Development of DNA-Based Lateral Flow Assay for Detection of LDLR Gene Mutation for Familial Hypercholesterolemia.

Background: The techniques for detecting single nucleotide polymorphisms (SNP) require lengthy and complex experimental procedures and expensive instruments that may only be available in some laboratories. Thus, a deoxyribonucleic acid (DNA)-based lateral flow assay (LFA) was developed as a point-of-care test (POCT) diagnostic tool for genotyping. In this study, single nucleotide variation (E101K) in the low-density lipoprotein receptor (LDLR) gene leading to familial hypercholesterolemia (FH) was chosen as a model. Methods: Hypercholesterolemic individuals (n = 103) were selected from the Malaysian Cohort project (UKM Medical Molecular Biology Institute) while the control samples were selected from the Biobank (UKM Medical Molecular Biology Institute). The DNA samples were isolated from whole blood. Polymerase chain reaction (PCR) amplification process was performed using bifunctional labelled primers specifically designed to correspond to the variant that differentiates wild-type and mutant DNA for visual detection on LFA. The variant was confirmed using Sanger sequencing, and the sensitivity and specificity of the LFA detection method were validated using the Agena MassARRAY® technique. Results: Out of 103 hypercholesterolemic individuals, 5 individuals (4.8%) tested positive for E101K, LDLR mutation and the rest, including healthy control individuals, tested negative. This result was concordant with Sanger sequencing and Agena MassARRAY®. These five individuals could be classified as Definite FH, as the DNA diagnosis was confirmed. The sensitivity and specificity of the variant detection by LFA is 100% compared to results using the genotyping method using Agena MassARRAY®. Conclusion: The developed LFA can potentially be used in the POC setting for detecting the E101K variant in the LDLR gene. This LFA can also be used to screen family members with E101K variant in the LDLR gene and is applicable for other SNP's detection.

Formation of Ultimate Thin 2D Crystal of Pt in the Presence of Hexamethylenetetramine.

Platinum naturally crystalizes into a three-dimensional crystal due to its highly symmetrical fcc lattice, with a metallic bond which is non-directional and highly isotropic. This inherently means ultimately that 2D crystals of a few atoms thick growth are hardly available in this material. Here, we discovered that a combinative effect of formic acid reductant and hexamethylenetetramine surfactant during the reduction of their metal ions precursor can realize an ultimate thin 2D crystal growth in platinum. High-resolution transmission electron microscopy and filed-emission electron microscopy analysis have also discovered that the 2D crystal of Pt has 111 facets with a lateral dimension that can be up to more than 5 µm × 2 µm. The thickness of the 2D crystal of Pt is 1.55 nm. A mechanism for obtaining ultimate thin 2D crystal of Pt using the present approach is proposed.

Synthesis of metallic mixed 3R and 2H Nb1+xS2 nanoflakes by chemical vapor deposition.

In this work, we report the synthesis and characterization of mixed phase Nb1+xS2 nanoflakes prepared by chemical vapor deposition. The as-grown samples show a high density of flakes (thickness ∼50 nm) that form a continuous film. Raman and X-ray diffraction data show that the samples consist of both 2H and 3R phases, with the 2H phase containing a high concentration of Nb interstitials. These Nb interstitials sit in between the NbS2 layers to form Nb1+xS2. Cross-sectional Energy Dispersive Spectroscopy analysis with transmission electron microscopy suggests that the 2H Nb1+xS2 region is found in thinner flakes, while 3R NbS2 is observed in thicker regions of the films. The evolution of the phase from 2H Nb1+xS2 to 3R NbS2 may be attributed to the change of the growth environment from Nb-rich at the start of the growth to sulfur-rich at the latter stage. It was also found that the incorporation of Nb interstitials is highly dependent on the temperature of the NbCl5 precursor and the position of the substrate in the furnace. Samples grown at high NbCl5 temperature and with substrate located closer to the NbCl5 source show higher incorporation of Nb interstitials. Electrical measurements show linear I-V characteristics, indicating the metallic nature of the Nb1+xS2 film with relatively low resistivity of 4.1 × 10-3Ω cm.

Infection-Mediated Clinical Biomarkers for a COVID-19 Electrical Biosensing Platform.

