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1.
ACS Appl Bio Mater ; 4(4): 2974-2995, 2021 04 19.
Article in English | MEDLINE | ID: covidwho-1157888

ABSTRACT

The current scenario, an ongoing pandemic of COVID-19, places a dreadful burden on the healthcare system worldwide. Subsequently, there is a need for a rapid, user-friendly, and inexpensive on-site monitoring system for diagnosis. The early and rapid diagnosis of SARS-CoV-2 plays an important role in combating the outbreak. Although conventional methods such as PCR, RT-PCR, and ELISA, etc., offer a gold-standard solution to manage the pandemic, they cannot be implemented as a point-of-care (POC) testing arrangement. Moreover, surface-enhanced Raman spectroscopy (SERS) having a high enhancement factor provides quantitative results with high specificity, sensitivity, and multiplex detection ability but lacks in POC setup. In contrast, POC devices such as lateral flow immunoassay (LFIA) offer rapid, simple-to-use, cost-effective, reliable platform. However, LFIA has limitations in quantitative and sensitive analyses of SARS-CoV-2 detection. To resolve these concerns, herein we discuss a unique modality that is an integration of SERS with LFIA for quantitative analyses of SARS-CoV-2. The miniaturization ability of SERS-based devices makes them promising in biosensor application and has the potential to make a better alternative of conventional diagnostic methods. This review also demonstrates the commercially available and FDA/ICMR approved LFIA kits for on-site diagnosis of SARS-CoV-2.


Subject(s)
COVID-19/diagnosis , Immunoassay/methods , Point-of-Care Systems , Spectrum Analysis, Raman , Viral Proteins/immunology , Antibodies, Immobilized/chemistry , Antibodies, Immobilized/immunology , Antibodies, Viral/blood , Antibodies, Viral/immunology , Biomarkers/blood , Biomarkers/metabolism , COVID-19/virology , Humans , SARS-CoV-2/isolation & purification , SARS-CoV-2/metabolism , Viral Proteins/metabolism
2.
Talanta ; 227: 122207, 2021 May 15.
Article in English | MEDLINE | ID: covidwho-1078201

ABSTRACT

Since December 2019, Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has caused millions of deaths and seriously threatened the safety of human life; indeed, this situation is worsening and many people are infected with the new coronavirus every day. Therefore, it is very important to understand patients' degree of infection and infection history through antibody testing. Such information is useful also for the government and hospitals to formulate reasonable prevention policies and treatment plans. In this paper, we develop a lateral flow immunoassay (LFIA) method based on superparamagnetic nanoparticles (SMNPs) and a giant magnetoresistance (GMR) sensing system for the simultaneously quantitative detection of anti-SARS-CoV-2 immunoglobulin M (IgM) and G (IgG). A simple and time-effective co-precipitation method was utilized to prepare the SMNPs, which have good dispersibility and magnetic property, with an average diameter of 68 nm. The Internet of Medical Things-supported GMR could transmit medical data to a smartphone through the Bluetooth protocol, making patient information available for medical staff. The proposed GMR system, based on SMNP-supported LFIA, has an outstanding advantage in cost-effectiveness and time-efficiency, and is easy to operate. We believe that the suggested GMR based LFIA system will be very useful for medical staff to analyze and to preserve as a record of infection in COVID-19 patients.


Subject(s)
Antibodies, Viral/blood , Immunoassay/methods , Immunoglobulin G/blood , Immunoglobulin M/blood , SARS-CoV-2/immunology , Animals , Antibodies, Immobilized/chemistry , Antibodies, Immobilized/immunology , Antibodies, Viral/immunology , Cattle , Cell Phone , Humans , Immunoglobulin G/immunology , Immunoglobulin M/immunology , Internet of Things , Limit of Detection , Magnetic Iron Oxide Nanoparticles/chemistry , Magnetic Phenomena
3.
ACS Appl Bio Mater ; 4(2): 1307-1318, 2021 02 15.
Article in English | MEDLINE | ID: covidwho-1069089

