Search Results (562)

Background: Efficient nucleic acid extraction is essential for reliable viral load testing, yet performance can differ widely depending on the extraction system and sample type. We compared three automated platforms, QIAcube, EZ1 Advanced, and Maxwell RSC, for their ability to recover cytomegalovirus (CMV) DNA and West Nile virus (WNV) RNA from common clinical matrices. Methods: Mock specimens were prepared using whole blood, plasma, serum, and urine collected from two donors. Samples were spiked with CMV or WNV culture material and extracted in triplicate on each platform according to the manufacturers’ protocols. Viral loads were measured using ELITech ELITE MGB assays on the InGenius system. Whole blood samples were also tested after a 1:4 dilution. Matrix-specific standard curves were applied, and viral loads were compared using Wilcoxon rank-sum tests with false-discovery rate adjustment. Results: Extraction efficiency differed substantially by platform and specimen type. For CMV, QIAcube consistently produced the highest DNA recovery across all matrices, with particularly large differences in plasma and serum, where EZ1 and Maxwell RSC yielded significantly lower loads. The WNV results varied by matrix: EZ1 and QIAcube performed similarly in plasma, while Maxwell RSC achieved the highest RNA recovery in whole blood. While the QIAcube exhibited reduced WNV recovery in blood, it achieved the best performance in serum, as specified by the kit. No significant platform differences were observed for urine. Diluting whole blood reduced variability between platforms. Conclusions: Both sample matrix and extraction system strongly influence nucleic acid recovery. These results highlight the need for matrix-specific validation and standardized extraction approaches in molecular diagnostics. Full article

Heterogeneous expression of a single surface protein within one cell population can drive major functional differences, yet low-expression subtypes remain difficult to isolate. Conventional tube-based immunomagnetic separation collapses all labelled cells into one positive fraction and thus cannot resolve small differences in marker abundance. Here, we present MagSculptor, a microfluidic platform for high-resolution magnetic fractionation of low-expression EpCAM-defined subtypes within one immunomagnetically labelled population at a time. Arrays of soft-magnetic strips create localized high-gradient zones that map modest differences in bead loading onto distinct capture positions, yielding High (H), Medium (M), Low (L), and Negative (N) fractions. Finite element method simulations of coupled magnetic and hydrodynamic fields quantify the field gradients and define an operating window. Experimentally, epithelial cancer cell lines processed sequentially under identical settings show reproducible subtype partitioning. In a low-EpCAM model (MDA-MB-231), conventional flow cytometry, under standard EpCAM staining conditions, did not yield a robust EpCAM-positive gate, whereas MagSculptor still revealed graded subpopulations. Western blotting confirms a monotonic decrease in EpCAM abundance from H to N, and doxorubicin assays show distinct in vitro drug sensitivities, while viability remains above 95%. MagSculptor thus helps extend immunomagnetic separation from binary enrichment to multi-level isolation of low-expression subtypes and provides a convenient front-end for downstream functional and molecular analyses. Full article

Photocatalytic degradation of organic molecules using TiO₂ has attracted attention in wastewater treatment because it can decompose organic compounds that are difficult to decompose by other methods. Meanwhile, efficient photocatalytic water treatment is difficult because it is not easy to separate nano-sized photocatalysts from water. In this review, we have described two approaches to solve the water separation challenge in the suspension type TiO₂ photocatalysts, which are uniformly distributed in water: magnetic TiO₂/SiO₂/Fe₃O₄ nanoparticles and TiO₂-polystyrene beads. The preparation, characterization, and photocatalytic performance of the two types of photocatalysts and their application are discussed. Finally, we compare two types of photocatalysts while focusing on the respective advantages and disadvantages of each, and the future direction of research. Full article

Photocatalytic water sterilization has emerged as a promising sustainable technology for addressing microbial contamination across diverse sectors including healthcare, food production, and environmental management. This review examines the fundamental mechanisms and recent advances in photocatalytic water sterilization, with a particular emphasis on the differential bactericidal pathways against Gram-negative and Gram-positive bacteria. Gram-negative bacteria undergo a two-step inactivation process involving initial outer membrane lipopolysaccharide (LPS) degradation followed by inner membrane disruption, whereas Gram-positive bacteria exhibit simpler kinetics due to direct oxidative attacks on their thick peptidoglycan layer. Escherichia coli has long been used as the gold standard in photocatalytic sterilization studies owing to its aerobic nature and suitability for the colony-counting method. In contrast, Lactobacillus casei, a facultative anaerobe, can be cultured statically and evaluated rapidly using turbidity-based optical density measurements. Therefore, both organisms serve complementary roles depending on the experimental objectives—E. coli for precise quantification and L. casei for rapid, practical assessments of Gram-positive bacterial inactivation under laboratory conditions. We also describe sterilization using light alone while comparing it to photocatalytic sterilization and then discuss two innovative suspension-based photocatalyst systems: polystyrene bead-supported TiO₂/SiO₂ composites offering balanced reactivity and separability and magnetic TiO₂-SiO₂/Fe₃O₄ nanoparticles enabling rapid magnetic recovery. Future research directions should prioritize enhancing visible-light efficiency using metal-doped TiO₂ such as Cu-doped systems; improving catalyst durability; developing new applications of photocatalysts, such as protecting RO membranes; and validating scalability across diverse industrial and medical water treatment applications. Full article