The race towards the development of user-friendly, portable, fast-detection, and low-cost devices for healthcare systems has become the focus of effective screening efforts since the pandemic attack in December 2019, which is known as the coronavirus disease 2019 (COVID-19) pandemic. Currently existing techniques such as RT-PCR, antigen-antibody-based detection, and CT scans are prompt solutions for diagnosing infected patients. However, the limitations of currently available indicators have enticed researchers to search for adjunct or additional solutions for COVID-19 diagnosis. Meanwhile, identifying biomarkers or indicators is necessary for understanding the severity of the disease and aids in developing efficient drugs and vaccines. Therefore, clinical studies on infected patients revealed that infection-mediated clinical biomarkers, especially pro-inflammatory cytokines and acute phase proteins, are highly associated with COVID-19. These biomarkers are undermined or overlooked in the context of diagnosis and prognosis evaluation of infected patients. Hence, this review discusses the potential implementation of these biomarkers for COVID-19 electrical biosensing platforms. The secretion range for each biomarker is reviewed based on clinical studies. Currently available electrical biosensors comprising electrochemical and electronic biosensors associated with these biomarkers are discussed, and insights into the use of infection-mediated clinical biomarkers as prognostic and adjunct diagnostic indicators in developing an electrical-based COVID-19 biosensor are provided.

Integration of a Dielectrophoretic Tapered Aluminum Microelectrode Array with a Flow Focusing Technique.

We present the integration of a flow focusing microfluidic device in a dielectrophoretic application that based on a tapered aluminum microelectrode array (TAMA). The characterization and optimization method of microfluidic geometry performs the hydrodynamic flow focusing on the channel. The sample fluids are hydrodynamically focused into the region of interest (ROI) where the dielectrophoresis force (FDEP) is dominant. The device geometry is designed using 3D CAD software and fabricated using the micro-milling process combined with soft lithography using PDMS. The flow simulation is achieved using COMSOL Multiphysics 5.5 to study the effect of the flow rate ratio between the sample fluids (Q1) and the sheath fluids (Q2) toward the width of flow focusing. Five different flow rate ratios (Q1/Q2) are recorded in this experiment, which are 0.2, 0.4, 0.6, 0.8 and 1.0. The width of flow focusing is increased linearly with the flow rate ratio (Q1/Q2) for both the simulation and the experiment. At the highest flow rate ratio (Q1/Q2 = 1), the width of flow focusing is obtained at 638.66 µm and at the lowest flow rate ratio (Q1/Q2 = 0.2), the width of flow focusing is obtained at 226.03 µm. As a result, the flow focusing effect is able to reduce the dispersion of the particles in the microelectrode from 2000 µm to 226.03 µm toward the ROI. The significance of flow focusing on the separation of particles is studied using 10 and 1 µm polystyrene beads by applying a non-uniform electrical field to the TAMA at 10 VPP, 150 kHz. Ultimately, we are able to manipulate the trajectories of two different types of particles in the channel. For further validation, the focusing of 3.2 µm polystyrene beads within the dominant FDEP results in an enhanced manipulation efficiency from 20% to 80% in the ROI.

Titanium dioxide-mediated resistive nanobiosensor for E. coli O157:H7.

A titanium dioxide nanoparticle (TiO2 NP)-mediated resistive biosensor is described for the determination of DNA fragments of Escherichia coli O157:H7 (E. coli O157:H7). The sol-gel method was used to synthesize the TiO2 NP, and microlithography was applied to fabricate the interdigitated sensor electrodes. Conventional E. coli DNA detections are facing difficulties in long-preparation-and-detection-time (more than 3 days). Hence, electronic biosensor was introduced by measuring the current-voltage (I-V) DNA probe without amplification of DNA fragments. The detection scheme is based on the interaction between the electron flow on the sensor and the introduction of negative charges from DNA probe and target DNA. The biosensor has a sensitivity of 1.67 × 1013 Ω/M and a wide analytical range. The limit detection is down to 1 × 10-11 M of DNA. The sensor possesses outstanding repeatability and reproducibility and is cabable to detect DNA within 15 min in a minute-volume sample (1 µL). Graphical abstract Fig. (a) Graphical illustration of electronic biosensor set up and (b) relationship between limit of detection (LOD) and the unaffected poultry samples on E. coli O157:H7.

