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1.
Viruses ; 12(12)2020 11 30.
Article in English | MEDLINE | ID: covidwho-1389520

ABSTRACT

Aptamers are short fragments of nucleic acids, DNA or RNA that have the ability to bind selected proteins with high specificity and affinity. These properties allow them to be used as an element of biosensors for the detection of specific proteins, including viral ones, which makes it possible to design valuable diagnostic tools. The influenza virus causes a huge number of human and animal deaths worldwide every year, and contributes to remarkable economic losses. In addition, in 2020, a new threat appeared-the SARS-Cov-2 pandemic. Both disease entities, especially in the initial stage of infection, are almost identical in terms of signs and symptoms. Therefore, a diagnostic solution is needed that will allow distinguishing between both pathogens, with high sensitivity and specificity; it should be cheap, quick and possible to use in the field, for example, in a doctor's office. All the mentioned properties are met by aptasensors in which the detection elements are specific aptamers. We present here the latest developments in the construction of various types of aptasensors for the detection of influenza virus. Aptasensor operation is based on the measurement of changes in electric impedance, fluorescence or electric signal (impedimetric, fluorescence and electrochemical aptasensors, respectively); it allows both qualitative and quantitative determinations. The particularly high advancement for detecting of influenza virus concerns impedimetric aptasensors.


Subject(s)
Aptamers, Nucleotide/therapeutic use , Biosensing Techniques , Influenza, Human/diagnosis , Orthomyxoviridae/isolation & purification , Aptamers, Nucleotide/genetics , COVID-19/diagnosis , Electric Impedance , Electrochemical Techniques , Fluorescence , Humans , SARS-CoV-2/isolation & purification
2.
J Hosp Infect ; 114: 144-152, 2021 Aug.
Article in English | MEDLINE | ID: covidwho-1343269

ABSTRACT

BACKGROUND: In a hospital setting, there is a need for rapid detection of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) to guide isolation measures and targeted admission. AIM: To evaluate the diagnostic performance of five SARS-CoV-2 rapid nucleocapsid protein antigen detection (RAD) assays (Biosynex, Biotical, Orient Gene, Panbio and SD Biosensor), and describe the performance and impact of implementation of the SD Biosensor assay in an emergency department. METHODS: Sensitivity and specificity of the five RAD assays were analysed on 100 respiratory samples: 60 real-time reverse transcriptase polymerase chain reaction (rRT-PCR)-confirmed SARS-CoV-2-positive samples, 24 SARS-CoV-2 RNA-negative samples and 16 samples positive for other respiratory pathogens. The manufacturer's protocol was adapted to validate the antigen tests on transport media used for rRT-PCR in the authors' routine practice. The SD Biosensor RAD assay was implemented as a screening method for rapid diagnosis and targeted admission. FINDINGS: Sensitivity of the five RAD assays ranged from 88.9% to 100% for samples with cycle threshold values <26, and specificity ranged from 46.2% to 100%. During the implementation period, 4195 RAD tests were performed. Due to the rapid RAD result, 157 patients were transferred directly to the coronavirus disease 2019 (COVID-19) cohort ward instead of the regular ward (N=47) or the temporary COVID-19 ward (N=110). CONCLUSION: The SD Biosensor, Biotical and Panbio SARS-CoV-2 antigen tests showed acceptable overall performance, and identified the majority of contagious patients. In the context of high prevalence of SARS-CoV-2, RAD tests can be used as a rapid screening tool to guide infection prevention measures and aid targeted admission.


Subject(s)
Antigens, Viral/isolation & purification , COVID-19 Testing , COVID-19 , COVID-19/diagnosis , COVID-19 Testing/methods , Hospitals , Humans , RNA, Viral , SARS-CoV-2 , Sensitivity and Specificity
3.
Lancet Infect Dis ; 21(8): 1089-1096, 2021 08.
Article in English | MEDLINE | ID: covidwho-1328820

