Search Results (368)

Background: Influenza viruses are major pathogens responsible for respiratory infections and pose significant risks to densely populated urban areas. RT-qPCR has made substantial contributions in controlling virus transmission during previous COVID-19 epidemics, but it faces challenges in terms of detection time for large sample sizes and susceptibility to nucleic acid contamination. Methods: Our study designed loop-mediated isothermal amplification primers for three common influenza viruses: A/H3N2, A/H1N1, and B/Victoria, and utilized a 4-channel microfluidic chip to achieve simultaneous detection. The chip initiates amplification by centrifugation and allows testing of up to eight samples at a time. Results: By creating a closed amplification system in the microfluidic chip, aerosol-induced nucleic acid contamination can be prevented through physically isolating the reaction from the operating environment. The chip can specifically detect A/H1N1, A/H3N2, and B/Victoria and has no signal for other common respiratory viruses. The testing process can be completed within 1 h and can be sensitive to viral RNA at concentrations as low as 10⁻³ ng/μL for A/H1N1 and A/H3N2 and 10⁻¹ ng/μL for B/Victori. A total of 296 virus swab samples were further analyzed using the microfluidic chip method and compared with the classical qPCR method, which resulted in high consistency. Conclusions: Our chip enables faster detection of influenza virus and avoids nucleic acid contamination, which is beneficial for POCT establishment and has lower requirements for the operating environment. Full article

Background/Objectives: Direct oral anticoagulants (DOACs) are now the guideline-recommended alternative to vitamin K antagonists (VKAs) for long-term anticoagulation in patients with non-valvular atrial fibrillation. However, accurately assessing their impact on ischemic stroke outcomes remains challenging, primarily due to uncertainty regarding anticoagulation status at the time of hospital admission. This preliminary study addresses this gap by using point-of-care testing (POCT) to confirm DOAC activity at bedside, allowing for a more accurate comparison of 90-day functional outcomes between anticoagulated and non-anticoagulated stroke patients. Methods: We conducted a retrospective cohort study of 786 ischemic stroke patients admitted to the University of Pécs between February 2023 and February 2025. Active DOAC therapy was confirmed using the ClotPro^® viscoelastic testing platform, with ecarin Clotting Time (ECT) employed for thrombin inhibitors and Russell’s Viper Venom (RVV) assays for factor Xa inhibitors. Patients were categorized as non-anticoagulated (n = 767) or DOAC-treated with confirmed activity (n = 19). Mahalanobis distance-based matching was applied to account for confounding variables including age, sex, pre-stroke modified Rankin Scale (mRS), and National Institutes of Health Stroke Scale (NIHSS) scores at admission and 72 h post-stroke. The primary outcome was the change in mRS from baseline to 90 days. Statistical analysis included ordinary least squares (OLS) regression and principal component analysis (PCA). Results: After matching, 90-day functional outcomes were comparable between groups (mean mRS-shift: 2.00 in DOAC-treated vs. 1.78 in non-anticoagulated; p = 0.745). OLS regression showed no significant association between DOAC status and recovery (p = 0.599). In contrast, NIHSS score at 72 h (p = 0.004) and age (p = 0.015) were significant predictors of outcome. PCA supported these findings, identifying stroke severity as the primary driver of outcome. Conclusions: This preliminary analysis suggests that ischemic stroke patients with confirmed active DOAC therapy at admission may achieve 90-day functional outcomes comparable to those of non-anticoagulated patients. The integration of bedside POCT enhances the reliability of anticoagulation assessment and underscores its clinical value for real-time management in acute stroke care. Larger prospective studies are needed to validate these findings and to further refine treatment strategies. Full article