Ultrasensitive and Highly Selective Graphene-Based Field-Effect Transistor Biosensor for Anti-Diuretic Hormone Detection.

Nephrogenic diabetes insipidus (NDI), which can be congenital or acquired, results from the failure of the kidney to respond to the anti-diuretic hormone (ADH). This will lead to excessive water loss from the body in the form of urine. The kidney, therefore, has a crucial role in maintaining water balance and it is vital to restore this function in an artificial kidney. Herein, an ultrasensitive and highly selective aptameric graphene-based field-effect transistor (GFET) sensor for ADH detection was developed by directly immobilizing ADH-specific aptamer on a surface-modified suspended graphene channel. This direct immobilization of aptamer on the graphene surface is an attempt to mimic the functionality of collecting tube V 2 receptors in the ADH biosensor. This aptamer was then used as a probe to capture ADH peptide at the sensing area which leads to changes in the concentration of charge carriers in the graphene channel. The biosensor shows a significant increment in the relative change of current ratio from 5.76 to 22.60 with the increase of ADH concentration ranging from 10 ag/mL to 1 pg/mL. The ADH biosensor thus exhibits a sensitivity of 50.00 µA· ( g / mL ) - 1 with a limit of detection as low as 3.55 ag/mL. In specificity analysis, the ADH biosensor demonstrated a higher current value which is 338.64 µA for ADH-spiked in phosphate-buffered saline (PBS) and 557.89 µA for ADH-spiked in human serum in comparison with other biomolecules tested. This experimental evidence shows that the ADH biosensor is ultrasensitive and highly selective towards ADH in PBS buffer and ADH-spiked in human serum.

Curvature contribution on anodic aluminium oxide pore formation: a detailed analytical study.

Anodic aluminium oxide (AAO) is a self-organised nanopore that has been widely studied due to the ease of its synthesization and pore properties manipulation. However, pore growth behaviour under different geometrical surfaces is rarely studied, particularly on the effect of combined curved surfaces towards pore growth properties, which is crucial in designing unique porous platform for specific applications. This paper reports study on the decisive effect of curvature surfaces on development of pore structure and properties at a constant potential. In this work, AAO grown on treated convex and concave surfaces were analysed in terms of pore quantity, pore diameter, interpore distance, pore length and other parameters of pore bottom geometry in conjugation with observation of pore cessation, bifurcation, bending and tapering. The unique formation of tapered pore was observed and described. Major factors deciding pore properties under curved surfaces were identified and discussed. We introduced a new parameter for surface quantification known as central inscribed angle, which was identified to be the central factor which decides pore growth behaviour under a curvature. Here, we observed a different trend in growth rate of pores under different curvatures, which oppose the commonly accepted convex > planar > concave pattern. Levelling height was later identified to be the decisive factor in determining growth rate of pores under a curvature at different geometrical location. These findings open up possibility to precisely control and tailor the growing path and pore structures of AAO simply via anodising an Al sheet under combined curvature surfaces, which could be beneficial for future novel applications.

A tapered aluminium microelectrode array for improvement of dielectrophoresis-based particle manipulation.

In this work, the dielectrophoretic force (F(DEP)) response of Aluminium Microelectrode Arrays with tapered profile is investigated through experimental measurements and numerical simulations. A standard CMOS processing technique with a step for the formation of a tapered profile resist is implemented in the fabrication of Tapered Aluminium Microelectrode Arrays (TAMA). The F(DEP) is investigated through analysis of the Clausius-Mossotti factor (CMF) and cross-over frequency (f(xo)). The performance of TAMA with various side wall angles is compared to that of microelectrodes with a straight cut sidewall profile over a wide range of frequencies through FEM numerical simulations. Additionally, electric field measurement (EFM) is performed through scanning probe microscopy (SPM) in order to obtain the region of force focus in both platforms. Results showed that the tapered profile microelectrodes with angles between 60° and 70° produce the highest electric field gradient on the particles. Also, the region of the strongest electric field in TAMA is located at the bottom and top edge of microelectrode while the strongest electric field in microelectrodes with straight cut profile is found at the top corner of the microelectrode. The latter property of microelectrodes improves the probability of capturing/repelling the particles at the microelectrode's side wall.

Synthesis and Characterization of SiO2-Based Graphene Nanoballs Using Copper-Vapor-Assisted APCVD for Thermoelectric Application.

