Search Results (472)

The α⁰-thalassemia Southeast Asian (--^SEA) and Thai (--^THAI) deletions comprise a significant global health burden, with high prevalence in Southeast Asia. Conventional methods like polymerase chain reaction (PCR) are limited in resource-poor settings due to complexity, time, and cost. This assay was developed as a rapid and simple option for point-of-care (POC) applications. Three assays specific for wild-type DNA, --^SEA and --^THAI deletions utilized recombinase polymerase amplification (RPA) for rapid DNA amplification at a constant low temperature of 37 °C, with a run time of 10 min, eliminating the need for a thermal cycler. The amplified products were detected using the highly specific CRISPR-Cas12a system and read visually via fluorescence or combined with a lateral flow assay (LFA) in 30 min. Successful engineering of a synthetic PAM site enabled Cas12a recognition of the --^SEA target despite the absence of a suitable natural PAM. The platform demonstrated high accuracy in 74 blinded clinical samples, achieving a diagnostic accuracy of 98.20%, a sensitivity of 94.59%, and a specificity of 100% across 222 per-target evaluations. Detection from non-invasive samples achieved 80% to 100% accuracy, depending on the extraction method and readout format used. Our platform showed potential to meet several ASSURED criteria for diagnostic tests in resource-limited settings, demonstrating high potential for widespread use in genetic screening for α⁰-thalassemia (--^SEA) and (--^THAI). Full article

MicroRNAs (miRNAs) represent highly promising biomarkers for non-invasive clinical diagnostics. To overcome the limitations of conventional methods, Field-Effect Transistor biosensors (bioFETs) emerge as compelling alternatives by providing label-free and real-time readouts. However, bioFETs employing canonical DNA/RNA probes face fundamental limitations due to the Debye screening effect. Peptide Nucleic Acid (PNA), a synthetic nucleic acid analog featuring an uncharged peptide-like backbone, offers an effective solution. PNA’s electroneutrality allows for efficient hybridization at a low ionic strength, thereby mitigating Debye screening constraints. PNA also offers additional key advantages, including high specificity and enzymatic resistance. This work examines recent advancements in PNA–bioFETs, highlighting their potential for the development of robust platforms for clinical miRNA evaluation. Surface Plasmon Resonance (SPR) and Atomic Force Spectroscopy (AFS) are valuable tools to preliminarily characterize probe–target interactions. Key approaches, focused on implementing alternative PNA probe designs and optimizing Self-Assembled Monolayer (SAM) organization, are outlined. By presenting a cross-study comparison that correlates each strategy with the analytical outcomes of a custom-designed bioFET platform targeting miRNA 155, complemented by an overview of PNA–bioFETs across other miRNAs, we illustrate how the synergy between PNA and bioFET can approach reliable miRNA detection, paving the way for the development of Point-of-Care (POC) devices. Full article

Early detection of colorectal cancer (CRC) is challenging in low-resource settings because access to colonoscopy and centralized laboratory testing is limited. Urine-based metabolite biomarkers offer a non-invasive alternative for CRC triage, but translating a laboratory assay into a point-of-care (PoC) system requires standardized fluid handling, operator-independent timing, field-compatible reagents, and quantitative calibration. Here, we present a low-cost, semi-automated PoC platform integrating a validated, sequential, three-metabolite CRC-biomarker assay with robotic fluid handling, a motorized chromatographic module, an optical reader, Bluetooth electronics, and tablet-guided control. The platform measures urinary diacetylspermine and hippuric acid, with creatinine serving as a normalization reference, and reports absolute quantitative concentrations. Automated dilution, tube positioning, timed incubation, controlled column elution, and software-guided transfer reduce operator-dependent variation. Using pooled urine samples spiked at clinically relevant concentrations, the creatinine assay showed a strong quadratic response (0–50 mM, R² = 0.998), as did the diacetylspermine assay (0–4 μM, R² = 0.979), while the hippuric acid assay showed a linear response of R² = 0.989. The RGB sensor tracked the concentration-dependent trend observed with a laboratory microplate reader (R² = 0.968–0.999). Reagents reformulated as lyophilized or pre-weighed, vacuum-sealed kits withstood accelerated heat and humidity (45 °C/70% RH) testing and international shipping to the pilot site very well. At approximately USD 490 for the complete instrument (including tablet) and roughly USD 7.65 in consumables per screening test, the platform offers a practical, quantitative, and portable route for decentralized CRC screening. Full article