ABSTRACT

Recent evidence suggests that proinflammatory cytokines, such as tumor necrosis factor α (TNF-α), play a pivotal role in the development of inflammatory-related pathologies (covid-19, depressive disorders, sepsis, cancer, etc.,). More importantly, the development of TNF-α biosensors applied to biological fluids (e.g. sweat) could offer non-invasive solutions for the continuous monitoring of these disorders, in particular, polydimethylsiloxane (PDMS)-based biosensors. We have therefore investigated the biofunctionalization of PDMS surfaces using a silanization reaction with 3-aminopropyltriethoxysilane, for the development of a human TNF-α biosensor. The silanization conditions for 50 µm PDMS surfaces were extensively studied by using water contact angle measurements, electron dispersive X-ray and Fourier transform infrared spectroscopies, and fluorescamine detection. Evaluation of the wettability of the silanized surfaces and the Si/C ratio pointed out to the optimal silanization conditions supporting the formation of a stable and reproducible aminosilane layer, necessary for further bioconjugation. An ELISA-type immunoassay was then successfully performed for the detection and quantification of human TNF-α through fluorescent microscopy, reaching a limit of detection of 0.55 µg/mL (31.6 nM). Finally, this study reports for the first time a promising method for the development of PDMS-based biosensors for the detection of TNF-α, using a quick, stable, and simple biofunctionalization process.


Subject(s)
Dimethylpolysiloxanes/chemistry , Immunoassay/methods , Tumor Necrosis Factor-alpha/analysis , Antibodies, Immobilized/chemistry , Antibodies, Immobilized/immunology , COVID-19/metabolism , COVID-19/pathology , COVID-19/virology , Carbon/chemistry , Humans , Immunoassay/instrumentation , Limit of Detection , Microfluidics , Microscopy, Fluorescence , SARS-CoV-2/isolation & purification , Silicon/chemistry , Tumor Necrosis Factor-alpha/immunology , Wettability
4.
Adv Mater ; 33(10): e2007847, 2021 Mar.
Article in English | MEDLINE | ID: covidwho-1062095

ABSTRACT

The graphene revolution, which has taken place during the last 15 years, has represented a paradigm shift for science. The extraordinary properties possessed by this unique material have paved the road to a number of applications in materials science, optoelectronics, energy, and sensing. Graphene-related materials (GRMs) are now produced in large scale and have found niche applications also in the biomedical technologies, defining new standards for drug delivery and biosensing. Such advances position GRMs as novel tools to fight against the current COVID-19 and future pandemics. In this regard, GRMs can play a major role in sensing, as an active component in antiviral surfaces or in virucidal formulations. Herein, the most promising strategies reported in the literature on the use of GRM-based materials against the COVID-19 pandemic and other types of viruses are showcased, with a strong focus on the impact of functionalization, deposition techniques, and integration into devices and surface coatings.


Subject(s)
COVID-19/diagnosis , Graphite/chemistry , Nanostructures/chemistry , Antibodies, Immobilized/chemistry , Antibodies, Immobilized/immunology , Antiviral Agents/chemistry , Antiviral Agents/pharmacology , Biosensing Techniques/methods , COVID-19/prevention & control , COVID-19/virology , Electrochemical Techniques , Electrodes , Humans , Limit of Detection , Nanostructures/toxicity , SARS-CoV-2/drug effects , SARS-CoV-2/isolation & purification , SARS-CoV-2/metabolism , Surface Properties , Viral Proteins/analysis , Viral Proteins/immunology
5.
Anal Chim Acta ; 1147: 30-37, 2021 Feb 22.
Article in English | MEDLINE | ID: covidwho-1012278