This study describes a method by which to synthesize SiO2-based graphene nanoballs (SGB) using atmospheric pressure chemical vapor deposition (APCVD) with copper vapor assistance. This method should solve the contamination, damage, and high costs associated with silica-based indirect graphene synthesis. The SGB was synthesized using APCVD, which was optimized using the Taguchi method. Multiple synthesis factors were optimized and investigated to find the ideal synthesis condition to grow SGB for thermoelectric (TE) applications. Raman spectra and FESEM-EDX reveal that the graphene formed on the silicon nanoparticles (SNP) is free from copper. The prepared SGB has excellent electrical conductivity (75.0 S/cm), which shows better results than the previous report. Furthermore, the SGB nanofillers in bismuth telluride (Bi2Te3) nanocomposites as TE materials exhibit a significant increment in Seebeck coefficients (S) compared to the pure Bi2Te3 sample from 109 to 170 µV/K at 400 K, as well as electrical resistivity decrement. This approach would offer a simple strategy to improve the TE performance of commercially available TE materials, which is critical for large-scale industrial applications.

Air-Processable Perovskite Solar Cells by Hexamine Molecule Phase Stabilization.

Perovskite solar cells have emerged as a potential energy alternative due to their low cost of fabrication and high power conversion efficiency. Unfortunately, their poor ambient stability has critically limited their industrialization and application in real environmental conditions. Here, we show that by introducing hexamine molecules into the perovskite lattice, we can enhance the photoactive phase stability, enabling high-performance and air-processable perovskite solar cells. The unencapsulated and freshly prepared perovskite solar cells produce a power conversion efficiency of 16.83% under a 100 mW cm-2 1.5G solar light simulator and demonstrate high stability properties when being stored for more than 1500 h in humid air with relative humidity ranging from 65 to 90%. We envisage that our findings may revolutionize perovskite solar cell research, pushing the performance and stability to the limit and bringing the perovskite solar cells toward industrialization.

Geometrical Characterisation of TiO2-rGO Field-Effect Transistor as a Platform for Biosensing Applications.

The performance of the graphene-based field-effect transistor (FET) as a biosensor is based on the output drain current (Id). In this work, the signal-to-noise ratio (SNR) was investigated to obtain a high-performance device that produces a higher Id value. Using the finite element method, a novel top-gate FET was developed in a three-dimensional (3D) simulation model with the titanium dioxide-reduced graphene oxide (TiO2-rGO) nanocomposite as the transducer material, which acts as a platform for biosensing application. Using the Taguchi mixed-level method in Minitab software (Version 16.1.1), eighteen 3D models were designed based on an orthogonal array L18 (6134), with five factors, and three and six levels. The parameters considered were the channel length, electrode length, electrode width, electrode thickness and electrode type. The device was fabricated using the conventional photolithography patterning technique and the metal lift-off method. The material was synthesised using the modified sol-gel method and spin-coated on top of the device. According to the results of the ANOVA, the channel length contributed the most, with 63.11%, indicating that it was the most significant factor in producing a higher Id value. The optimum condition for the highest Id value was at a channel length of 3 µm and an electrode size of 3 µm × 20 µm, with a thickness of 50 nm for the Ag electrode. The electrical measurement in both the simulation and experiment under optimal conditions showed a similar trend, and the difference between the curves was calculated to be 28.7%. Raman analyses were performed to validate the quality of TiO2-rGO.

Detection of SARS-CoV-2 in Environment: Current Surveillance and Effective Data Management of COVID-19.

Since diagnostic laboratories handle large COVID-19 samples, researchers have established laboratory-based assays and developed biosensor prototypes. Both share the same purpose; to ascertain the occurrence of air and surface contaminations by the SARS-CoV-2 virus. However, the biosensors further utilize internet-of-things (IoT) technology to monitor COVID-19 virus contamination, specifically in the diagnostic laboratory setting. The IoT-capable biosensors have great potential to monitor for possible virus contamination. Numerous studies have been done on COVID-19 virus air and surface contamination in the hospital setting. Through reviews, there are abundant reports on the viral transmission of SARS-CoV-2 through droplet infections, person-to-person close contact and fecal-oral transmission. However, studies on environmental conditions need to be better reported. Therefore, this review covers the detection of SARS-CoV-2 in airborne and wastewater samples using biosensors with comprehensive studies in methods and techniques of sampling and sensing (2020 until 2023). Furthermore, the review exposes sensing cases in public health settings. Then, the integration of data management together with biosensors is well explained. Last, the review ended with challenges to having a practical COVID-19 biosensor applied for environmental surveillance samples.