ABSTRACT

BACKGROUND: Real-time PCR is recommended to detect SARS-CoV-2 infection. However, PCR availability is restricted in most countries. Rapid diagnostic tests are considered acceptable alternatives, but data are lacking on their performance. We assessed the performance of four antibody-based rapid diagnostic tests and one antigen-based rapid diagnostic test for detecting SARS-CoV-2 infection in the community in Cameroon. METHODS: In this clinical, prospective, diagnostic accuracy study, we enrolled individuals aged at least 21 years who were either symptomatic and suspected of having COVID-19 or asymptomatic and presented for screening. We tested peripheral blood for SARS-CoV-2 antibodies using the Innovita (Biological Technology; Beijing, China), Wondfo (Guangzhou Wondfo Biotech; Guangzhou, China), SD Biosensor (SD Biosensor; Gyeonggi-do, South Korea), and Runkun tests (Runkun Pharmaceutical; Hunan, China), and nasopharyngeal swabs for SARS-CoV-2 antigen using the SD Biosensor test. Antigen rapid diagnostic tests were compared with Abbott PCR testing (Abbott; Abbott Park, IL, USA), and antibody rapid diagnostic tests were compared with Biomerieux immunoassays (Biomerieux; Marcy l'Etoile, France). We retrospectively tested two diagnostic algorithms that incorporated rapid diagnostic tests for symptomatic and asymptomatic patients using simulation modelling. FINDINGS: 1195 participants were enrolled in the study. 347 (29%) tested SARS-CoV-2 PCR-positive, 223 (19%) rapid diagnostic test antigen-positive, and 478 (40%) rapid diagnostic test antibody-positive. Antigen-based rapid diagnostic test sensitivity was 80·0% (95% CI 71·0-88·0) in the first 7 days after symptom onset, but antibody-based rapid diagnostic tests had only 26·8% sensitivity (18·3-36·8). Antibody rapid diagnostic test sensitivity increased to 76·4% (70·1-82·0) 14 days after symptom onset. Among asymptomatic participants, the sensitivity of antigen-based and antibody-based rapid diagnostic tests were 37·0% (27·0-48·0) and 50·7% (42·2-59·1), respectively. Cohen's κ showed substantial agreement between Wondfo antibody rapid diagnostic test and gold-standard ELISA (κ=0·76; sensitivity 0·98) and between Biosensor and ELISA (κ=0·60; sensitivity 0·94). Innovita (κ=0·47; sensitivity 0·93) and Runkun (κ=0·43; sensitivity 0·76) showed moderate agreement. An antigen-based retrospective algorithm applied to symptomatic patients showed 94·0% sensitivity and 91·0% specificity in the first 7 days after symptom onset. For asymptomatic participants, the algorithm showed a sensitivity of 34% (95% CI 23·0-44·0) and a specificity of 92·0% (88·0-96·0). INTERPRETATION: Rapid diagnostic tests had good overall sensitivity for diagnosing SARS-CoV-2 infection. Rapid diagnostic tests could be incorporated into efficient testing algorithms as an alternative to PCR to decrease diagnostic delays and onward viral transmission. FUNDING: Médecins Sans Frontières WACA and Médecins Sans Frontières OCG. TRANSLATIONS: For the French and Spanish translations of the abstract see Supplementary Materials section.


Subject(s)
Antibodies, Viral/blood , Antigens, Viral/analysis , Asymptomatic Infections , COVID-19 Serological Testing , COVID-19/diagnosis , SARS-CoV-2/immunology , Feasibility Studies , Humans , Prospective Studies , Sensitivity and Specificity
4.
Anal Chim Acta ; 1159: 338384, 2021 May 15.
Article in English | MEDLINE | ID: covidwho-1279519

ABSTRACT

Viruses are the causing agents for many relevant diseases, including influenza, Ebola, HIV/AIDS, and COVID-19. Its rapid replication and high transmissibility can lead to serious consequences not only to the individual but also to collective health, causing deep economic impacts. In this scenario, diagnosis tools are of significant importance, allowing the rapid, precise, and low-cost testing of a substantial number of individuals. Currently, PCR-based techniques are the gold standard for the diagnosis of viral diseases. Although these allow the diagnosis of different illnesses with high precision, they still present significant drawbacks. Their main disadvantages include long periods for obtaining results and the need for specialized professionals and equipment, requiring the tests to be performed in research centers. In this scenario, biosensors have been presented as promising alternatives for the rapid, precise, low-cost, and on-site diagnosis of viral diseases. This critical review article describes the advancements achieved in the last five years regarding electrochemical biosensors for the diagnosis of viral infections. First, genosensors and aptasensors for the detection of virus and the diagnosis of viral diseases are presented in detail regarding probe immobilization approaches, detection methods (label-free and sandwich), and amplification strategies. Following, immunosensors are highlighted, including many different construction strategies such as label-free, sandwich, competitive, and lateral-flow assays. Then, biosensors for the detection of viral-diseases-related biomarkers are presented and discussed, as well as point of care systems and their advantages when compared to traditional techniques. Last, the difficulties of commercializing electrochemical devices are critically discussed in conjunction with future trends such as lab-on-a-chip and flexible sensors.


