).Ratiometric fluorescent Si-FITC nanoprobe for immunoassay of SARS-CoV-2 nucleocapsid protein.Nano Research.

Mao, Guobin,et al."Ratiometric fluorescent Si-FITC nanoprobe for immunoassay of SARS-CoV-2 nucleocapsid protein".Nano Research (2022).