Rapid and Sensitive Detection of Severe Acute Respiratory Syndrome Coronavirus 2 in Label-Free Manner Using Micromechanical Sensors

Coronavirus (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has been identified as a deadly pandemic. The genomic analysis of SARS-CoV-2 is performed using a reverse transcription-polymerase chain reaction (RT-PCR) technique for identifying viral ribonucleic acid (RNA) in infected patients. However, the RT-PCR diagnostic technique is manually laborious and expensive; therefore, it is not readily accessible in every laboratory. Methodological simplification is crucial to combat the ongoing pandemic by introducing quick, efficient, and affordable diagnostic methods. Here, we report how microcantilever sensors offer promising opportunities for rapid COVID-19 detection. Our first attempt was to capture the single-stranded complementary DNA of SARS-CoV-2 through DNA hybridization. Therefore, the microcantilever surface was immobilized with an oligonucleotide probe and detected using complementary target DNA hybridization by a shift in microcantilever resonance frequency. Our results show that microcantilever sensors can discriminate between complementary and noncomplementary target DNA on a micro to nanoscale. Additionally, the microcantilever sensors’ aptitude toward partial complementary DNA determines their potential to identify new variants of coronavirus. Therefore, microcantilever sensing could be a vital tool in the effort to extinguish the spreading COVID-19 pandemic.

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Label-Free Dengue Detection Utilizing PNA/DNA Hybridization Based on the Aggregation Process of Unmodified Gold Nanoparticles

Journal of Nanomaterials ◽

10.1155/2014/839286 ◽

2014 ◽

Vol 2014 ◽

pp. 1-5 ◽

Cited By ~ 5

Author(s):

Samsulida Abdul Rahman ◽

Rafidah Saadun ◽

Nur Ellina Azmi ◽

Nurhayati Ariffin ◽

Jaafar Abdullah ◽

...

Keyword(s):

Gold Nanoparticles ◽

Dna Hybridization ◽

Peptide Nucleic Acid ◽

Virus Detection ◽

Label Free ◽

Color Changes ◽

Phosphate Backbone ◽

Single Base Mismatch ◽

Target Dna ◽

Base Mismatch

A label-free optical detection method based on PNA/DNA hybridization using unmodified gold nanoparticles (AuNPs) for dengue virus detection has been successfully developed. In this study, no immobilization method is involved and the hybridization of PNA/DNA occurs directly in solution. Unmodified AuNPs undergo immediate aggregation in the presence of neutral charge peptide nucleic acid (PNA) due to the coating of PNA on AuNPs surface. However, in the presence of complementary targets DNA, the hybridization of PNA probe with target DNA forms negatively charged complexes due to the negatively charged phosphate backbone of the target DNA. The negatively charged complexes adsorbed onto the AuNPs surface ensure sufficient charge repulsion, need for AuNPs dispersion, and stability in solution. The detection procedure is a naked eye method based on immediate color changes and also through UV-vis adsorption spectra. The selectivity of the proposed method was studied successfully by single base mismatch and noncomplementary target DNA.

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Methods of Diagnosis a Preferential Advantage in COVID-19: A Mini Review

Journal of Pharmaceutical Research International ◽

10.9734/jpri/2021/v33i43a32476 ◽

2021 ◽

pp. 173-179

Author(s):

Neha Saini ◽

Prem Pandey ◽

Mandar Shirolkar ◽

Atul Kulkarni

Keyword(s):

Polymerase Chain Reaction ◽

Severe Acute Respiratory Syndrome ◽

Review Article ◽

Diagnostic Methods ◽

Rt Pcr ◽

Chain Reaction ◽

Health Crisis ◽

The World ◽

Polymerase Chain ◽

Novel Coronavirus

Humanity is going through never seen before health crisis due to the outbreak of novel coronavirus or Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2). There are 24.02 million cases and 0.82 million deaths worldwide as of 26th August 2020 due to deadly infection of COVID-19. The disease has been spreading exponentially (R-naught number: 3) and has challenged even the best healthcare infrastructure in the world. With the progression of the disease, the countries shifted the focus from cure to diagnosis and containment to flatten the curve. The review shows that the disease is spreading exponentially while the resources are still limited. We focus upon the probable vectors of the virus, different diagnostic methods with advantages & limitations, and the way forward. This review article covers the different diagnostic methods with more advantages, limitations, and the future sneak-peek into the forthcoming developments for the diagnostic processes such as RT-PCR (Reverse Transcription Polymerase chain reaction).