Subject(s)
Biosensing Techniques , Electrochemical Techniques , Virus Diseases/diagnosis , Viruses/isolation & purification , Humans , Immunoassay
5.
Biosens Bioelectron ; 190: 113421, 2021 Oct 15.
Article in English | MEDLINE | ID: covidwho-1260670

ABSTRACT

Early diagnosis and monitoring of SARS-CoV-2 virus is essential to control COVID-19 outbreak. In this study, we propose a promising surface enhanced Raman scattering (SERS)-based COVID-19 biosensor for ultrasensitive detection of SARS-CoV-2 virus in untreated saliva. The SERS-immune substrate was fabricated by a novel oil/water/oil (O/W/O) three-phase liquid-liquid interfaces self-assembly method, forming two layers of dense and uniform gold nanoparticle films to ensure the reproducibility and sensitivity of SERS immunoassay. The detection was performed by an immunoreaction between the SARS-CoV-2 spike antibody modified SERS-immune substrate, spike antigen protein and Raman reporter-labeled immuno-Ag nanoparticles. This SERS-based biosensor was able to detect the SARS-CoV-2 spike protein at concentrations of 0.77 fg mL-1 in phosphate-buffered saline and 6.07 fg mL-1 in untreated saliva. The designed SERS-based biosensor exhibited excellent specificity and sensitivity for SARS-CoV-2 virus without any sample pretreatment, providing a potential choice for the early diagnosis of COVID-19.


Subject(s)
Biosensing Techniques , COVID-19 , Metal Nanoparticles , Gold , Humans , Reproducibility of Results , SARS-CoV-2 , Saliva , Spike Glycoprotein, Coronavirus
6.
Biosens Bioelectron ; 190: 113418, 2021 Oct 15.
Article in English | MEDLINE | ID: covidwho-1260669

ABSTRACT

The continuing pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, which causes coronavirus disease 2019 (COVID-19), has spread globally and its reliable diagnosis is one of the foremost priorities for protecting public health. Herein a rapid (<1 h), easy-to-implement, and accurate CRISPR-based evanescent wave fluorescence biosensing platform for detection of SARS-CoV-2 is reported. The collateral effect of Cas13a is combined with a universal autonomous enzyme-free hybridization chain reaction (HCR) by designing a cleavage hairpin reporter, which is cleaved upon target recognition, and hence releasing the initiator sequence to trigger the downstream HCR circuits. Detection of HCR assemblies is accomplished by first adsorbing to the desthiobiotin-modified optical fiber, followed by fluorescence emission induced by an evanescent field. Three Cas13a crRNAs targeting the genes of S, N and Orf1ab of SARS-CoV-2 are programmed to specifically target SARS-CoV-2 or broadly detect related coronavirus strains, such as MERS-CoV and SARS-CoV. The HCR amplification coupled Cas13a-based biosensing platform is capable of rapid detection of SARS-CoV-2 with attomolar sensitivity. This method is further validated by adding target RNA of SARS-CoV-2 in negative oropharyngeal swabs. The good discrimination capability of this technique demonstrates its promising potential for point-of-care diagnosis of COVID-19.


Subject(s)
Biosensing Techniques , COVID-19 , CRISPR-Cas Systems , Clustered Regularly Interspaced Short Palindromic Repeats , Humans , Nucleic Acid Amplification Techniques , RNA, Viral , SARS-CoV-2 , Sensitivity and Specificity
7.
Anal Chem ; 93(24): 8585-8594, 2021 06 22.
Article in English | MEDLINE | ID: covidwho-1253862

