Coronavirus Anatomy and Its Analytical Approaches for Targeting COVID-19.

The whole world has lived in fear of the Coronavirus 2019 (COVID-19) for more than one-fourth of a year in 2020. Since then, numerous studies have been done and others are still being conducted to identify the coronavirus causal agent and determine the ideal antiviral therapies for treating this illness. Despite the fact of significant global efforts, which are being made to develop vaccines and identify therapeutic medicines, the emergence of multiple variants has rumbled the research for developing an ideal diagnostic and therapeutic tool for targeting COVID-19. A thorough anatomy of the coronavirus virus and its global dissemination is essential for healthcare initiatives, as well as for predicting and preventing new epidemics. Testing for active COVID-19 infections is a critical public health strategy for tracking the epidemic. The widespread adoption of polymerase chain reaction (PCR) and antigen rapid test assays has increased the supply of test kits. In this chapter, the significance of molecular technique to develop prognostic analytical techniques for targeting COVID-19 along with the advantages and limitations of available techniques is discussed.

Tailoring molecularly imprinted polymer on titanium-multiwalled carbon nanotube functionalized gold electrode for enhanced chlorophyll determination in microalgae health assessment.

A unique method for determining chlorophyll content in microalgae is devised employing a gold interdigitated electrode (G-IDE) with a 10-µm gap, augmented by a nano-molecularly imprinted polymer (nano-MIP) and a titanium dioxide/multiwalled carbon nanotube (TiO2/MWCNT) nanocomposite. The nano-MIP, produced using chlorophyll template voids, successfully trapped chlorophyll, while the TiO2/MWCNT nanocomposite, synthesized by the sol-gel technique, exhibited a consistent distribution and anatase crystalline structure. The rebinding of procured chlorophyll powder, which was used as a template for nano-MIP synthesis, was identified with a high determination coefficient (R2 = 0.9857). By combining the TiO2/MWCNT nanocomposite with nano-MIP, the G-IDE sensing method achieved a slightly better R2 value of 0.9892 for detecting chlorophyll in microalgae. The presented G-IDE sensor showed a significant threefold enhancement in chlorophyll detection compared with commercially available chlorophyll powder. It had a detection limit of 0.917 mL (v/v) and a linear range that spanned from 10-6 to 1 mL. The effectiveness of the sensor in detecting chlorophyll in microalgae was confirmed through validation of its repeatability and reusability.

Nanodiamond conjugated SARS-CoV-2 spike protein: electrochemical impedance immunosensing on a gold microelectrode.

A promising immunosensing strategy in diagnosing SARS-CoV-2 is proposed using a 10-µm gap-sized gold interdigitated electrode (AuIDE) to target the surface spike protein (SP). The microelectrode surface was modified by (3-glycidyloxypropyl) trimethoxysilane to enforce the epoxy matrix, which facilitates the immobilization of the anti-SP antibody. The immunosensing performance was evaluated by integrating a nanosized (~ 10 nm) diamond-complexed SP as a target. The proposed immunoassay was quantitatively evaluated through electrochemical impedance spectroscopy (EIS) with the swept frequency from 0.1 to 1 MHz using a 100 mVRMS AC voltage supply. The immunoassay performed without diamond integration showed low sensitivity, with the lowest SP concentration measured at 1 pM at a determination coefficient of R2 = 0.9681. In contrast, the nanodiamond-conjugated SP on the immunosensor showed excellent sensitivity with a determination coefficient of R2 = 0.986. SP detection with a nanodiamond-conjugated target on AuIDE reached the low limit of detection at 189 fM in a linear detection range from 250 to 8000 fM. The specificity of the developed immunosensor was evaluated by interacting influenza-hemagglutinin and SARS-CoV-2-nucleocapsid protein with anti-SP. In addition, the authentic interaction of SP and anti-SP was validated by enzyme-linked immunosorbent assay.

Molecularly imprinted polymer amalgamation on narrow-gapped Archimedean-spiral interdigitated electrodes: resistance to electrolyte fouling in acidic medium.