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Graphene-Based Hall Effect Biosensor for Improved Specificity and Sensitivity of Label-Free DNA Detection

NANO ◽

10.1142/s1793292020500885 ◽

2020 ◽

Vol 15 (07) ◽

pp. 2050088

Author(s):

Naiyuan Cui ◽

Fei Wang ◽

Hanyuan Ding

Keyword(s):

Hall Effect ◽

Dna Hybridization ◽

Chemical Vapor ◽

High Specificity ◽

Label Free ◽

Complementary Dna ◽

Mismatched Dna ◽

Specificity And Sensitivity ◽

Label Free Detection ◽

Detection Of Biomolecules

There is a broad interest in using graphene or graphene oxide (GO) sheets as a transducer for selective and label-free detection of biomolecules such as DNA, tumor marker, biological ions, etc. Here, a chemical vapor deposition (CVD) graphene-based Hall effect biosensor used for ultrasensitive label-free detection of DNA via DNA hybridization is reported. Hall effect measurements based on the Van der Pauw method are used to perform single-base sequence selective detection of DNA on graphene sheets, which are prepared by CVD. The mobility decreases and the sheet resistance increases with the adding of either complementary or one-base mismatched DNA to the graphene device. The hole carrier concentration of the graphene devices increases apparently with the addition of complementary DNA while it is hardly affected by the one-base mismatched DNA. The detection limit as low as 1[Formula: see text]pM was realized with a linear range from 1[Formula: see text]pM to 100[Formula: see text]nM. Moreover, the Hall effect biosensor was able to distinguish the complementary DNA from one-base mismatched DNA with a high specificity of [Formula: see text]6.2 which is almost two orders of magnitude higher than that of the previously reported graphene biosensors based on DNA–DNA hybridization.

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Label-Free Electrochemical Detection of DNA Hybridization Related to Anthrax Lethal Factor by using Carbon Nanotube Modified Sensors

Current Analytical Chemistry ◽

10.2174/1573411014666181004151547 ◽

2019 ◽

Vol 15 (4) ◽

pp. 502-510 ◽

Cited By ~ 1

Author(s):

Hakan Karadeniz ◽

Arzum Erdem

Keyword(s):

Detection Limit ◽

Dna Hybridization ◽

Lethal Factor ◽

Modified Electrodes ◽

Label Free ◽

Carbon Paste ◽

Chip Technology ◽

Anthrax Lethal Factor ◽

Hybridization Time ◽

Electrochemical Monitoring

Background: Anthrax Lethal Factor (ANT) is the dominant virulence factor produced by B. anthracis and is the major cause of death of infected animals. In this paper, pencil graphite electrodes GE were modified with single-walled and multi-walled carbon nanotubes (CNTs) for the detection of hybridization related to the ANT DNA for the first time in the literature. Methods: The electrochemical monitoring of label-free DNA hybridization related to ANT DNA was explored using both SCNT and MCNT modified PGEs with differential pulse voltammetry (DPV). The performance characteristics of ANT-DNA hybridization on disposable GEs were explored by measuring the guanine signal in terms of optimum analytical conditions; the concentration of SCNT and MCNT, the concentrations of probe and target, and also the hybridization time. Under the optimum conditions, the selectivity of probe modified electrodes was tested and the detection limit was calculated. Results: The selectivity of ANT probes immobilized onto MCNT-GEs was tested in the presence of hybridization of probe with NC no response was observed and with MM, smaller responses were observed in comparison to full-match DNA hybridization case. Even though there are unwanted substituents in the mixture samples containing both the target and NC in the ratio 1:1 and both the target and MM in the ratio 1:1, it has been found that ANT probe immobilized CNT modified graphite sensor can also select its target by resulting with 20.9% decreased response in comparison to the one measured in the case of full-match DNA hybridization case Therefore, it was concluded that the detection of direct DNA hybridization was performed by using MCNT-GEs with an acceptable selectivity. Conclusion: Disposable SCNT/MCNT modified GEs bring some important advantages to our assay including easy use, cost-effectiveness and giving a response in a shorter time compared to unmodified PGE, carbon paste electrode and glassy carbon electrode developed for electrochemical monitoring of DNA hybridization. Consequently, the detection of DNA hybridization related to the ANT DNA by MCNT modified sensors was performed by using lower CNT, probe and target concentrations, in a shorter hybridization time and resulting in a lower detection limit according to the SCNT modified sensors. In conclusion, MCNT modified sensors can yield the possibilities leading to the development of nucleic acid sensors platforms for the improvement of fast and cost-effective detection systems with respect to DNA chip technology.