Since the initial use of biological ion channels to detect single-stranded genomic base pair differences, label-free and highly sensitive resistive pulse sensing (RPS) with nanopores has made remarkable progress in single-molecule analysis. By monitoring transient ionic current disruptions caused by molecules translocating through a nanopore, this technology offers detailed insights into the structure, charge, and dynamics of the analytes. In this work, the RPS platforms based on biological, solid-state, and other sensing pores, detailing their latest research progress and applications, are reviewed. Their core capability is the high-precision characterization of tiny particles, ions, and nucleotides, which are widely used in biomedicine, clinical diagnosis, and environmental monitoring. However, current RPS methods involve bottlenecks, including limited sensitivity (weak signals from sub-nanometer targets with low SNR), complex sample interference (high false positives from ionic strength, etc.), and field consistency (solid-state channel drift, short-lived bio-pores failing POCT needs). To overcome this, bio-solid-state fusion channels, in-well reactors, deep learning models, and transfer learning provide various options. Evolving into an intelligent sensing ecosystem, RPS is expected to become a universal platform linking basic research, precision medicine, and on-site rapid detection. Full article

Vitamin D is the only vitamin that is conditionally essential, as it is synthesized from precursors after UV light exposure, whilst also being obtained from the diet. It has numerous health benefits, with deficiency becoming a major concern globally, such that dietary supplementation has more recently achieved vital importance to maintain satisfactory levels. In recent years, measurements made from blood have, therefore, become critical to determine the status of vitamin D levels in individuals and the larger population. Tests for vitamin D have routinely relied on laboratory analysis with sophisticated equipment, often being slow and costly, whilst rapid immunoassays have suffered from poor specificity and sensitivity. Here, we have evaluated a new rapid immunoassay test on the market (Rapi-D & IgLoo) to quickly and accurately measure vitamin D levels in small capillary blood specimens and compared this to measurements made using the standard laboratory method of liquid chromatography and mass spectrometry. Our results show that vitamin D can be measured very quickly and over a broad range using the new method, as well as correlate relatively well with standard laboratory testing; however, it cannot be fully relied upon currently to accurately diagnose deficiency or sufficiency in individuals. Our statistical and comparative analyses find that the rapid immunoassay with digital quantification significantly overestimates vitamin D levels, leading to diminished diagnosis of vitamin D deficiency. The speed and simplicity of the rapid method will likely provide advantages in various healthcare settings; however, further calibration of this rapid method and testing parameters for improving quantification of vitamin D from capillary blood specimens is required before integration of it into clinical decision-making pathways. Full article

Patient blood management (PBM) is a multidisciplinary approach aimed at improving patient outcomes through targeted anemia treatment that minimizes allogeneic blood transfusions, employs blood conservation techniques, and avoids inappropriate use of blood product transfusions. Viscoelastic testing (VET) techniques, such as thromboelastography (TEG) and rotational thromboelastometry (ROTEM), have led to significant advancements in PBM. These techniques offer real-time whole-blood assessment of hemostatic function. This provides the clinician with a more complete hemostasis perspective compared to that provided by conventional coagulation tests (CCTs), such as the prothrombin time (PT) and the activated partial thromboplastin time (aPTT), which only assess plasma-based coagulation. VET does this by mapping the complex processes of clot formation, stability, and breakdown (i.e., fibrinolysis). As a result of real-time whole-blood coagulation assessment during hemorrhage, hemostasis can be achieved through targeted transfusion therapy. This approach helps fulfill an objective of PBM by helping to reduce unnecessary transfusions. However, challenges remain that limit broader adoption of VET, particularly in hospital settings. Of these, standardization and the high cost of the devices are those that are faced the most. This discussion highlights the potential of VET application in PBM to guide blood-clotting therapies and improve outcomes in patients with coagulopathies from various causes that result in hemorrhage. Another aim of this discussion is to highlight the limitations of implementing these technologies so that appropriate measures can be taken toward their wider integration into clinical use. Full article