ABSTRACT

Simple, low-cost, and sensitive new platforms for electrochemical immunosensors for virus detection have been attracted attention due to the recent pandemic caused by a new type of coronavirus (SARS-CoV-2). In the present work, we report for the first time the construction of an immunosensor using a commercial 3D conductive filament of carbon black and polylactic acid (PLA) to detect Hantavirus Araucaria nucleoprotein (Np) as a proof-of-concept. The recognition biomolecule was anchored directly at the filament surface by using N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride and N-Hydroxysuccinimide (EDC/NHS). Conductive and non-conductive composites of PLA were characterized using thermal gravimetric analysis (TGA), revealing around 30% w/w of carbon in the filament. Morphological features of composites were obtained from SEM and TEM measurements. FTIR measurement revealed that crosslinking agents were covalently bonded at the filament surface. Electrochemical techniques such as cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were used for the evaluation of each step involved in the construction of the proposed immunosensor. The results showed the potentiality of the device for the quantitative detection of Hantavirus Araucaria nucleoprotein (Np) from 30 µg mL-1 to 240 µg mL-1 with a limit of detection of 22 µg mL-1. Also, the proposed immunosensor was applied with success for virus detection in 100x diluted human serum samples. Therefore, the PLA conductive filament with carbon black is a simple and excellent platform for immunosensing, which offers naturally carboxylic groups able to anchor covalently biomolecules.


Subject(s)
Antibodies, Viral/immunology , Immunoassay/methods , Nucleocapsid Proteins/immunology , Printing, Three-Dimensional , Antibodies, Immobilized/chemistry , Antibodies, Immobilized/immunology , COVID-19/diagnosis , COVID-19/virology , Dielectric Spectroscopy , Electrodes , Hantavirus/isolation & purification , Hantavirus/metabolism , Hantavirus Infections/diagnosis , Hantavirus Infections/virology , Humans , Immunoassay/instrumentation , Limit of Detection , Nucleocapsid Proteins/blood , SARS-CoV-2/isolation & purification , Soot/chemistry
6.
Biosens Bioelectron ; 177: 112971, 2021 Apr 01.
Article in English | MEDLINE | ID: covidwho-1009326

ABSTRACT

Effective and efficient management of human betacoronavirus severe acute respiratory syndrome (SARS)-CoV-2 virus infection i.e., COVID-19 pandemic, required sensitive and selective sensors with short sample-to-result durations for performing desired diagnostics. In this direction, one appropriate alternative approach to detect SARS-CoV-2 virus protein at low level i.e., femtomolar (fM) is exploring plasmonic metasensor technology for COVID-19 diagnostics, which offers exquisite opportunities in advanced healthcare programs, and modern clinical diagnostics. The intrinsic merits of plasmonic metasensors stem from their capability to squeeze electromagnetic fields, simultaneously in frequency, time, and space. However, the detection of low-molecular weight biomolecules at low densities is a typical drawback of conventional metasensors that has recently been addressed using toroidal metasurface technology. This research is focused on the fabrication of a miniaturized plasmonic immunosensor based on toroidal electrodynamics concept that can sustain robustly confined plasmonic modes with ultranarrow lineshapes in the terahertz (THz) frequencies. By exciting toroidal dipole mode using our quasi-infinite metasurface and a judiciously optimized protocol based on functionalized gold nanoparticles (AuNPs) conjugated with the specific monoclonal antibody specific to spike protein (S1) of SARS-CoV-2 virus onto the metasurface, the resonance shifts for diverse concentrations of the spike protein are monitored. Possessing molecular weight around ~76 kDa allowed to detect the presence of SARS-CoV-2 virus protein with significantly low as limit of detection (LoD) was achieved as ~4.2 fM. We envisage that outcomes of this research will pave the way toward the use of toroidal metasensors as practical technologies for rapid and precise screening of SARS-CoV-2 virus carriers, symptomatic or asymptomatic, and spike proteins in hospitals, clinics, laboratories, and site of infection.