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Proposal of RT-PCR–Based Mass Population Screening for Severe Acute Respiratory Syndrome Coronavirus 2 (Coronavirus Disease 2019)

Journal of Molecular Diagnostics ◽

10.1016/j.jmoldx.2020.07.001 ◽

2020 ◽

Vol 22 (10) ◽

pp. 1294-1299 ◽

Cited By ~ 3

Author(s):

Nikhil S. Sahajpal ◽

Ashis K. Mondal ◽

Allan Njau ◽

Sudha Ananth ◽

Kimya Jones ◽

...

Keyword(s):

Severe Acute Respiratory Syndrome ◽

Population Screening ◽

Rt Pcr

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Real-time label-free detection of DNA hybridization using a functionalized graphene field effect transistor: a theoretical study

Journal of Nanoparticle Research ◽

10.1007/s11051-021-05295-1 ◽

2021 ◽

Vol 23 (8) ◽

Author(s):

Sheida Bagherzadeh-Nobari ◽

Reza Kalantarinejad

Keyword(s):

Real Time ◽

Dna Hybridization ◽

Field Effect ◽

Field Effect Transistor ◽

Theoretical Study ◽

Functionalized Graphene ◽

Label Free ◽

Graphene Field Effect Transistor ◽

Label Free Detection ◽

Effect Transistor

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In Response to: Multiple assays in a real-time RT-PCR Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) panel can mitigate the risk of loss of sensitivity by new genomic variants during the COVID-19 outbreak

International Journal of Infectious Diseases ◽

10.1016/j.ijid.2021.01.049 ◽

2021 ◽

Author(s):

Luis Peñarrubia ◽

Maria Ruiz ◽

Roberto Porco ◽

Sonia N. Rao ◽

Stephen A. Vella ◽

...

Keyword(s):

Severe Acute Respiratory Syndrome ◽

Real Time ◽

Rt Pcr ◽

Genomic Variants

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CRISPR as a Diagnostic Tool

Biomolecules ◽

10.3390/biom11081162 ◽

2021 ◽

Vol 11 (8) ◽

pp. 1162

Author(s):

Seohyun Kim ◽

Sangmin Ji ◽

Hye Ran Koh

Keyword(s):

Diagnostic Tool ◽

Specific Binding ◽

Diagnostic Methods ◽

Specific Gene ◽

Guide Rna ◽

Engineering Technique ◽

Target Dna ◽

Target Binding ◽

Genetic Engineering Technique ◽

Disease Related Gene

Clustered regularly interspaced short palindromic repeats (CRISPR)-Cas system has recently gained growing attention as a diagnostic tool due to its capability of specific gene targeting. It consists of Cas enzymes and a guide RNA (gRNA) that can cleave the target DNA or RNA based on the sequence of the gRNA, making it an attractive genetic engineering technique. In addition to the target-specific binding and cleavage, the trans-cleavage activity was reported for some Cas proteins, including Cas12a and Cas13a, which is to cleave the surrounding single-stranded DNA or RNA upon the target binding of Cas-gRNA complex. All these activities of the CRISPR-Cas system are based on its target-specific binding, making it applied to develop diagnostic methods by detecting the disease-related gene as well as microRNAs and the genetic variations such as single nucleotide polymorphism and DNA methylation. Moreover, it can be applied to detect the non-nucleic acids target such as proteins. In this review, we cover the various CRISPR-based diagnostic methods by focusing on the activity of the CRISPR-Cas system and the form of the target. The CRISPR-based diagnostic methods without target amplification are also introduced briefly.

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