ABSTRACT

The global pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus has revealed the urgent need for accurate, rapid, and affordable diagnostic tests for epidemic understanding and management by monitoring the population worldwide. Though current diagnostic methods including real-time polymerase chain reaction (RT-PCR) provide sensitive detection of SARS-CoV-2, they require relatively long processing time, equipped laboratory facilities, and highly skilled personnel. Laser-scribed graphene (LSG)-based biosensing platforms have gained enormous attention as miniaturized electrochemical systems, holding an enormous potential as point-of-care (POC) diagnostic tools. We describe here a miniaturized LSG-based electrochemical sensing scheme for coronavirus disease 2019 (COVID-19) diagnosis combined with three-dimensional (3D) gold nanostructures. This electrode was modified with the SARS-CoV-2 spike protein antibody following the proper surface modifications proved by X-ray photoelectron spectroscopy (XPS) and scanning electron microscopy (SEM) characterizations as well as electrochemical techniques. The system was integrated into a handheld POC detection system operated using a custom smartphone application, providing a user-friendly diagnostic platform due to its ease of operation, accessibility, and systematic data management. The analytical features of the electrochemical immunoassay were evaluated using the standard solution of S-protein in the range of 5.0-500 ng/mL with a detection limit of 2.9 ng/mL. A clinical study was carried out on 23 patient blood serum samples with successful COVID-19 diagnosis, compared to the commercial RT-PCR, antibody blood test, and enzyme-linked immunosorbent assay (ELISA) IgG and IgA test results. Our test provides faster results compared to commercial diagnostic tools and offers a promising alternative solution for next-generation POC applications.


Subject(s)
Biosensing Techniques , COVID-19 , Graphite , Point-of-Care Systems , Antibodies, Viral , COVID-19/diagnosis , COVID-19 Testing , Gold , Humans , Lasers , Nanostructures , SARS-CoV-2 , Sensitivity and Specificity , Spike Glycoprotein, Coronavirus
8.
Int J Environ Res Public Health ; 18(11)2021 05 25.
Article in English | MEDLINE | ID: covidwho-1244009

ABSTRACT

New methodologies based on the principle of "sewage epidemiology" have been successfully applied before in the detection of illegal drugs. The study describes the idea of early detection of a virus, e.g., SARS-CoV-2, in wastewater in order to focus on the area of virus occurrence and supplement the results obtained from clinical examination. By monitoring temporal variation in viral loads in wastewater in combination with other analysis, a virus outbreak can be detected and its spread can be suppressed early. The use of biosensors for virus detection also seems to be an interesting application. Biosensors are highly sensitive, selective, and portable and offer a way for fast analysis. This manuscript provides an overview of the current situation in the area of wastewater analysis, including genetic sequencing regarding viral detection and the technological solution of an early warning system for wastewater monitoring based on biosensors.


Subject(s)
COVID-19 , Wastewater-Based Epidemiological Monitoring , Humans , Mutation , SARS-CoV-2 , Sewage , Waste Water
9.
Anal Bioanal Chem ; 413(16): 4137-4159, 2021 Jul.
Article in English | MEDLINE | ID: covidwho-1233243

ABSTRACT

Coronavirus disease 2019 (COVID-19) pandemic is currently a serious global health threat. While conventional laboratory tests such as quantitative real-time polymerase chain reaction (qPCR), serology tests, and chest computerized tomography (CT) scan allow diagnosis of COVID-19, these tests are time-consuming and laborious, and are limited in resource-limited settings or developing countries. Point-of-care (POC) biosensors such as chip-based and paper-based biosensors are typically rapid, portable, cost-effective, and user-friendly, which can be used for COVID-19 in remote settings. The escalating demand for rapid diagnosis of COVID-19 presents a strong need for a timely and comprehensive review on the POC biosensors for COVID-19 that meet ASSURED criteria: Affordable, Sensitive, Specific, User-friendly, Rapid and Robust, Equipment-free, and Deliverable to end users. In the present review, we discuss the importance of rapid and early diagnosis of COVID-19 and pathogenesis of COVID-19 along with the key diagnostic biomarkers. We critically review the most recent advances in POC biosensors which show great promise for the detection of COVID-19 based on three main categories: chip-based biosensors, paper-based biosensors, and other biosensors. We subsequently discuss the key benefits of these biosensors and their use for the detection of antigen, antibody, and viral nucleic acids. The commercial POC biosensors for COVID-19 are critically compared. Finally, we discuss the key challenges and future perspectives of developing emerging POC biosensors for COVID-19. This review would be very useful for guiding strategies for developing and commercializing rapid POC tests to manage the spread of infections.Graphical abstract.