A conventional photolithography technique was used to fabricate three types of Archimedean-spiral interdigitated electrodes (AIDEs) containing concentric interlocking electrodes with different electrode and gap sizes, i.e., 150 µm (D1), 100 µm (D2), and 50 µm (D3). The precision of the fabrication was validated by surface topography using scanning electron microscopy, high power microscopy, 3D-nano profilometry, and atomic force microscopy. These AIDEs were fabricated with a tolerance of ± 6 nm in dimensions. The insignificant current variation at the pico-ampere range for all bare AIDEs further proved the reproducibility of the device. The large gap sized AIDE (D1) is insensitive to acidic medium, whereas D2 and D3 are insensitive to alkali medium. D2 was the best with regard to its electrical characterization. Furthermore, uniformly synthesized molecularly imprinted polymer (MIP) nanoparticles prepared with human blood clotting factor IX and its aptamer were in the size range 140 to 160 nm, attached on the sensing surface and characterized. The average thickness of deposited MIP film was 1.7 µm. EDX data shows the prominent peaks for silicon and aluminum substrates as 61.79 and 22.52%, respectively. The MIP nanoparticles-deposited sensor surface was characterized by applying it in electrolyte solutions, and smooth curves with the current flow were observed at pH lower than 8 and discriminated against alkali media. This study provides a new MIP amalgamated AIDE with nano-gapped fingers enabling analysis of other biomaterials due to its operation in an ideal buffer range.

A DNA based visual and colorimetric aggregation assay for the early growth factor receptor (EGFR) mutation by using unmodified gold nanoparticles.

A genomic DNA-based colorimetric assay is described for the detection of the early growth factor receptor (EGFR) mutation, which is the protruding reason for non-small cell lung cancer. A DNA sequence was designed and immobilized on unmodified gold nanoparticles (GNPs). The formation of the respective duplex indicates the presence of an EGFR mutation. It is accompanied by the aggregation of the GNPs in the presence of monovalent ions, and it indicates the presence of an EGFR mutation. This is accompanied by a color change from red (520 nm) to purple (620 nm). Aggregation was evidenced by transmission electron microscopy, scanning electron microscopy and atomic force microscopy. The limit of detection is 313 nM of the mutant target strand. A similar peak shift was observed for 2.5 µM concentrations of wild type target. No significant peak shift was observed with probe and non-complementary DNA. Graphical abstract Schematic representation of high-specific genomic DNA sequence on gold nanoparticle (GNP) aggregation with sodium chloride (NaCl). It illustrates the detection method for EGFR mutation on lung cancer detection. Red and purple colors of tubes represent dispersed and aggregated GNP, respectively.

A Review on Graphene Analytical Sensors for Biomarker-based Detection of Cancer.

The engineering of nanoscale materials has broadened the scope of nanotechnology in a restricted functional system. Today, significant priority is given to immediate health diagnosis and monitoring tools for point-of-care testing and patient care. Graphene, as a one-atom carbon compound, has the potential to detect cancer biomarkers and its derivatives. The atom-wide graphene layer specialises in physicochemical characteristics, such as improved electrical and thermal conductivity, optical transparency, and increased chemical and mechanical strength, thus making it the best material for cancer biomarker detection. The outstanding mechanical, electrical, electrochemical, and optical properties of two-dimensional graphene can fulfil the scientific goal of any biosensor development, which is to develop a more compact and portable point-of-care device for quick and early cancer diagnosis. The bio-functionalisation of recognised biomarkers can be improved by oxygenated graphene layers and their composites. The significance of graphene that gleans its missing data for its high expertise to be evaluated, including the variety in surface modification and analytical reports. This review provides critical insights into graphene to inspire research that would address the current and remaining hurdles in cancer diagnosis.

Transistor-Based Biomolecule Sensors: Recent Technological Advancements and Future Prospects.

Transistor-based sensors have been widely recognized to be highly sensitive and reliable for point-of-care/bed-side diagnosis. In this line, a range of cutting-edge technologies has been generated to elevate the role of transistors for biomolecule detection. Detection of a wide range of clinical biomarkers has been reported using various configurations of transistors. The inordinate sensitivity of transistors to the field-effect imparts high sensitivity toward wide range of biomolecules. This overview has gleaned the present achievements with the technological advancements using high performance transistor-based sensors. This review encloses transistors incorporated with a variety of functional nanomaterials and organic elements for their excellence in selectivity and sensitivity. In addition, the technological advancements in fabrication of these microdevices or nanodevices and functionalization of the sensing elements have also been discussed. The technological gap in the realization of sensors in transistor platforms and the resulted scope for research has been discussed. Finally, foreseen technological advancements and future research perspectives are described.

Non-protein coding RNA sequences mediate specific colorimetric detection of Staphylococcus aureus on unmodified gold nanoparticles.