Sexually transmitted infections (STIs) remain a public health challenge, with rising incidence rates and significant loss to follow-up in traditional testing models. Point-of-care (POC) testing promises same-day diagnosis and treatment, potentially transforming STI management. We conducted a narrative review with perspective elements of commercially available POC and near-POC tests for STIs, identifying tests through regulatory and product-specific sources including the Food and Drug Administration (FDA), World Health Organization (WHO) prequalification list, Australia’s Therapeutic Goods Administration (TGA), China’s National Medical Products Administration (NMPA), publicly available European regulatory and manufacturer sources for CE-marked in vitro diagnostic devices under the EU In Vitro Diagnostic Regulation framework, and the published literature. Several nucleic acid amplification test (NAAT)-based platforms now deliver results rapidly with excellent sensitivity and specificity for common bacterial and protozoal STIs, including Chlamydia trachomatis, Neisseria gonorrhoeae, and Trichomonas vaginalis. For viral STIs, POC testing is most mature for human immunodeficiency virus (HIV), with numerous approved rapid antibody tests, although viral load and resistance testing remain laboratory-based. Most NAAT platforms require dedicated instruments, specialized training, and significant infrastructure. At-home testing options are emerging but face challenges with sensitivity, public health surveillance, and regulatory oversight. In conclusion, POC testing for STIs is advancing but not yet fully realized, and true POC tests (those usable at the bedside without laboratory support) remain limited. While rapid NAAT platforms represent significant progress over traditional laboratory-based testing, barriers to true POC implementation persist. Future developments in antimicrobial resistance detection, multiplex testing, instrument-free platforms, and integration of at-home testing with public health surveillance will be critical to achieving broader STI testing and control. Full article

In this study, native inulin was phosphorylated and used as a stabilizer in duck-fat-based whipped cream to investigate its effects on emulsion stability, rheological properties, whipping performance, and sensory quality. Characterization of phosphorylated inulin showed a phosphorus content of 0.38% and a degree of substitution (DS) of 0.020. FTIR analysis revealed phosphate-associated absorption bands at approximately 1220 and 992 cm⁻¹, attributed to P = O and P–O–C stretching vibrations, respectively, supporting the successful phosphorylation of inulin. Within the PI series, the average particle size decreased as the phosphorylated-inulin addition level increased, reaching 629 ± 5 nm in PI-12, consistent with the CLSM observations. The absolute magnitude of the negative zeta potential generally increased with the phosphorylated-inulin addition level. All emulsions exhibited shear-thinning behavior, with storage modulus (G′) exceeding loss modulus (G″). Higher phosphorylated-inulin addition levels were associated with longer whipping times and lower overrun. No measurable serum loss was detected for PI-6, PI-9, or PI-12. Air bubbles appeared smaller and more uniformly distributed at higher phosphorylated-inulin addition levels. The preliminary sensory assessment showed higher mean texture and smoothness scores for formulations containing 12 g/batch. Within the phosphorylated-inulin series, PI-6 (6 g/batch) was the lowest tested addition level at which no measurable serum loss was detected after 3 h at 22 °C, although higher addition levels further increased viscosity and viscoelastic moduli. Increasing the phosphorylated-inulin addition level above PI-6 further prolonged whipping time and reduced overrun. Further studies should evaluate storage stability and oxidative stability to validate long-term performance. Full article

Point-of-care (POC) technologies are promising tools to decentralize and accelerate the diagnosis of rare diseases. Among them, electrochemical immunosensors offer advantages such as high sensitivity, low cost, portability, low sample consumption, and suitability for use in resource-limited settings. However, the performance of these devices is dependent on electrode surface properties, which influence electron transfer, biomolecule immobilization, and analytical sensitivity. In this work, screen-printed carbon electrodes (SPCEs) were modified through two strategies: (i) gold electrodeposition and (ii) cold plasma treatment. The modified electrodes were functionalized with EDC/NHS, followed by the immobilization of anti-SMN antibodies and electrochemical characterization using cyclic voltammetry and differential pulse voltammetry. The impact of each modification approach on the electrochemical response and reproducibility of the sensor was evaluated. Gold electrodeposition resulted in higher and more reproducible electrochemical responses, demonstrating improved electron transfer properties and surface homogeneity. The primary objective of this study was to investigate how different surface modification strategies affect the electrochemical performance of SPCE-based immunosensors, employing the detection of Survival Motor Neuron (SMN) protein, a biomarker associated with Spinal Muscular Atrophy (SMA), as a proof-of-concept application. The resulting platform successfully differentiated specific and non-specific protein recognition events through distinct electrochemical patterns, demonstrating the suitability of gold-modified SPCEs for immunosensing applications. These findings provide insights into the influence of surface engineering strategies on sensor performance and support the future development of optimized electrochemical platforms for biomarker detection. Full article