Subject(s)
Biosensing Techniques , COVID-19 Testing/methods , COVID-19/diagnosis , Point-of-Care Systems , Antibodies, Viral/analysis , Antigens, Viral/analysis , Biosensing Techniques/instrumentation , Biosensing Techniques/methods , COVID-19 Nucleic Acid Testing/methods , Humans , SARS-CoV-2/genetics
10.
Sensors (Basel) ; 21(10)2021 May 17.
Article in English | MEDLINE | ID: covidwho-1234803

ABSTRACT

In this paper, a highly sensitive graphene-based multiple-layer (BK7/Au/PtSe2/Graphene) coated surface plasmon resonance (SPR) biosensor is proposed for the rapid detection of the novel Coronavirus (COVID-19). The proposed sensor was modeled on the basis of the total internal reflection (TIR) technique for real-time detection of ligand-analyte immobilization in the sensing region. The refractive index (RI) of the sensing region is changed due to the interaction of different concentrations of the ligand-analyte, thus impacting surface plasmon polaritons (SPPs) excitation of the multi-layer sensor interface. The performance of the proposed sensor was numerically investigated by using the transfer matrix method (TMM) and the finite-difference time-domain (FDTD) method. The proposed SPR biosensor provides fast and accurate early-stage diagnosis of the COVID-19 virus, which is crucial in limiting the spread of the pandemic. In addition, the performance of the proposed sensor was investigated numerically with different ligand-analytes: (i) the monoclonal antibodies (mAbs) as ligand and the COVID-19 virus spike receptor-binding domain (RBD) as analyte, (ii) the virus spike RBD as ligand and the virus anti-spike protein (IgM, IgG) as analyte and (iii) the specific probe as ligand and the COVID-19 virus single-standard ribonucleic acid (RNA) as analyte. After the investigation, the sensitivity of the proposed sensor was found to provide 183.33°/refractive index unit (RIU) in SPR angle (θSPR) and 833.33THz/RIU in SPR frequency (SPRF) for detection of the COVID-19 virus spike RBD; the sensitivity obtained 153.85°/RIU in SPR angle and 726.50THz/RIU in SPRF for detection of the anti-spike protein, and finally, the sensitivity obtained 140.35°/RIU in SPR angle and 500THz/RIU in SPRF for detection of viral RNA. It was observed that whole virus spike RBD detection sensitivity is higher than that of the other two detection processes. Highly sensitive two-dimensional (2D) materials were used to achieve significant enhancement in the Goos-Hänchen (GH) shift detection sensitivity and plasmonic properties of the conventional SPR sensor. The proposed sensor successfully senses the COVID-19 virus and offers additional (1 + 0.55) × L times sensitivity owing to the added graphene layers. Besides, the performance of the proposed sensor was analyzed based on detection accuracy (DA), the figure of merit (FOM), signal-noise ratio (SNR), and quality factor (QF). Based on its performance analysis, it is expected that the proposed sensor may reduce lengthy procedures, false positive results, and clinical costs, compared to traditional sensors. The performance of the proposed sensor model was checked using the TMM algorithm and validated by the FDTD technique.


Subject(s)
Biosensing Techniques , COVID-19 , Graphite , Humans , SARS-CoV-2 , Surface Plasmon Resonance
11.
J Med Eng Technol ; 45(6): 423-433, 2021 Aug.
Article in English | MEDLINE | ID: covidwho-1230941

ABSTRACT

Acute respiratory syndrome coronavirus 2 (SARS-CoV-2), also called COVID-19, is one of the most contagious viruses resulting in a progressive pandemic. Since specific antiviral treatments have not been developed yet and its fatal rate is almost high, early and fast detection is critical for controlling the outbreak. In this study, a piezoelectric microcantilever biosensor has been designed for detecting COVID-19 samples directly without requiring preparation steps. The biosensor acts as a transducer and is coated with the related antibody. When the SARS-CoV-2 antigens adsorbed on the microcantilever top surface through their spike proteins, a surface stress due to the mass change would be prompted leading to the measurable tip deflection and floating voltage. To obtain a biosensor with optimum parameters, different shapes and piezoelectric materials have been assessed and it was concluded that a Poly (vinylidene fluoride) (PVDF) biosensor in a shape of a holed punched form triangle, represented the best result. Therefore, the highly sensitive microcantilever biosensor can detect COVID-19 in clinical samples with various viral loads, rapidly. Also, it is selective enough to differentiate SARS-CoV-2 from other viruses with similar symptoms.