State-of-the-Art on Functional Titanium Dioxide-Integrated Nano-Hybrids in Electrical Biosensors.

Biosensors operating based on electrical methods are being accelerated toward rapid and efficient detection that improve the performance of the device. Continuous study in nano- and material-sciences has led to the inflection with properties of nanomaterials that fit the trend parallel to the biosensor evolution. Advancements in technology that focuses on nano-hybrid are being used to develop biosensors with better detection strategies. In this sense, titanium dioxide (TiO2) nanomaterials have attracted extensive interest in the construction of electrical biosensors. The formation of TiO2 nano-hybrid as an electrical transducing material has revealed good results with high performance. The modification of the sensing portion with a combination (nano-hybrid form) of nanomaterials has produced excellent sensors in terms of stability, reproducibility, and enhanced sensitivity. This review highlights recent research advancements with functional TiO2 nano-hybrid materials, and their victorious story in the construction of electrical biosensors are discussed. Future research directions with commercialization of these devices and their extensive utilizations are also discussed.

Lifting the Veil: Characteristics, Clinical Significance, and Application of ß-2-Microglobulin as Biomarkers and Its Detection with Biosensors.

Because ß-2-microglobulin (ß2M) is a surface protein that is present on most nucleated cells, it plays a key role in the human immune system and the kidney glomeruli to regulate homeostasis. The primary clinical significance of ß2M is in dialysis-related amyloidosis, a complication of end-stage renal disease caused by a gradual accumulation of ß2M in the blood. Therefore, the function of ß2M in kidney-related diseases has been extensively studied to evaluate its glomerular and tubular functions. Because increased ß2M shedding due to rapid cell turnover may indicate other underlying medical conditions, the possibility to use ß2M as a versatile biomarker rose in prominence across multiple disciplines for various applications. Therefore, this work has reviewed the recent use of ß2M to detect various diseases and its progress as a biomarker. While the use of state-of-the-art ß2M detection requires sophisticated tools, high maintenance, and labor cost, this work also has reported the use of biosensor to quantify ß2M over the past decade. It is hoped that a portable and highly efficient ß2M biosensor device will soon be incorporated in point-of-care testing to provide safe, rapid, and reliable test results.

Protein Albumin Manipulation and Electrical Quantification of Molecular Dielectrophoresis Responses for Biomedical Applications.

Research relating to dielectrophoresis (DEP) has been progressing rapidly through time as it is a strong and controllable technique for manipulation, separation, preconcentration, and partitioning of protein. Extensive studies have been carried out on protein DEP, especially on Bovine Serum Albumin (BSA). However, these studies involve the usage of dye and fluorescent probes to observe DEP responses as the physical properties of protein albumin molecular structure are translucent. The use of dye and the fluorescent probe could later affect the protein's physiology. In this article, we review three methods of electrical quantification of DEP responses: electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV) and capacitance measurement for protein BSA DEP manipulation. The correlation of these methods with DEP responses is further discussed. Based on the observations on capacitance measurement, it can be deduced that the electrical quantifying method is reliable for identifying DEP responses. Further, the possibility of manipulating the protein and electrically quantifying DEP responses while retaining the original physiology of the protein and without the usage of dye or fluorescent probe is discussed.

Transdermal Maltose-Based Microneedle Patch as Adjunct to Enhance Topical Anesthetic before Intravenous Cannulation of Pediatric Thalassemic Patients Receiving Blood Transfusion: A Randomized Controlled Trial Protocol.