Nonprotein coding RNA (npcRNA) is a transcribed gene sequence that is not able to translate into protein, yet it executes a specific function in modulation and regulation mechanisms. As npcRNA is highly resistant to the mutation, the Sau-02 npcRNA gene and its probe oligonucleotide, which are specifically present in Staphylococcus aureus and in methicillin-resistant S. aureus only, used to develop a highly specific and sensitive colorimetric assay on unmodified gold nanoparticles (AuNPs). Hybridization between the npcRNA Sau-02 gene sequences was detected through noncrosslinking AuNP aggregation in salt solution in the presence of probe-target gene sequences. AuNPs of 10 and 15 nm in sizes with monovalent ion salt (NaCl) solution were optimized as the ideal tool for investigating the stability of AuNPs upon the addition of gene sequences. The state dispersed and aggregated forms of 10 nm AuNPs with the presented colorimetric assay were justified through field emission scanning electron microscopy and atomic force microscopy. The particle distribution of two different AuNP states was evaluated through particle distribution analysis. The lowest detection amount of S. aureus npcRNA from the colorimetric assay performed was 6 pg/µL, as the color of AuNPs turned blue with the presence of probe oligonucleotides and target gene sequences.

Aptasensing nucleocapsid protein on nanodiamond assembled gold interdigitated electrodes for impedimetric SARS-CoV-2 infectious disease assessment.

In an aim of developing portable biosensor for SARS-CoV-2 pandemic, which facilitates the point-of-care aptasensing, a strategy using 10 µm gap-sized gold interdigitated electrode (AuIDE) is presented. The silane-modified AuIDE surface was deposited with ∼20 nm diamond and enhanced the detection of SARS-CoV-2 nucleocapsid protein (NCP). The characteristics of chemically modified diamond were evidenced by structural analyses, revealing the cubic crystalline nature at (220) and (111) planes as observed by XRD. XPS analysis denotes a strong interaction of carbon element, composed ∼95% as seen in EDS analysis. The C-C, CC, CO, CN functional groups were well-refuted from XPS spectra of carbon and oxygen elements in diamond. The interrelation between elements through FTIR analysis indicates major intrinsic bondings at 2687-2031 cm-1. The aptasensing was evaluated through electrochemical impedance spectroscopy measurements, using NCP spiked human serum. With a good selectivity the lower detection limit was evidenced as 0.389 fM, at a linear detection range from 1 fM to 100 pM. The stability, and reusability of the aptasensor were demonstrated, showing ∼30% and ∼33% loss of active state, respectively, after ∼11 days. The detection of NCP was evaluated by comparing anti-NCP aptamer and antibody as the bioprobes. The determination coefficients of R2 = 0.9759 and R2 = 0.9772 were obtained for aptamer- and antibody-based sensing, respectively. Moreover, the genuine interaction of NCP aptamer and protein was validated by enzyme linked apta-sorbent assay. The aptasensing strategy proposed with AuIDE/diamond enhanced sensing platform is highly recommended for early diagnosis of SARS-CoV-2 infection.

Graphitic carbon nitride/graphene nanoflakes hybrid system for electrochemical sensing of DNA bases in meat samples.

This research presents a simple, fast and simultaneous electrochemical quantitative determination of nucleobases, for example guanine (G), adenine (A), and thymine (T) in a beef and chicken livers samples to measure the quality of food products based on hybrids of graphitic carbon nitride/Graphene nanoflakes (g-C3N4/GNF) modified electrode. Graphitic carbon nitride (g-C3N4) made of graphite-like covalent link connects nitrogen, nitride, and carbon atoms in the structural design with improved the electrical properties and low band gap semiconductor. The g-C3N4/GNF nanocomposite was synthesized by the hydrothermal treatment to form a porous g-C3N4 interconnected three dimensional (3D) network of g-C3N4 and GNF. The 3D g-C3N4/GNF/GCE was utilized for the detection of nucleic acid bases with a well resolved oxidation peak for the individual analyte. The electrocatalytic current was established to be a linear range from 0.3 × 10-7 to 6.6 × 10-6, 0.3 × 10-7 to 7.3 × 10-6, and 5.3 × 10-6 to 63.3 × 10-4 M for G, A, and T with a detection limit of 4.7, 3.5 and 55 nM, respectively. The diffusion co-efficient and the kinetic parameters were derived from the chronoamperometry technique. The proposed sensing strategy has been effectively used for the application in real sample analysis and observed that the electrode free from the surface fouling.

Aluminosilicate Nanocomposites from Incinerated Chinese Holy Joss Fly Ash: A Potential Nanocarrier for Drug Cargos.