Subject(s)
COVID-19/virology , Biosensing Techniques , Humans , Pandemics , Polymers/chemistry , SARS-CoV-2/genetics , SARS-CoV-2/pathogenicity
12.
Matter ; 4(7): 2403-2416, 2021 Jul 07.
Article in English | MEDLINE | ID: covidwho-1225332

ABSTRACT

SARS-CoV-2, the virus that causes COVID-19, has killed over 3 million people worldwide. Despite the urgency of the current pandemic, most available diagnostic methods for COVID-19 use RT-PCR to detect nucleic acid sequences specific to SARS-CoV-2. These tests are limited by their requirement of a large laboratory space, high reagent costs, multistep sample preparation, and the potential for cross-contamination. Moreover, results usually take hours to days to become available. Therefore, fast, reliable, inexpensive, and scalable point-of-care diagnostics are urgently needed. Here, we describe RAPID 1.0, a simple, handheld, and highly sensitive miniaturized biosensor modified with human receptor angiotensin-converting enzyme-2. RAPID 1.0 can detect SARS-CoV-2 using 10 µL of sample within 4 min through its increased resistance to charge transfer of a redox probe measured by electrochemical impedance spectroscopy. The sensitivity and specificity of RAPID for nasopharyngeal/oropharyngeal swab and saliva samples are 85.3% and 100% and 100% and 86.5%, respectively.

13.
Biosens Bioelectron ; 186: 113309, 2021 May 10.
Article in English | MEDLINE | ID: covidwho-1225153

ABSTRACT

The pandemic of coronavirus disease 2019 (COVID-19) caused by the severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) is continuously worsening globally, herein we have proposed an electrochemical biosensor for the sensitive monitoring of SARS-CoV-2 RNA. The presence of target RNA firstly triggers the catalytic hairpin assembly circuit and then initiates terminal deoxynucleotidyl transferase-mediated DNA polymerization. Consequently, a large number of long single-stranded DNA products can be produced, and these negatively charged DNA products will bind a massive of positively charged electroactive molecular of Ru(NH3)63+ due to the electrostatic adsorption. Therefore, significantly amplified electrochemical signals can be generated for sensitive analysis of SARS-CoV-2 RNA in the range of 0.1-1000 pM with the detection limit as low as 26 fM. Besides the excellent distinguishing ability for SARS-CoV-2 RNA against single-base mismatched RNA, the proposed biosensor can also be successfully applied to complex matrices, as well as clinical patient samples with high stability, which shows great prospects of clinical application.

14.
Nat Protoc ; 16(6): 3141-3162, 2021 06.
Article in English | MEDLINE | ID: covidwho-1209962

ABSTRACT

The global pandemic of coronavirus disease 2019 (COVID-19) highlights the shortcomings of the current testing paradigm for viral disease diagnostics. Here, we report a stepwise protocol for an RNA-extraction-free nano-amplified colorimetric test for rapid and naked-eye molecular diagnosis of COVID-19. The test employs a unique dual-prong approach that integrates nucleic acid (NA) amplification and plasmonic sensing for point-of-care detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), with a sample-to-assay response time of <1 h. The RNA-extraction-free nano-amplified colorimetric test utilizes plasmonic gold nanoparticles capped with antisense oligonucleotides (ASOs) as a colorimetric reporter to detect the amplified nucleic acid from the COVID-19 causative virus, SARS-CoV-2. The ASOs are specific for the SARS-CoV-2 N-gene, and binding of the ASOs to their target sequence results in the aggregation of the plasmonic gold nanoparticles. This highly specific agglomeration step leads to a change in the plasmonic response of the nanoparticles. Furthermore, when tested using clinical samples, the accuracy, sensitivity and specificity of the test were found to be >98.4%, >96.6% and 100%, respectively, with a detection limit of 10 copies/µL. The test can easily be adapted to diagnose other viral infections with a simple modification of the ASOs and primer sequences. It also provides a low-cost, rapid approach requiring minimal instrumentation that can be used as a screening tool for the diagnosis of COVID-19 at point-of-care settings in resource-poor situations. The colorimetric readout of the test can even be monitored using a handheld optical reader to obtain a quantitative response. Therefore, we anticipate that this protocol will be widely useful for the development of biosensors for the molecular diagnostics of COVID-19 and other infectious diseases.


Subject(s)
COVID-19 Nucleic Acid Testing/methods , COVID-19/diagnosis , Gold/chemistry , Metal Nanoparticles/chemistry , Oligonucleotides, Antisense/chemistry , RNA, Viral/analysis , SARS-CoV-2/isolation & purification , COVID-19/virology , COVID-19 Nucleic Acid Testing/instrumentation , Colorimetry/instrumentation , Colorimetry/methods , Humans , Limit of Detection , Oligonucleotides, Antisense/genetics , Point-of-Care Testing , RNA, Viral/genetics , SARS-CoV-2/genetics
15.
Biosens Bioelectron ; 185: 113177, 2021 Aug 01.
Article in English | MEDLINE | ID: covidwho-1206999