Subject(s)
Antibodies, Immobilized/chemistry , Biosensing Techniques/methods , COVID-19 Serological Testing/methods , COVID-19/diagnosis , SARS-CoV-2/isolation & purification , Spike Glycoprotein, Coronavirus/analysis , COVID-19/virology , Gold/chemistry , Humans , Immunoassay/methods , Limit of Detection , Metal Nanoparticles/chemistry
7.
Biosens Bioelectron ; 171: 112685, 2021 Jan 01.
Article in English | MEDLINE | ID: covidwho-891295

ABSTRACT

The spread of SARS-CoV-2 virus in the ongoing global pandemic has led to infections of millions of people and losses of many lives. The rapid, accurate and convenient SARS-CoV-2 virus detection is crucial for controlling and stopping the pandemic. Diagnosis of patients in the early stage infection are so far limited to viral nucleic acid or antigen detection in human nasopharyngeal swab or saliva samples. Here we developed a method for rapid and direct optical measurement of SARS-CoV-2 virus particles in one step nearly without any sample preparation using a spike protein specific nanoplasmonic resonance sensor. As low as 370 vp/mL were detected in one step within 15 min and the virus concentration can be quantified linearly in the range of 0 to 107 vp/mL. Measurements shown on both generic microplate reader and a handheld smartphone connected device suggest that our low-cost and rapid detection method may be adopted quickly under both regular clinical environment and resource-limited settings.


Subject(s)
Betacoronavirus/isolation & purification , Biosensing Techniques/instrumentation , Clinical Laboratory Techniques , Coronavirus Infections/diagnosis , Pneumonia, Viral/diagnosis , Point-of-Care Testing , Virion/isolation & purification , Antibodies, Immobilized/chemistry , Biosensing Techniques/economics , COVID-19 , COVID-19 Testing , Clinical Laboratory Techniques/economics , Coronavirus Infections/economics , Equipment Design , Humans , Limit of Detection , Models, Molecular , Pandemics , SARS-CoV-2 , Spike Glycoprotein, Coronavirus/analysis , Time Factors
8.
Sci Adv ; 6(42)2020 10.
Article in English | MEDLINE | ID: covidwho-781066

ABSTRACT

To combat severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) and any unknown emerging pathogens in the future, the development of a rapid and effective method to generate high-affinity antibodies or antibody-like proteins is of critical importance. We here report high-speed in vitro selection of multiple high-affinity antibody-like proteins against various targets including the SARS-CoV-2 spike protein. The sequences of monobodies against the SARS-CoV-2 spike protein were successfully procured within only 4 days. Furthermore, the obtained monobody efficiently captured SARS-CoV-2 particles from the nasal swab samples of patients and exhibited a high neutralizing activity against SARS-CoV-2 infection (half-maximal inhibitory concentration, 0.5 nanomolar). High-speed in vitro selection of antibody-like proteins is a promising method for rapid development of a detection method for, and of a neutralizing protein against, a virus responsible for an ongoing, and possibly a future, pandemic.


Subject(s)
Betacoronavirus/immunology , Peptidyl-Dipeptidase A/immunology , Single-Domain Antibodies/immunology , Spike Glycoprotein, Coronavirus/immunology , Amino Acid Sequence , Angiotensin-Converting Enzyme 2 , Antibodies, Immobilized/chemistry , Antibodies, Immobilized/immunology , Antibodies, Neutralizing/chemistry , Antibodies, Neutralizing/immunology , Antibodies, Neutralizing/metabolism , Betacoronavirus/genetics , Betacoronavirus/isolation & purification , COVID-19 , Cell Surface Display Techniques/methods , Coronavirus Infections/pathology , Coronavirus Infections/virology , Dimerization , Humans , Kinetics , Pandemics , Peptides/chemistry , Peptides/immunology , Peptidyl-Dipeptidase A/chemistry , Peptidyl-Dipeptidase A/metabolism , Pneumonia, Viral/pathology , Pneumonia, Viral/virology , Protein Domains/immunology , Protein Subunits/chemistry , Protein Subunits/immunology , Protein Subunits/metabolism , RNA, Viral/metabolism , SARS-CoV-2 , Single-Domain Antibodies/chemistry , Single-Domain Antibodies/metabolism , Spike Glycoprotein, Coronavirus/chemistry
9.
ACS Sens ; 5(9): 2747-2752, 2020 09 25.
Article in English | MEDLINE | ID: covidwho-740029