An incredible amount of joss fly ash is produced from the burning of Chinese holy joss paper; thus, an excellent method of recycling joss fly ash waste to extract aluminosilicate nanocomposites is explored. The present research aims to introduce a novel method to recycle joss fly ash through a simple and straightforward experimental procedure involving acidic and alkaline treatments. The synthesized aluminosilicate nanocomposite was characterized to justify its structural and physiochemical characteristics. A morphological analysis was performed with field-emission transmission electron microscopy, and scanning electron microscopy revealed the size of the aluminosilicate nanocomposite to be ~25 nm, while also confirming a uniformly spherical-shaped nanostructure. The elemental composition was measured by energy dispersive spectroscopy and revealed the Si to Al ratio to be 13.24 to 7.96, showing the high purity of the extracted nanocomposite. The roughness and particle distribution were analyzed using atomic force microscopy and a zeta analysis. X-ray diffraction patterns showed a synthesis of faceted and cubic aluminosilicate crystals in the nanocomposites. The presence of silica and aluminum was further proven by X-ray photoelectron spectroscopy, and the functional groups were recognized through Fourier transform infrared spectroscopy. The thermal capacity of the nanocomposite was examined by a thermogravimetric analysis. In addition, the research suggested the promising application of aluminosilicate nanocomposites as drug carriers. The above was justified by an enzyme-linked apta-sorbent assay, which claimed that the limit of the aptasensing aluminosilicate-conjugated ampicillin was two-fold higher than that in the absence of the nanocomposite. The drug delivery property was further justified through an antibacterial analysis against Escherichia coli (gram-negative) and Bacillus subtilis (gram-positive).

Multidimensional (0D-3D) nanostructures for lung cancer biomarker analysis: Comprehensive assessment on current diagnostics.

The pragmatic outcome of a lung cancer diagnosis is closely interrelated in reducing the number of fatal death caused by the world's top cancerous disease. Regardless of the advancement made in understanding lung tumor, and its multimodal treatment, in general the percentage of survival remain low. Late diagnosis of a cancerous cell in patients is the major hurdle for the above circumstances. In the new era of a lung cancer diagnosis with low cost, portable and non-invasive clinical sampling, nanotechnology is at its inflection point where current researches focus on the implementation of biosensor conjugated nanomaterials for the generation of the ideal sensing. The present review encloses the superiority of nanomaterials from zero to three-dimensional nanostructures in its discrete and nanocomposites nanotopography on sensing lung cancer biomarkers. Recent researches conducted on definitive nanomaterials and nanocomposites at multiple dimension with distinctive physiochemical property were focused to subside the cases associated with lung cancer through the development of novel biosensors. The hurdles encountered in the recent research and future preference with prognostic clinical lung cancer diagnosis using multidimensional nanomaterials and its composites are presented.

Aluminosilicate Nanocomposite on Genosensor: A Prospective Voltammetry Platform for Epidermal Growth Factor Receptor Mutant Analysis in Non-small Cell Lung Cancer.

Lung cancer is one of the most serious threats to human where 85% of lethal death caused by non-small cell lung cancer (NSCLC) induced by epidermal growth factor receptor (EGFR) mutation. The present research focuses in the development of efficient and effortless EGFR mutant detection strategy through high-performance and sensitive genosensor. The current amplified through 250 µm sized fingers between 100 µm aluminium electrodes indicates the voltammetry signal generated by means of the mutant DNA sequence hybridization. To enhance the DNA immobilization and hybridization, ∼25 nm sized aluminosilicate nanocomposite synthesized from the disposed joss fly ash was deposited on the gaps between aluminium electrodes. The probe, mutant (complementary), and wild (single-base pair mismatch) targets were designed precisely from the genomic sequences denote the detection of EGFR mutation. Fourier-transform Infrared Spectroscopy analysis was performed at every step of surface functionalization evidences the relevant chemical bonding of biomolecules on the genosensor as duplex DNA with peak response at 1150 cm-1 to 1650 cm-1. Genosensor depicts a sensitive EGFR mutation as it is able to detect apparently at 100 aM mutant against 1 µM DNA probe. The insignificant voltammetry signal generated with wild type strand emphasizes the specificity of genosensor in the detection of single base pair mismatch. The inefficiency of genosensor in detecting EGFR mutation in the absence of aluminosilicate nanocomposite implies the insensitivity of genosensing DNA hybridization and accentuates the significance of aluminosilicate. Based on the slope of the calibration curve, the attained sensitivity of aluminosilicate modified genosensor was 3.02E-4 A M-1. The detection limit of genosensor computed based on 3σ calculation, relative to the change of current proportional to the logarithm of mutant concentration is at 100 aM.