Background/Objectives: Norovirus is a major cause of acute gastroenteritis worldwide. Considering its highly infectious and transmissible nature, rapid and accurate diagnostic tools are of utmost importance for the effective control of outbreaks in the context of point-of-care testing (POCT). In this study, we developed a genogroup-specific multiplex reverse transcriptase loop-mediated isothermal amplification assay to detect the human norovirus genogroups I and II (GI and GII, respectively). Methods: For the comprehensive detection of clinically relevant genotypes, two sets of primers were incorporated into the assays targeting the RdRp-VP1 junction: one against GI.1 and GI.3, and the other for GII.2 and GII.4. Following optimization of the reaction variables, we standardized the reaction conditions at 65 °C with 6 mM MgSO₄, 1.4 mM dNTPs, 7.5 U WarmStart RTx Reverse Transcriptase, and Bst DNA polymerase at 8 U and 10 U for GI and GII, respectively. Amplification was monitored in real-time using a thermocycler platform to ensure precise quantification and detection. Finally, the assay was evaluated through portable isothermal detection device to test its feasibility in on-site settings. Results: Both assays detected the template down to 10²–10³ copies per reaction and showed high target selectivity, yielding no non-specific amplification across 39 enteric pathogens. These assays enabled prompt detection of GI within 10–12 min and of GII within 12–17 min after the reaction was initiated. Onsite validation reveals all template detection below 15 min, demonstrating its potential feasibility in point-of-care applications. Including the sample preparation time, test results were obtained in less than 1 h. Conclusions: This method is a rapid, reliable, and scalable solution for detecting human norovirus in POCT settings for both clinical diagnosis and public health surveillance. Full article

The Epstein–Barr virus (EBV) is a prevalent virus linked to various diseases, including infectious mononucleosis (IM), nasopharyngeal carcinoma, and Hodgkin’s lymphoma. Over the past few decades, EBV diagnostic strategies have evolved significantly—progressing from traditional serological assays and histopathology to more sensitive and specific molecular techniques such as nucleic acid amplification and high-throughput sequencing (HTS). While conventional methods remain valuable for their accessibility and established clinical use, they are often limited by sensitivity, speed, and multiplexing capability. In contrast, emerging technologies, including isothermal amplification, Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-based diagnostics, multi-omics integration, and AI-assisted analysis, have demonstrated great promise in improving diagnostic accuracy, speed, and applicability in diverse clinical settings, including point-of-care testing (POCT). This review systematically explores the historical development of EBV diagnostic technologies, highlighting key milestones and future trends in precision medicine and global health readiness. Full article

Severe fever with thrombocytopenia syndrome (SFTS) is an emerging tick-borne disease caused by the SFTS virus (SFTSV). A rapid and cost-effective point-of-care testing detection system is important for the early diagnosis of SFTS. Herein, we developed a ready-to-use dried reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for the direct detection of SFTSV in clinical samples. The assay enables simple, RNA-extraction-free detection using heat-treated serum or plasma, followed by a 30 min incubation at 65 °C. The results are visually interpreted through the color emitted, which can be observed under LED light. The established assay demonstrated detection sensitivity for SFTSV at 10⁴ copies/µL and was effective in identifying infections in cats. Despite being less sensitive than real-time RT-PCR, this dried RT-LAMP method offers a rapid, cost-effective alternative suitable for point-of-care use, particularly in remote or resource-limited settings. The simplified workflow and visual readout make it a practical tool for the early detection and daily surveillance of SFTSV in animals. Full article

Mycotoxins are responsible for a multitude of diseases in both humans and animals, resulting in significant medical and economic burdens worldwide. Conventional detection methods, such as enzyme-linked immunosorbent assay (ELISA), high-performance liquid chromatography (HPLC), and liquid chromatography-tandem mass spectrometry (LC-MS/MS), are highly effective, but they are generally confined to laboratory settings. Consequently, there is a growing demand for point-of-care testing (POCT) solutions that are rapid, sensitive, portable, and cost-effective. Lateral flow assays (LFAs) are a pivotal technology in POCT due to their simplicity, rapidity, and ease of use. This review synthesizes data from 78 peer-reviewed studies published between 2015 and 2024, evaluating advances in nanoparticle-based LFAs for detection of singular or multiplex mycotoxin types. Gold nanoparticles (AuNPs) remain the most widely used, due to their favorable optical and surface chemistry; however, significant progress has also been made with silver nanoparticles (AgNPs), magnetic nanoparticles, quantum dots (QDs), nanozymes, and hybrid nanostructures. The integration of multifunctional nanomaterials has enhanced assay sensitivity, specificity, and operational usability, with innovations including smartphone-based readers, signal amplification strategies, and supplementary technologies such as surface-enhanced Raman spectroscopy (SERS). While most singular LFAs achieved moderate sensitivity (0.001–1 ng/mL), only 6% reached ultra-sensitive detection (<0.001 ng/mL), and no significant improvement was evident over time (ρ = −0.162, p = 0.261). In contrast, multiplex assays demonstrated clear performance gains post-2022 (ρ = −0.357, p = 0.0008), largely driven by system-level optimization and advanced nanomaterials. Importantly, the type of sample matrix (e.g., cereals, dairy, feed) did not significantly influence the analytical sensitivity of singular or multiplex lateral LFAs (Kruskal–Wallis p > 0.05), confirming the matrix-independence of these optimized platforms. While analytical challenges remain for complex targets like fumonisins and deoxynivalenol (DON), ongoing innovations in signal amplification, biorecognition chemistry, and assay standardization are driving LFAs toward becoming reliable, ultra-sensitive, and field-deployable platforms for high-throughput mycotoxin screening in global food safety surveillance. Full article

