tetano
Editor, Senior Moderator
Anal Chim Acta
. 2023 Oct 23:1279:341838.
doi: 10.1016/j.aca.2023.341838. Epub 2023 Sep 21. Combining hybrid nanoflowers with hybridization chain reaction for highly sensitive detection of SARS-CoV-2 nucleocapsid protein
Wen Yin[SUP] 1 [/SUP], Ji Hu[SUP] 2 [/SUP], Fang Chen[SUP] 2 [/SUP], Li Zhu[SUP] 2 [/SUP], Yingxin Ma[SUP] 3 [/SUP], Nuo Wang[SUP] 4 [/SUP], Hongping Wei[SUP] 5 [/SUP], Hang Yang[SUP] 5 [/SUP], Shan-Ho Chou[SUP] 2 [/SUP], Jin He[SUP] 6 [/SUP]
Affiliations
Background: COVID-19 (coronavirus disease 2019) pandemic has had enormous social and economic impacts so far. The nucleocapsid protein (N protein) is highly conserved and is a key antigenic marker for the diagnosis of early SARS-CoV-2 infection.
Results: In this study, the N protein was first captured by an aptamer (Aptamer 58) coupled to magnetic beads (MBs), which in turn were bound to another DNA sequence containing the aptamer (Aptamer 48-Initiator). After adding 5'-biotinylated hairpin DNA Amplifier 1 and Amplifier 2 with cohesive ends for complementary hybridization, the Initiator in the Aptamer 48-Initiator began to trigger the hybridization chain reaction (HCR), generating multiple biotin-labeled DNA concatamers. When incubated with synthetic streptavidin-invertase-Ca[SUB]3[/SUB](PO[SUB]4[/SUB])[SUB]2[/SUB] hybrid nanoflower (SICa), DNA concatamers could specifically bind to SICa through biotin-streptavidin interaction with high affinity. After adding sucrose, invertase in SICa hydrolyzed sucrose to glucose, whose concentration could be directly read with a portable glucometer, and its concentration was positively correlated with the amount of captured N protein. The method is highly sensitive with a detection limit as low as 1 pg/mL.
Significance: We believe this study provided a practical solution for the early detection of SARS-CoV-2 infection, and offered a new method for detecting other viruses through different target proteins.
Keywords: Aptamer; Hybrid nanoflower; Hybridization chain reaction (HCR); Nucleocapsid protein; Portable glucometer detection; SARS-CoV-2.
. 2023 Oct 23:1279:341838.
doi: 10.1016/j.aca.2023.341838. Epub 2023 Sep 21. Combining hybrid nanoflowers with hybridization chain reaction for highly sensitive detection of SARS-CoV-2 nucleocapsid protein
Wen Yin[SUP] 1 [/SUP], Ji Hu[SUP] 2 [/SUP], Fang Chen[SUP] 2 [/SUP], Li Zhu[SUP] 2 [/SUP], Yingxin Ma[SUP] 3 [/SUP], Nuo Wang[SUP] 4 [/SUP], Hongping Wei[SUP] 5 [/SUP], Hang Yang[SUP] 5 [/SUP], Shan-Ho Chou[SUP] 2 [/SUP], Jin He[SUP] 6 [/SUP]
Affiliations
- PMID: 37827653
- DOI: 10.1016/j.aca.2023.341838
Background: COVID-19 (coronavirus disease 2019) pandemic has had enormous social and economic impacts so far. The nucleocapsid protein (N protein) is highly conserved and is a key antigenic marker for the diagnosis of early SARS-CoV-2 infection.
Results: In this study, the N protein was first captured by an aptamer (Aptamer 58) coupled to magnetic beads (MBs), which in turn were bound to another DNA sequence containing the aptamer (Aptamer 48-Initiator). After adding 5'-biotinylated hairpin DNA Amplifier 1 and Amplifier 2 with cohesive ends for complementary hybridization, the Initiator in the Aptamer 48-Initiator began to trigger the hybridization chain reaction (HCR), generating multiple biotin-labeled DNA concatamers. When incubated with synthetic streptavidin-invertase-Ca[SUB]3[/SUB](PO[SUB]4[/SUB])[SUB]2[/SUB] hybrid nanoflower (SICa), DNA concatamers could specifically bind to SICa through biotin-streptavidin interaction with high affinity. After adding sucrose, invertase in SICa hydrolyzed sucrose to glucose, whose concentration could be directly read with a portable glucometer, and its concentration was positively correlated with the amount of captured N protein. The method is highly sensitive with a detection limit as low as 1 pg/mL.
Significance: We believe this study provided a practical solution for the early detection of SARS-CoV-2 infection, and offered a new method for detecting other viruses through different target proteins.
Keywords: Aptamer; Hybrid nanoflower; Hybridization chain reaction (HCR); Nucleocapsid protein; Portable glucometer detection; SARS-CoV-2.