ABSTRACT
The SARS-CoV-2 spike glycoprotein (SARS-CoV-2-S) was used as a template molecule and polypyrrole (Ppy) was applied as an electro-generated conducting polymer, which was acting as a matrix for the formation of molecular imprints. Two types of Ppy-layers: molecularly imprinted polypyrrole (MIP-Ppy) and non-imprinted polypyrrole (NIP-Ppy) were electrochemically deposited on the working platinum electrode. The performance of electrodes modified by MIP-Ppy and NIP-Ppy layers was evaluated by pulsed amperometric detection (PAD). During the assessment of measurement results registered by PAD, the integrated Cottrell equation (Anson plot) was used to calculate the amount of charge passed through the MIP-Ppy and NIP-Ppy layers. The interaction between SARS-CoV-2 spike glycoproteins and molecularly imprinted polypyrrole (MIP-Ppy) was assessed by the Anson plot based calculations. This assessment reveals that SARS-CoV-2-S glycoproteins are interacting with MIP-Ppy more strongly than with NIP-Ppy. [Display omitted] • The SARS-CoV-2 spike glycoprotein (SARS-CoV-2-S) was molecularly imprinted within polypyrrole (Ppy). • Molecularly imprinted polypyrrole (MIP-Ppy) and non-imprinted polypyrrole (NIP-Ppy) were electro-deposited on Pt electrode. • Performance of electrodes modified by MIP-Ppy and NIP-Ppy was evaluated by pulsed amperometric detection (PAD). • Cottrell equation (Anson plot) was applied for the calculation of passed charge. • Interaction between SARS-CoV-2 protein and MIP-Ppy and NIP-Ppy was evaluated using Anson plot. [ FROM AUTHOR]
ABSTRACT
The SARS-CoV-2 spike glycoprotein (SARS-CoV-2-S) was used as a template molecule and polypyrrole (Ppy) was applied as an electro-generated conducting polymer, which was acting as a matrix for the formation of molecular imprints. Two types of Ppy-layers: molecularly imprinted polypyrrole (MIP-Ppy) and non-imprinted polypyrrole (NIP-Ppy) were electrochemically deposited on the working platinum electrode. The performance of electrodes modified by MIP-Ppy and NIP-Ppy layers was evaluated by pulsed amperometric detection (PAD). During the assessment of measurement results registered by PAD, the integrated Cottrell equation (Anson plot) was used to calculate the amount of charge passed through the MIP-Ppy and NIP-Ppy layers. The interaction between SARS-CoV-2 spike glycoproteins and molecularly imprinted polypyrrole (MIP-Ppy) was assessed by the Anson plot based calculations. This assessment reveals that SARS-CoV-2-S glycoproteins are interacting with MIP-Ppy more strongly than with NIP-Ppy.
ABSTRACT
The serologic diagnosis of coronavirus disease 2019 (COVID-19) and the evaluation of vaccination effectiveness are identified by the presence of antibodies specific to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In this paper, we present the electrochemical-based biosensing technique for the detection of antibodies specific to the SARS-CoV-2 proteins. Recombinant SARS-CoV-2 spike proteins (rSpike) were immobilised on the surface of a gold electrode modified by a self-assembled monolayer (SAM). This modified electrode was used as a sensitive element for the detection of polyclonal mouse antibodies against the rSpike (anti-rSpike). Electrochemical impedance spectroscopy (EIS) was used to observe the formation of immunocomplexes while cyclic voltammetry (CV) was used for additional analysis of the surface modifications. It was revealed that the impedimetric method and the elaborate experimental conditions are appropriate for the further development of electrochemical biosensors for the serological diagnosis of COVID-19 and/or the confirmation of successful vaccination against SARS-CoV-2.