ABSTRACT

Rapid diagnosis and case isolation are pivotal to controlling the current pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In this study, a label-free DNA capacitive biosensor for the detection of SARS-CoV-2 that demonstrates real-time, low-cost, and high-throughput screening of nucleic acid samples is presented. Our novel biosensor composed of the interdigitated platinum/titanium electrodes on the glass substrate can detect the hybridization of analyte DNA with probe DNA. The hybridization signals of specific DNA sequences were verified through exhaustive physicochemical analytical techniques such as Fourier transform infrared (FT-IR) spectrometry, contact-angle analysis, and capacitance-frequency measurements. For a single-step hybridized reaction, the fabricated kit exhibited significant sensitivity (capacitance change, ΔC = ~2 nF) in detecting the conserved region of the SARS-CoV-2 RNA-dependent RNA polymerase (RdRp) gene with high sensitivity of 0.843 nF/nM. In addition to capacitive measurements, this selective detection was confirmed by the fluorescence image and intensity from a SARS-CoV-2 gene labeled with a fluorescent dye. We also demonstrated that the kits are recyclable by surface ozone treatment using UV irradiation. Thus, these kits could potentially be applied to various types of label-free DNA, thereby acting as rapid, cost-effective biosensors for several diseases.


Subject(s)
Biosensing Techniques , COVID-19 , DNA , Humans , Point-of-Care Systems , RNA, Viral , SARS-CoV-2 , Sensitivity and Specificity , Spectroscopy, Fourier Transform Infrared
16.
J Electroanal Chem (Lausanne) ; 893: 115289, 2021 Jul 15.
Article in English | MEDLINE | ID: covidwho-1201492

ABSTRACT

The SARS-CoV-2 virus is still causing a dramatic loss of human lives worldwide, constituting an unprecedented challenge for the society, public health and economy, to overcome. The up-to-date diagnostic tests, PCR, antibody ELISA and Rapid Antigen, require special equipment, hours of analysis and special staff. For this reason, many research groups have focused recently on the design and development of electrochemical biosensors for the SARS-CoV-2 detection, indicating that they can play a significant role in controlling COVID disease. In this review we thoroughly discuss the transducer electrode nanomaterials investigated in order to improve the sensitivity, specificity and response time of the as-developed SARS-CoV-2 electrochemical biosensors. Particularly, we mainly focus on the results appeard on Au-based and carbon or graphene-based electrodes, which are the main material groups recently investigated worldwidely. Additionally, the adopted electrochemical detection techniques are also discussed, highlighting their pros and cos. The nanomaterial-based electrochemical biosensors could enable a fast, accurate and without special cost, virus detection. However, further research is required in terms of new nanomaterials and synthesis strategies in order the SARS-CoV-2 electrochemical biosensors to be commercialized.

17.
ACS Appl Mater Interfaces ; 13(17): 19816-19824, 2021 May 05.
Article in English | MEDLINE | ID: covidwho-1199255

ABSTRACT

The detection of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is crucial for preventing and controlling infectious diseases and disease treatment. In this work, a Au@Ti3C2@PEI-Ru(dcbpy)32+ nanocomposite-based electrochemiluminescence (ECL) biosensor was rationally designed, which realized sensitive detection of the RNA-dependent RNA polymerase (RdRp) gene of SARS-CoV-2. In addition, a DNA walker was also used to excise the hairpin DNAs under the action of Nb.BbvCI endonuclease. Furthermore, model DNA-Ag nanoclusters (model DNA-AgNCs) were used to quench the initial ECL signal. As a result, the ECL biosensor was used to sensitively detect the SARS-CoV-2 RdRp gene with a detection range of 1 fM to 100 pM and a limit of detection of 0.21 fM. It was indicated that the ECL biosensor had a great application potential for clinical medical detection. Furthermore, the DNA walker amplification also played a reliable candidate strategy for other detection methods.