Point-of-care testing (POCT) offers a transformative approach to diagnostics by enabling rapid and accurate results at or near the site of patient care. This is especially valuable in critical care, emergency settings, and resource-limited areas. However, one major limitation of POCT remains its analytical sensitivity, particularly in detecting low concentrations of analytes. To address this, various innovations are being explored, including advanced sensors, signal amplification, and sensitive labels. Among these, bioluminescent proteins have gained attention for their high sensitivity, fast readout, minimal background interference, and simplified instrumentation. Bioluminescence—light emission from biochemical reactions—presents an ideal platform for enhancing POCT sensitivity. In parallel, metal–organic frameworks (MOFs), especially structures like ZIF-8, are emerging as valuable materials in biosensing. Their high porosity, tunable surface properties, and ability to host biomolecules make them excellent candidates for improving analyte capture and signal transduction. When integrated with bioluminescent systems, MOFs can stabilize proteins, concentrate targets, and enhance overall assay performance. This review highlights the role of bioluminescent proteins in medical diagnostics and their application in POCT platforms. We also discuss the potential synergy between MOFs and bioluminescence to overcome current sensitivity limitations. Finally, we examine existing challenges and strategies to optimize these technologies for robust, field-deployable diagnostic tools. By leveraging both the natural sensitivity of bioluminescence and the structural advantages of MOFs, next-generation POCT systems can achieve superior performance, driving forward diagnostic accessibility and patient care outcomes. Full article

In this work, we investigate the fundamental usability of fluorescent DNA-templated silver nanoclusters (DNA-AgNCs) as sensors for Point-of Care-Testing (PoCT) applications. We developed a microfluidic platform for the generation of droplets containing DNA-AgNCs in defined, different chemical environments. The droplets are read out fluorescently at two different emission wavelengths. For the pre-evaluation for the usage of biologically relevant matrices with DNA-AgNCs, the response of two different DNA-AgNCs to a variation in pH and sodium chloride concentration was acquired. Our compact and simple setup can detect DNA-AgNCs well below 100 nM and allows the characterization of the fluorescence response of DNA-based biohybrid nanosensors to changes in the chemical environment within short measurement times. The model DNA-AgNCs remain fluorescent throughout the physiologically relevant chloride concentrations and up to 150 mM. Upon shifts in pH, the DNA-AgNCs showed a complex fluorescence intensity response. The model DNA-AgNCs differ strongly in their response characteristics to the applied changes in their environments. With our work, we show the feasibility of the use of DNA-AgNCs as sensors in a simple microfluidic setup that can be used as a building block for PoCT applications while highlighting challenges in their adaption for use with biologically relevant matrices. Full article

Antithrombin (AT) plays a crucial role in the human anticoagulant system and has extensive clinical applications. However, traditional detection methods often require large sample volumes, complex procedures, and lengthy processing times. Methods: We integrated digital microfluidics technology with AT detection to develop a point-of-care testing (POCT) device that is user-friendly and fully automated for real-time AT testing. Results: This device allows for automation and enhanced adaptability to various settings, requiring only a minimal sample volume (whole capillary blood), thereby omitting steps such as plasma separation to save time and improve clinical testing efficiency. Comparisons with conventional AT activity detection methods demonstrate a high degree of consistency in the results obtained with this device. Conclusion: The AT detection system we developed exhibits significant effectiveness and holds substantial research potential, positioning it to evolve into a clinically impactful POCT solution for AT assessment. Full article