Subject(s)
Biosensing Techniques , COVID-19 , Animals , Antibodies , Biosensing Techniques/methods , COVID-19/diagnosis , Electrochemical Techniques/methods , Humans , Mice , SARS-CoV-2 , Spike Glycoprotein, CoronavirusABSTRACT
Early detection of viral pathogens by DNA-sensors in clinical samples, contaminated foods, soil or water can dramatically improve clinical outcomes and reduce the socioeconomic impact of diseases such as COVID-19. Clustered regularly interspaced short palindromic repeat (CRISPR) and its associated protein Cas12a (previously known as CRISPR-Cpf1) technology is an innovative new-generation genomic engineering tool, also known as 'genetic scissors', that has demonstrated the accuracy and has recently been effectively applied as appropriate (E-CRISPR) DNA-sensor to detect the nucleic acid of interest. The CRISPR-Cas12a from Prevotella and Francisella 1 are guided by a short CRISPR RNA (gRNA). The unique simultaneous cis- and trans- DNA cleavage after target sequence recognition at the PAM site, sticky-end (5-7 bp) employment, and ssDNA/dsDNA hybrid cleavage strategies to manipulate the attractive nature of CRISPR-Cas12a are reviewed. DNA-sensors based on the CRISPR-Cas12a technology for rapid, robust, sensitive, inexpensive, and selective detection of virus DNA without additional sample purification, amplification, fluorescent-agent- and/or quencher-labeling are relevant and becoming increasingly important in industrial and medical applications. In addition, CRISPR-Cas12a system shows great potential in the field of E-CRISPR-based bioassay research technologies. Therefore, we are highlighting insights in this research direction.
Subject(s)
CRISPR-Cas Systems/physiology , DNA, Viral/isolation & purification , Nucleic Acid Amplification Techniques , Animals , Biosensing Techniques/methods , Biosensing Techniques/trends , COVID-19/virology , DNA, Viral/analysis , Environmental Pollutants/analysis , Environmental Pollutants/isolation & purification , Food Contamination/analysis , Humans , Molecular Typing/methods , Molecular Typing/trends , Nucleic Acid Amplification Techniques/methods , Nucleic Acid Amplification Techniques/trends , SARS-CoV-2/genetics , Virology/methods , Virology/trends , Virus Diseases/classification , Virus Diseases/diagnosis , Virus Diseases/virologyABSTRACT
Monitoring and tracking infection is required in order to reduce the spread of the coronavirus disease 2019 (COVID-19), induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). To achieve this goal, the development and deployment of quick, accurate, and sensitive diagnostic methods are necessary. The determination of the SARS-CoV-2 virus is performed by biosensing devices, which vary according to detection methods and the biomarkers which are inducing/providing an analytical signal. RNA hybridisation, antigen-antibody affinity interaction, and a variety of other biological reactions are commonly used to generate analytical signals that can be precisely detected using electrochemical, electrochemiluminescence, optical, and other methodologies and transducers. Electrochemical biosensors, in particular, correspond to the current trend of bioanalytical process acceleration and simplification. Immunosensors are based on the determination of antigen-antibody interaction, which on some occasions can be determined in a label-free mode with sufficient sensitivity.
Subject(s)
Biosensing Techniques/methods , COVID-19 Testing/methods , SARS-CoV-2/chemistry , Humans , Molecular Diagnostic Techniques , Nanostructures , SARS-CoV-2/isolation & purification , Serologic TestsABSTRACT
Viral infections are the most common among diseases that globally require around 60 percent of medical care. However, in the heat of the pandemic, there was a lack of medical equipment and inpatient facilities to provide all patients with viral infections. The detection of viral infections is possible in three general ways such as (i) direct virus detection, which is performed immediately 1-3 days after the infection, (ii) determination of antibodies against some virus proteins mainly observed during/after virus incubation period, (iii) detection of virus-induced disease when specific tissue changes in the organism. This review surveys some global pandemics from 1889 to 2020, virus types, which induced these pandemics, and symptoms of some viral diseases. Non-analytical methods such as radiology and microscopy also are overviewed. This review overlooks molecular analysis methods such as nucleic acid amplification, antibody-antigen complex determination, CRISPR-Cas system-based viral genome determination methods. Methods widely used in the certificated diagnostic laboratory for SARS-CoV-2, Influenza A, B, C, HIV, and other viruses during a viral pandemic are outlined. A comprehensive overview of molecular analytical methods has shown that the assay's sensitivity, accuracy, and suitability for virus detection depends on the choice of the number of regions in the viral open reading frame (ORF) genome sequence and the validity of the selected analytical method.