ABSTRACT

With the current intense need for rapid and accurate detection of viruses due to COVID-19, we report on a platform technology that is well suited for this purpose, using intact measles virus for a demonstration. Cases of infection due to the measles virus are rapidly increasing, yet current diagnostic tools used to monitor for the virus rely on slow (>1 h) technologies. Here, we demonstrate the first biosensor capable of detecting the measles virus in minutes with no preprocessing steps. The key sensing element is an electrode coated with a self-assembled monolayer containing the measles antibody, immobilized through an N-heterocyclic carbene (NHC). The intact virus is detected by changes in resistance, giving a linear response to 10-100 µg/mL of the intact measles virus without the need to label or process the sample. The limit of detection is 6 µg/mL, which is at the lower limit of concentrations that can cause infections in primates. The NHC-based biosensors are shown to be superior to thiol-based systems, producing an approximately 10× larger response and significantly greater stability toward repeated measurements and long-term storage. This NHC-based biosensor thus represents an important development for both the rapid detection of the measles virus and as a platform technology for the detection of other biological targets of interest.


Subject(s)
Antibodies, Immobilized/immunology , Benzimidazoles/chemistry , Biosensing Techniques/methods , Electrochemical Techniques/methods , Measles virus/isolation & purification , Antibodies, Immobilized/chemistry , Electrochemical Techniques/instrumentation , Electrodes , Gold/chemistry , Limit of Detection , Measles virus/immunology
10.
Biosens Bioelectron ; 165: 112361, 2020 Oct 01.
Article in English | MEDLINE | ID: covidwho-617487

ABSTRACT

The recent outbreak of the coronavirus disease (COVID-19) has left the world clueless. As the WHO declares this new contagion as a pandemic on the 11th of March 2020, the alarming rate of the spawn of the disease in such a short period has disarranged the globe. Standing against this situation researchers are strenuously searching for the key traits responsible for this pandemic. As knowledge regarding the dynamics and host-path interaction of COVID-19 causing Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) is currently unknown, the formulation of strategies concerning antiviral treatment, vaccination, and epidemiological control stands crucial. Before designing adequate therapeutic strategies, it is extremely essential to diagnose the disease at the outset as early detection can have a greater impact on building health system capacity. Hence, a comprehensive review of strategies for COVID-19 diagnosis is essential in this existing global situation. In this review, sequentially, we have provided the clinical details along with genetic and proteomic biomarkers related to COVID-19. The article systematically enlightens a clear overview of the clinically adopted techniques for the detection of COVID-19 including oligonucleotide-based molecular detection, Point-of-Care immunodiagnostics, radiographical analysis/sensing system, and newly developed biosensing prototypes having commercial viability. The commercial kits/analytical methods based-sensing strategies have also been tabulated categorically. The critical insights on the developer, commercial brand name, detection methods, technical operational details, detection time, clinical specimen, status, the limit of detection/detection ability have been discussed comprehensively. We believe that this review may provide scientists, clinicians and healthcare manufacturers valuable information regarding the most recent developments/approaches towards COVID-19 diagnosis.


Subject(s)
Betacoronavirus/isolation & purification , Biosensing Techniques/methods , Coronavirus Infections/diagnosis , Lab-On-A-Chip Devices , Pneumonia, Viral/diagnosis , Point-of-Care Testing , Animals , Antibodies, Immobilized/chemistry , Betacoronavirus/genetics , Biomarkers/analysis , Biosensing Techniques/instrumentation , COVID-19 , Clustered Regularly Interspaced Short Palindromic Repeats , Coronavirus Infections/blood , Coronavirus Infections/virology , Equipment Design , Humans , Immunoassay/instrumentation , Immunoassay/methods , Nanostructures/chemistry , Pandemics , Pneumonia, Viral/blood , Pneumonia, Viral/virology , Radiographic Image Enhancement/instrumentation , Radiographic Image Enhancement/methods , Reverse Transcriptase Polymerase Chain Reaction/instrumentation , Reverse Transcriptase Polymerase Chain Reaction/methods , SARS-CoV-2
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