With the increasing softwarization of satellite payloads and the evolution of 6G Non-Terrestrial Networks (NTNs), securing the onboard software supply chain has become critical for mission-critical space communications. Traditional static software composition analysis (SCA) often generates excessive false positives in resource-constrained satellite environments. To address this issue, this paper proposes SSCRPDM, a CVE-oriented static–dynamic risk detection method based on bytecode instrumentation. SSCRPDM parses CVE semantics to guide function-level minimal instrumentation, quantifies risk by jointly considering runtime reachability, parameter controllability, and sanitizer effectiveness, and employs deterministic proof-of-concept (PoC) verification as the primary decision mechanism, with a large language model used only for auxiliary interpretation. Specifically, SCA first identifies components containing known vulnerabilities. Runtime instrumentation then traces critical invocation paths and verifies the reachability of vulnerable functions. Finally, CVE-specific PoC payloads are executed under the current defense context to determine actual exploitability. Experimental results show that SSCRPDM effectively filters alerts caused by “zombie components” and significantly improves evaluation accuracy through exploitability verification. By bridging static version matching and dynamic threat validation, SSCRPDM substantially reduces false positives and provides a precise and proactive solution for satellite software supply chain governance. Full article

Clarifying SOC sequestration via physical and microbial processes is key for improving farmland fertility, yet the relative contributions of agronomic practices to carbon fractions and aggregate sizes remain unclear. This study (2010–2019, rice–wheat rotation, Funiu Mountain eastern plain, central China) examined tillage and organic amendment effects on SOC dynamics and underlying mechanisms across aggregate sizes under six treatments (conventional/reduced tillage with no fertilizer, chemical fertilizer, or organic manure). SOC, particulate organic carbon (POC), and mineral-incorporated organic carbon (MOC) were measured in bulk soil and water-stable aggregates (>2000, 250–2000, 53–250, <53 μm), and physical fractionation and phospholipid fatty acid (PLFA) analysis were conducted to assess interactions among aggregates, carbon quality, and microbial communities. Results showed that, compared with conventional tillage without fertilization, both conventional tillage and reduced tillage with organic manure significantly increased bulk SOC by 92–122% and macroaggregate (>250 μm) mass by 15–110%. The combined application of organic manure and reduced tillage redirected SOC from micro- to macroaggregates. Moreover, POC and MOC were the primary contributors to bulk SOC, with POC showing a strong direct effect on SOC accumulation. Furthermore, a positive priming effect was detected exclusively in macroaggregates, identifying them as key sites for SOC turnover and confirming that optimized tillage with manure shifts aggregates to larger sizes and boosts SOC through physical protection. The micro-to-macro cascade offers a robust framework for SOC dynamics, and its persistence under diverse climates warrants future research for sustainable management. Full article

In recent years, nanopore technology has enhanced analyte detection, enabled higher resolution and achieved single-molecule sensing capability. Aptamer-conjugated nanopore sensing technology combines the high specificity of aptamers with the single-molecule resolution of nanopores. By anchoring aptamers to biological, solid-state or hybrid nanopores, target binding events produce distinct electrical signatures that allow sensitive and label-free detection. This approach enables real-time monitoring of small molecules, proteins, and even pathogens, with promising applications in diagnostics, drug screening, environmental monitoring, etc. Hybrid biological/solid state devices produce robust signals and are suitable for PoC applications. The aptamers “magic bullets” have also been exploited to develop single-molecule antigen detection using nanopores, which offers a promising alternative for accurate virus testing to contain their transmission. Chemical conjugation of aptamers to nanopore interfaces improves selectivity for peptides/amino acids and expands robustness for practical samples. Aptamer-based nanopipettes offer high analytical precision by enabling label-free, real-time detection of target molecules in ultra-small sample volumes. This review maps aptamer–nanopore integration across biological, solid-state, and hybrid platforms. It also explores various types of aptamers integrated into nanopore platforms that cater to precise, single-molecule recognition, paving the way for highly sensitive, portable diagnostics and next-generation therapeutic monitoring tools. Full article