The rising global burden of chronic diseases and cancer in the workplace has intensified the need for accessible, rapid diagnostic strategies within workplace settings. Point-of-care testing (POCT) offers a decentralized solution, providing timely diagnostic insights without the need for centralized laboratory facilities. In the workplace, POCT offers significant advantages for early detection and management of cancer and chronic diseases, improving employee health outcomes and reducing absenteeism. Concurrently, the development of digital health passports has created secure, dynamic platforms for managing and sharing personal health data. This review explores the technological innovations underpinning POCT, examines its application in workplace health screening, and analyzes how integration with the Internet of Things (IoT) and digital health passports can enhance early detection and chronic disease management. The discussion extends to the ethical, regulatory and practical challenges associated with implementation. Furthermore, emerging trends such as artificial intelligence-driven diagnostics, blockchain-enabled data security and wearable biosensors are considered as potential future directions. Together, POCT and digital health passports represent a significant evolution towards proactive, personalized workplace healthcare systems. Full article

Point-of-care testing (POCT) has garnered widespread attention due to its rapid, convenient, and efficient detection capabilities, particularly playing an increasingly pivotal role in medical diagnostics and significantly improving the efficiency and quality of healthcare services. Nanozymes, as novel enzyme-mimicking materials, have emerged as a research hotspot owing to their superior catalytic performance, low cost, and robust stability. This review provides a systematic overview of the fundamental characteristics and classifications of nanozymes, along with various sensing strategies employed in POCT applications, colorimetric, electrochemical, fluorescent, chemiluminescent, and surface-enhanced Raman scattering (SERS)-based approaches. Furthermore, this review highlights innovative designs that enhance the sensitivity and accuracy of POCT across multiple domains, such as biomarker detection, environmental monitoring, and food safety analysis, thereby offering novel perspectives for the practical implementation of nanozymes in point-of-care diagnostics. Finally, this review analyzes current challenges in nanozyme-based POCT systems, including limitations in optimizing catalytic activity, ensuring nanozyme homogeneity, and achieving large-scale production, while proposing future development trajectories. Full article

Background: We propose a framework for determining the stage of periodontitis in a personalized medicine context, building on our previously developed model for periodontitis detection. In this study, we improved the earlier model by incorporating additional components to form a comprehensive system for identifying both the presence and stage of periodontitis. Central to the home-use application is an active-matrix metalloproteinase-8 (aMMP-8) mouth rinse test (cut-off: 20 ng/mL), integrated with software delivered via a mobile application. Methods: First, using all the data, we modeled a single polynomial function to distinguish healthy and stage I periodontitis patients from stage II and III patients. Second, we used an already published periodontitis detection function to separate stage I patients from healthy patients. Third, one more function was created that divided stage II and III patients from each other. All functions were modeled by multiple logistic regression analysis from the patient data, which consisted of 149 adult patients visiting dental offices in Thessaloniki, Greece. Results: The complete model demonstrated a sensitivity of 95.8% (95% CI: 92.1–99.4%) and a specificity of 71.0% (95% CI: 55.0–86.9%) for detecting periodontitis. Among those identified with periodontitis, the correct stage was determined in 61.1% of cases, with stage-specific accuracies of 64.3% for stage I, 60.5% for stage II, and 60.9% for stage III. All testing was performed on patient data with which the complete model was formed. Conclusions: The results of this study showed that with sufficient data and using multiple logistic regression analysis, a model can be created to simultaneously identify the presence and stage of periodontitis. Overall, in the complete model generated, a mouth rinse aMMP-8 test result with a cut-off value of 20 ng/mL, Visible Plaque Index (VPI) and information of patient’s teeth number present were found to be important factors to determine the stage of periodontitis in a personalized medicine manner for everyone to use. Full article