tetano
Editor, Senior Moderator
ACS Sens
. 2022 Mar 10.
doi: 10.1021/acssensors.1c02664. Online ahead of print.
Rapid, Point-of-Care scFv-SERS Assay for Femtogram Level Detection of SARS-CoV-2
Delphine Antoine[SUP] 1 [/SUP], Moein Mohammadi[SUP] 2 [/SUP], Madison Vitt[SUP] 2 [/SUP], Julia Marie Dickie[SUP] 2 [/SUP], Sharmin Sultana Jyoti[SUP] 2 [/SUP], Maura A Tilbury[SUP] 1 [/SUP], Patrick A Johnson[SUP] 2 [/SUP], Karen E Wawrousek[SUP] 2 [/SUP], J Gerard Wall[SUP] 1 [/SUP]
Affiliations
Abstract
Rapid, sensitive, on-site identification of SARS-CoV-2 infections is an important tool in the control and management of COVID-19. We have developed a surface-enhanced Raman scattering (SERS) immunoassay for highly sensitive detection of SARS-CoV-2. Single-chain Fv (scFv) recombinant antibody fragments that bind the SARS-CoV-2 spike protein were isolated by biopanning a human scFv library. ScFvs were conjugated to magnetic nanoparticles and SERS nanotags, followed by immunocomplex formation and detection of the SARS-CoV-2 spike protein with a limit of detection of 257 fg/mL in 30 min in viral transport medium. The assay also detected B.1.1.7 ("alpha"), B.1.351 ("beta"), and B.1.617.2 ("delta") spike proteins, while no cross-reactivity was observed with the common human coronavirus HKU1 spike protein. Inactivated whole SARS-CoV-2 virus was detected at 4.1 × 10[SUP]4[/SUP] genomes/mL, which was 10-100-fold lower than virus loads typical of infectious individuals. The assay exhibited higher sensitivity for SARS-CoV-2 than commercial lateral flow assays, was compatible with viral transport media and saliva, enabled rapid pivoting to detect new virus variants, and facilitated highly sensitive, point-of-care diagnosis of COVID-19 in clinical and public health settings.
Keywords: COVID-19; Raman spectroscopy; SARS-CoV-2; SERS; diagnostic; point-of-care; scFv antibody fragment.
. 2022 Mar 10.
doi: 10.1021/acssensors.1c02664. Online ahead of print.
Rapid, Point-of-Care scFv-SERS Assay for Femtogram Level Detection of SARS-CoV-2
Delphine Antoine[SUP] 1 [/SUP], Moein Mohammadi[SUP] 2 [/SUP], Madison Vitt[SUP] 2 [/SUP], Julia Marie Dickie[SUP] 2 [/SUP], Sharmin Sultana Jyoti[SUP] 2 [/SUP], Maura A Tilbury[SUP] 1 [/SUP], Patrick A Johnson[SUP] 2 [/SUP], Karen E Wawrousek[SUP] 2 [/SUP], J Gerard Wall[SUP] 1 [/SUP]
Affiliations
- PMID: 35271769
- DOI: 10.1021/acssensors.1c02664
Abstract
Rapid, sensitive, on-site identification of SARS-CoV-2 infections is an important tool in the control and management of COVID-19. We have developed a surface-enhanced Raman scattering (SERS) immunoassay for highly sensitive detection of SARS-CoV-2. Single-chain Fv (scFv) recombinant antibody fragments that bind the SARS-CoV-2 spike protein were isolated by biopanning a human scFv library. ScFvs were conjugated to magnetic nanoparticles and SERS nanotags, followed by immunocomplex formation and detection of the SARS-CoV-2 spike protein with a limit of detection of 257 fg/mL in 30 min in viral transport medium. The assay also detected B.1.1.7 ("alpha"), B.1.351 ("beta"), and B.1.617.2 ("delta") spike proteins, while no cross-reactivity was observed with the common human coronavirus HKU1 spike protein. Inactivated whole SARS-CoV-2 virus was detected at 4.1 × 10[SUP]4[/SUP] genomes/mL, which was 10-100-fold lower than virus loads typical of infectious individuals. The assay exhibited higher sensitivity for SARS-CoV-2 than commercial lateral flow assays, was compatible with viral transport media and saliva, enabled rapid pivoting to detect new virus variants, and facilitated highly sensitive, point-of-care diagnosis of COVID-19 in clinical and public health settings.
Keywords: COVID-19; Raman spectroscopy; SARS-CoV-2; SERS; diagnostic; point-of-care; scFv antibody fragment.