Subject(s)
Biosensing Techniques/methods , Nanocomposites/chemistry , SARS-CoV-2/genetics , DNA, Viral/genetics , RNA-Dependent RNA Polymerase/metabolism
18.
Molecules ; 25(20)2020 Oct 13.
Article in English | MEDLINE | ID: covidwho-1197552

ABSTRACT

The global SARS-CoV-2 pandemic started late 2019 and currently continues unabated. The lag-time for developing vaccines means it is of paramount importance to be able to quickly develop and repurpose therapeutic drugs. Protein-based biosensors allow screening to be performed using routine molecular laboratory equipment without a need for expensive chemical reagents. Here we present a biosensor for the 3-chymotrypsin-like cysteine protease from SARS-CoV-2, comprising a FRET-capable pair of fluorescent proteins held in proximity by a protease cleavable linker. We demonstrate the utility of this biosensor for inhibitor discovery by screening 1280 compounds from the Library of Pharmaceutically Active Compounds collection. The screening identified 65 inhibitors, with the 20 most active exhibiting sub-micromolar inhibition of 3CLpro in follow-up EC50 assays. The top hits included several compounds not previously identified as 3CLpro inhibitors, in particular five members of a family of aporphine alkaloids that offer promise as new antiviral drug leads.


Subject(s)
Betacoronavirus/drug effects , Biosensing Techniques/methods , Coronavirus Infections/drug therapy , Fluorescence Resonance Energy Transfer/methods , Pneumonia, Viral/drug therapy , Protease Inhibitors/chemistry , Protease Inhibitors/pharmacology , Viral Nonstructural Proteins/antagonists & inhibitors , Betacoronavirus/enzymology , Betacoronavirus/isolation & purification , COVID-19 , Coronavirus 3C Proteases , Coronavirus Infections/virology , Cysteine Endopeptidases , High-Throughput Screening Assays , Humans , Pandemics , Pneumonia, Viral/virology , SARS-CoV-2
19.
Biosensors (Basel) ; 11(4)2021 Apr 15.
Article in English | MEDLINE | ID: covidwho-1186891

ABSTRACT

The rapid spread of epidemic diseases (i.e., coronavirus disease 2019 (COVID-19)) has contributed to focus global attention on the diagnosis of medical conditions by ultrasensitive detection methods. To overcome this challenge, increasing efforts have been driven towards the development of single-molecule analytical platforms. In this context, recent progress in plasmonic biosensing has enabled the design of novel detection strategies capable of targeting individual molecules while evaluating their binding affinity and biological interactions. This review compiles the latest advances in plasmonic technologies for monitoring clinically relevant biomarkers at the single-molecule level. Functional applications are discussed according to plasmonic sensing modes based on either nanoapertures or nanoparticle approaches. A special focus was devoted to new analytical developments involving a wide variety of analytes (e.g., proteins, living cells, nucleic acids and viruses). The utility of plasmonic-based single-molecule analysis for personalized medicine, considering technological limitations and future prospects, is also overviewed.


Subject(s)
Biosensing Techniques/methods , Virus Diseases/diagnosis , Biomarkers/analysis , Biomarkers/metabolism , Biosensing Techniques/instrumentation , COVID-19/diagnosis , COVID-19/virology , Humans , Nanoparticles/chemistry , Nucleic Acids/analysis , SARS-CoV-2/genetics , SARS-CoV-2/isolation & purification , Single-Cell Analysis , Surface Plasmon Resonance , Virus Diseases/virology
20.
Sens Actuators A Phys ; 327: 112742, 2021 Aug 15.
Article in English | MEDLINE | ID: covidwho-1172076

ABSTRACT

Undoubtedly, the coronavirus disease 2019 (COVID-19) has received the greatest concern with a global impact, and this situation will continue for a long period of time. Looking back in history, airborne transimission diseases have caused huge casualties several times. COVID-19 as a typical airborne disease caught our attention and reminded us of the importance of preventing such diseases. Therefore, this study focuses on finding a new way to guard against the spread of these diseases such as COVID-19. This paper studies the dynamic electromechanical response of metal-core piezoelectric fiber/epoxy matrix composites, designed as mass load sensors for virus detection, by numerical modelling. The dynamic electromechanical response is simulated by applying an alternating current (AC) electric field to make the composite vibrate. Furthermore, both concentrated and distributed loads are considered to assess the sensitivity of the biosensor during modelling of the combination of both biomarker and viruses. The design parameters of this sensor, such as the resonant frequency, the position and size of the biomarker, will be studied and optimized as the key values to determine the sensitivity of detection. The novelty of this work is to propose functional composites that can detect the viruses from changes of the output voltage instead of the resonant frequency change using piezoelectric sensor and piezoelectric actuator. The contribution of this detection method will significantly shorten the detection time as it avoids fast Fourier transform (FFT) or discrete Fourier transform (DFT). The outcome of this research offers a reliable numerical model to optimize the design of the proposed biosensor for virus detection, which will contribute to the production of high-performance piezoelectric biosensors in the future.

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