Abstract
We investigate electropolymerized molecularly imprinted polymers (E-MIPs) for the selective recognition of SARS-CoV-2 whole virus. E-MIPs imprinted with SARS-CoV-2 pseudoparticles (pps) were electrochemically deposited onto screen printed electrodes by reductive electropolymerization, using the water-soluble N-hydroxmethylacrylamide (NHMA) as functional monomer and crosslinked with N,N’-methylenebisacrylamide (MBAm). E-MIPs for SARS-CoV-2 showed selectivity for template SARS-CoV-2 pps, with an imprinting factor of 3:1, and specificity (significance = 0.06) when cross-reacted with other respiratory viruses. E-MIPs detected the presence of SARS-CoV-2 pps in <10 min with a limit of detection of 4.9 log 10 pfu/mL, suggesting their suitability for detection of SARS-CoV-2 with minimal sample preparation. Using electrochemical impedance spectroscopy (EIS) and principal component analysis (PCA), the capture of SARS-CoV-2 from real patient saliva samples was also evaluated. Fifteen confirmed COVID-19 positive and nine COVID-19 negative saliva samples were compared against the established loop-mediated isothermal nucleic acid amplification (LAMP) technique used by the UK National Health Service. EIS data demonstrated a PCA discrimination between positive and negative LAMP samples. A threshold real impedance signal (Z Re ) ≫ 4000 Ω and a corresponding charge transfer resistance (R CT ) ≫ 6000 Ω was indicative of absence of virus (COVID-19 negative) in agreement with values obtained for our control non-imprinted polymer control. A Z Re at or below a threshold value of 600 Ω with a corresponding R CT of <1200 Ω was indicative of a COVID-19 positive sample. The presence of virus was confirmed by treatment of E-MIPs with a SARS-CoV-2 specific monoclonal antibody.
Keywords: Biosensor; COVID-19; Electrochemical impedance spectroscopy; Electrochemical polymerization; Molecularly imprinted polymers; SARS-CoV-2.
【저자키워드】 COVID-19, SARS-CoV-2., biosensor, Electrochemical impedance spectroscopy, Electrochemical polymerization, Molecularly imprinted polymers, 【초록키워드】 Treatment, SARS-CoV-2, Saliva, monoclonal antibody, virus, principal component analysis, Polymer, respiratory viruses, biosensor, nucleic acid, specificity, LAMP, Patient, limit of detection, selectivity, respiratory, nucleic acid amplification, threshold, PCA, Other respiratory viruses, isothermal, capture, National Health Service, Principal component, transfer, saliva samples, monomer, threshold value, selective, positive, positive sample, N,N'-methylenebisacrylamide, UK National Health Service, log, evaluated, nine, functional, absence, demonstrated, COVID-19 negative, crosslinked, pseudoparticle, 【제목키워드】 SARS-CoV-2, Saliva, Polymer,