Early detection of lung cancer remains challenging due to the extremely low concentrations of disease-specific biomarkers, which limit the development of highly sensitive and reliable point-of-care (PoC) diagnostic devices. Electrochemical biosensors integrating graphene oxide (GO) and gold nanoparticles (AuNPs) have emerged as promising platforms for the rapid, sensitive, and selective detection of lung cancer biomarkers, enabling more timely diagnosis. Biomarkers such as carcinoembryonic antigen (CEA), cytokeratin-19 fragments (CYFRA 21-1), neuron-specific enolase (NSE), and circulating tumour DNA are increasingly investigated for PoC applications since they can be detected in various biological fluids associated with lung cancer. Nanocomposite materials, particularly GO/AuNP hybrids, provide synergistic advantages by combining the large surface area and abundant functional groups of GO for stable immobilization of biorecognition elements with the excellent conductivity and bioconjugation capability of AuNPs that enhance signal transduction. This review critically discusses key biomarker targets for lung cancer, the properties of GO and Au in biosensing, and the role of AuNP/GO nanocomposites in improving biosensor performance. It further examines the application of electrochemical biosensors for lung cancer biomarker detection, highlighting recent developments. Additionally, the review outlines current challenges limiting clinical translation and PoC implementation, provides recommendations to address these barriers, and discusses future perspectives for improving the detection of low-abundance biomarkers for early lung cancer diagnosis. Ultimately, these technologies seem promising for the development of rapid diagnostic tools equivalent to established platforms such as lateral-flow immunoassays. Full article

To address the low efficiency of manual quality grading for Pleurotus ostreatus in factory-scale production and the difficulty existing computer vision models face in balancing high localization accuracy with real-time edge deployment for food processing, a lightweight non-destructive detection model named POC-DETR-Prune is proposed. Based on an improved RTDETR framework, FasterNet is introduced to optimize feature extraction, reducing memory access latency while ensuring deep feature representation for complex food morphologies. A Small Object Enhancement Pyramid (SOEP) module is designed to mitigate the loss of subtle features caused by dense mushroom clustering. Furthermore, the Inner-MPDIoU loss function is proposed to significantly improve bounding box localization accuracy in highly overlapped food sorting scenarios. To adapt to industrial hardware constraints, a Random channel pruning strategy compresses computational overhead. Experimental results demonstrate that POC-DETR-Prune achieves a mAP@0.5:0.95 of 83.7% with a computation load of only 38.2 GFLOPs. Deployment testing on the NVIDIA Jetson Orin Nano Super edge computing platform achieves a real-time detection rate of 30.2 FPS. This emerging technology provides a certain level of visual algorithm support for automated quality grading equipment in the edible fungi industry. Full article

Accurate road-event detection and timely alert message dissemination are essential for the safety of connected and automated vehicles. In many scenarios, alert messages must reach not only nearby vehicles but also remote stakeholders, such as traffic management centers, cloud services, and infrastructure operators. This requirement motivates the adoption of cellular-based communication technologies in addition to short-range vehicle-to-everything (V2X) communications for data dissemination. In this work, we investigate vehicle-to-network-to-everything (V2N2X) communications for the dissemination of alert messages generated after the on-board detection of hazardous road events through machine learning (ML) algorithms. Although V2N2X connectivity is well suited for extending data dissemination beyond the local vehicular environment, its capability to guarantee prompt message delivery under strict latency constraints remains an open challenge, particularly when ML inference is integrated into the end-to-end processing pipeline. To address this issue, we develop and experimentally evaluate a proof-of-concept (PoC) platform that combines real-time road-event detection with relevant message dissemination towards both nearby and remote recipients. The proposed framework leverages 5G connectivity and publish/subscribe messaging protocols. The experimental results showcase that dissemination latency is highly influenced by both the adopted type of 5G deployment (private versus commercial networks) and the load conditions at the message broker. Full article