Intravenous cannulation is experientially traumatic to children. To minimize this, EMLA® is applied on the would-be-cannulated area before IV cannula insertion. However, the time to achieve its maximum efficacy may be affected due to incomplete cutaneous absorption and the duration of application. The latter may be a limiting factor in a busy healthcare facility. The usage of dissolvable maltose microneedles may circumvent this problem by introducing micropores that will facilitate EMLA® absorption. A randomized phase II cross-over trial will be conducted to compare the Visual Analogue Scale (VAS) pain scores and skin conductance algesimeter index between 4 different interventions (1 fingertip unit (FTU) of EMLA® with microneedle patch for 30 min before cannulation; 0.5 FTU of EMLA® with microneedle patch for 30 min; 1 FTU of EMLA® with microneedle for 15 min; 1 FTU of EMLA® with sham patch for 30 min). A total of 26 pediatric patients with thalassemia aged between 6 and 18 years old and requiring blood transfusion will be recruited in this trial. During the visits, the VAS scores and skin conductance algesimeter index at venous cannulation will be obtained using the VAS rulers and PainMonitor™ machine, respectively. The trial will commence in August 2021 and is anticipated to end by August 2022.

Potentials of MicroRNA in Early Detection of Ovarian Cancer by Analytical Electrical Biosensors.

The importance of nanotechnology in medical applications especially with biomedical sensing devices is undoubted. Several medical diagnostics have been developed by taking the advantage of nanomaterials, especially with electrical biosensors. Biosensors have been predominantly used for the quantification of different clinical biomarkers toward detection, screening, and follow-up the treatment. At present, ovarian cancer is one of the severe complications that cannot be identified until it becomes most dangerous as the advanced stage. Based on the American Cancer Society, 20% of cases involved in the detection of ovarian cancer are diagnosed at an early stage and 80% diagnosed at the later stages. The patient just has a common digestive problem and stomach ache as early symptoms and people used to ignore these symptoms. Micro ribonucleic acid (miRNA) is classified as small non-coding RNAs, their expressions change due to the association of cancer development and progression. This article reviews and discusses on the currently available strategies for the early detection of ovarian cancers using miRNA as a biomarker associated with electrical biosensors. A unique miRNA-based biomarker detections are specially highlighted with biosensor platforms to diagnose ovarian cancer.

Review of Thermoelectric Generators at Low Operating Temperatures: Working Principles and Materials.

Thermoelectric generators (TEGs) are a form of energy harvester and eco-friendly power generation system that directly transform thermal energy into electrical energy. The thermoelectric (TE) method of energy harvesting takes advantage of the Seebeck effect, which offers a simple solution for fulfilling the power-supply demand in almost every electronics system. A high-temperature condition is commonly essential in the working mechanism of the TE device, which unfortunately limits the potential implementation of the device. This paper presents an in-depth analysis of TEGs at low operating temperature. The review starts with an extensive description of their fundamental working principles, structure, physical properties, and the figure of merit (ZT). An overview of the associated key challenges in optimising ZT value according to the physical properties is discussed, including the state of the art of the advanced approaches in ZT optimisation. Finally, this manuscript summarises the research status of Bi2Te3-based semiconductors and other compound materials as potential materials for TE generators working at low operating temperatures. The improved TE materials suggest that TE power-generation technology is essential for sustainable power generation at near-room temperature to satisfy the requirement for reliable energy supplies in low-power electrical/electronics systems.

Mechanical Energy Sensing and Harvesting in Micromachined Polymer-Based Piezoelectric Transducers for Fully Implanted Hearing Systems: A Review.

The paper presents a comprehensive review of mechanical energy harvesters and microphone sensors for totally implanted hearing systems. The studies on hearing mechanisms, hearing losses and hearing solutions are first introduced to bring to light the necessity of creating and integrating the in vivo energy harvester and implantable microphone into a single chip. The in vivo energy harvester can continuously harness energy from the biomechanical motion of the internal organs. The implantable microphone executes mechanoelectrical transduction, and an array of such structures can filter sound frequency directly without an analogue-to-digital converter. The revision of the available transduction mechanisms, device configuration structures and piezoelectric material characteristics reveals the advantage of adopting the polymer-based piezoelectric transducers. A dual function of sensing the sound signal and simultaneously harvesting vibration energy to power up its system can be attained from a single transducer. Advanced process technology incorporates polymers into piezoelectric materials, initiating the invention of a self-powered and flexible transducer that is compatible with the human body, magnetic resonance imaging system (MRI) and the standard complementary metal-oxide-semiconductor (CMOS) processes. The polymer-based piezoelectric is a promising material that satisfies many of the requirements for obtaining high performance implantable microphones and in vivo piezoelectric energy harvesters.