Nitrofurantoin (NFT) is a widely used antibiotic that requires sensitive and reliable detection due to its potential environmental and health impacts. In this study, a poly(O-cresolphthalein complexone)/electrochemically reduced graphene oxide (POC/ERGO) nanocomposite was electrochemically fabricated via one-step process and applied to a modified GCE for the electrochemical detection of nitrofurantoin. The sensing performance of the POC/ERGO-GCE was evaluated using CV and DPV. The developed sensor exhibited a wide linear detection range from 1 to 500 μM, and a low detection limit of 78.90 nM as determined by DPV. In addition, it demonstrated excellent anti-interference capability, good reproducibility and selectivity toward NFT, confirming the reliability of the proposed sensing platform. The enhanced performance is attributed to the increased electrochemically active surface area and improved electron transfer properties of the POC/ERGO-GCE. These results indicate that the proposed platform provides a reliable approach for the electrochemical detection of nitrofurantoin and offers a promising foundation for the development of antibiotic sensing systems. Full article

Background/Objectives: Multiplex and point-of-care (POC) diagnostics require each probe to detect one intended target while rejecting many closely related sequences under shared room-temperature conditions. The conventional focus on mismatch count is incomplete: two alignments with the same number of matches and mismatches can have very different off-target risks depending on whether mismatches are clustered or distributed. We introduce a simple visual heuristic that scores mismatch placement rather than mismatch count alone. Methods: Effective complementary island (ECI) score retained matched continuity after subtracting one base for each mismatch- or gap-exposed edge. The score is S_ECI = Σ_i ECI_i^2, and the design margin is ΔS_ECI = S_ECI (intended) − S_ECI (highest-scoring non-intended alignment by ECI). ECI is not a thermodynamic model; thermodynamics (ΔG37) is used separately to verify an adequate sensitivity floor. We retrospectively applied ECI to a fixed 39-target HPV capture-probe benchmark and to a public Affymetrix dataset contrasting clustered versus distributed mismatches at identical or near-identical mismatch counts. Results: In the HPV benchmark, ECI separated intended from off-target in 32/39 panels; ΔG37 favored the intended duplex in 31/39 panels; both layers were concordant in 36/39 panels. In the Affymetrix dataset (n = 8 probes, 2–4 mismatches), S_ECI correlated with reported log2 hybridization intensity (Pearson r = 0.92, p = 0.0014). Within the strict three-mismatch subset (n = 5), S_ECI remained correlated with intensity (r = 0.96; p = 0.010), while ΔG37 was uncorrelated (r = −0.04; p = 0.95), supporting the narrower claim that mismatch placement can affect signal even when mismatch count is fixed. Conclusions: ECI is not a replacement for thermodynamics, BLAST, target-accessibility analysis, empirical optimization, or machine-learning prediction. It adds one actionable readout: where to shift, shorten, or place a limited intentional mismatch so that intended retained continuity stays above the assay floor while the highest-scoring off-target island by ECI is fragmented. We provide a bench-ready workflow for multiplex, room-temperature, and POC probe design. Full article

Bovine Viral Diarrhoea (BVD) is an infectious disease caused by the Bovine Viral Diarrhoea Virus (BVDV), a member of the genus Orthopestivirus. The disease remains endemic across Australian beef and dairy production systems, imposing a multi-million-dollar annual burden on animal health, welfare, and industry sustainability. BVDV can be transmitted both horizontally and vertically, with persistently infected (PI) animals serving as the primary source of infection. Rapid identification and subsequent culling of PI animals are fundamental requirements for any successful eradication program. Currently, Australia’s decentralised, non-compulsory approach places the responsibility of biosecurity on individual producers, resulting in a fragmented national landscape. This review proposes that the strategic deployment of rapid, field-deployable point-of-care (POC) diagnostics serves as the transformative catalyst needed for a coordinated national eradication pathway. POC approaches utilising technologies such as lateral flow assays, nucleic acid amplification tests, and biosensors enable real-time, crush-side diagnosis and high-throughput surveillance, proving effective for early detection and control of infectious diseases. When integrated with robust biosecurity measures and optimised vaccination strategies, these POC advancements offer a scientifically sound and commercially viable pathway toward the systematic eradication of BVDV in the Australian cattle industry. Full article