The detailed characterization of antigen-specific serum antibodies is hindered by the lack of efficient, gentle isolation methods. In this context, standard column affinity chromatography, although a powerful purification tool, presents practical challenges, including high antigen consumption and elution conditions that risk inducing antibody polyreactivity, while conventional acidic elution often compromises antibody integrity. This study introduces a novel microscale method for isolating specific immunoglobulins using anionic detergents as mild eluents. We employed antigen-functionalized hydrogel microarrays and magnetic beads as micro-immunosorbents. Among the tested detergents, sodium lauroyl glutamate (SLG) was optimal, achieving up to 78.3% recovery of functional antibodies. The optimized protocol, including recovery via G25-Sephadex gel filtration, effectively isolated specific antibodies from complex serum, retaining 58.5–85.3% of their functional bioactivity. Multiplex immunoassays confirmed the high specificity of the isolated antibodies and the lack of detergent-induced polyreactivity. The method was successfully adapted to isolate both specific antibodies (virus, dietary, and autoimmune) and total IgG, demonstrating versatility across platforms. This work establishes a robust, efficient, and gentle workflow for obtaining high-purity, bioactive antibodies, enabling their subsequent in-depth analysis for research applications. Full article

Acute myocardial infarction (AMI) is a rapidly progressing cardiovascular condition associated with high mortality. Myoglobin is an early biomarker of AMI; however, its detection using conventional methods is limited by complex workflows and low resistance to interference. In this study, we developed an integrated myoglobin detection platform that combined magneto-immunoassay with microfluidic technology. A giant magnetoresistance (GMR) sensor was fabricated using micro-electro-mechanical systems (MEMS). The designed microfluidic chip integrated sample pretreatment, immunoreaction, and magnetic signal capture functionalities. Its built-in GMR sensor, labeled with magnetic nanoparticles, directly converted the “antigen–antibody” biochemical signal into detectable magnetoresistance changes, thereby enabling the indirect detection of myoglobin. A magneto-immunoassay analysis system consisted of a fluidic drive, magnetic field control, and data acquisition functions. Various key parameters were optimized, including EDC/NHS concentration, antibody concentration, and magnetic bead size. Under the optimal conditions and using 1 μm magnetic beads as labels and an external detection magnetic field of 60 Oe, the platform successfully detected myoglobin at 75 ng/mL with ∆MR ≥ 0.202%. Specificity tests demonstrated that the platform had high specificity for myoglobin and could effectively distinguish myoglobin from bovine serum albumin (BSA) and troponin I. This study presents a rapid, accurate myoglobin detection platform that can be applied for the early diagnosis of AMI. Full article

Mimotope-based immunoassays offer an eco-friendly alternative to chemically synthesized antigens for the quantitative analysis of small molecules, but their use for practical on-site and high-throughput residue monitoring remains limited. Herein, we report the selection, production, and application of a phage display–derived mimotope targeting an anti-uniconazole monoclonal antibody (UCZ-mAb), with the aim of developing two complementary immunoassays that enable sensitive, eco-friendly detection of UCZ residues in agricultural samples. A 12-mer phage-displayed peptide library was screened to identify UCZ-specific mimotopes, and a selected sequence was genetically fused to SpyTag and expressed in Escherichia coli to generate a SpyTagged mimotope. Leveraging the SpyCatcher/SpyTag self-assembly system, the SpyTagged mimotope was directionally conjugated onto SpyCatcher-functionalized time-resolved fluorescence beads (TRFBs) and subsequently used as a signal-labeled competitive antigen in a lateral flow immunoassay (LFIA) designed for rapid on-site screening. In parallel, a wash-free magnetic separation immunoassay (MSIA) suitable for green, high-throughput screening in routine laboratories was established using self-assembled mimotope-TRFB probes. The LFIA and MSIA exhibited half-maximal inhibitory concentrations (IC₅₀) of 3.70–6.72 μg/kg and 16.4–18.3 μg/kg, respectively, in real samples. Spiked-sample recoveries ranged from 91.1 to 107.8% for LFIA and 92.6–115.7% for MSIA, demonstrating acceptable accuracy and precision. These results indicate that the SpyTagged mimotope–based LFIA and MSIA provide complementary, reliable, and sensitive platforms for on-site screening and high-throughput monitoring of UCZ residues in agricultural samples, while avoiding the drawbacks associated with traditional chemical antigen synthesis. Full article

Background: Detecting low-frequency mutations is crucial for predicting prognosis and monitoring minimal residual disease (MRD) in acute myeloid leukemia (AML). However, the presence of abundant wild-type sequences hinders the detection of rare mutant alleles. We present a highly sensitive method called ACE (Amplifying–Cleaving–Enriching) to selectively enrich mutant sequences. Methods: ACE includes three steps: (1) initial PCR amplification using biotin-labeled primers, (2) cleavage of wild-type sequences with a specific restriction enzyme, and (3) enrichment of undigested mutant alleles via streptavidin-labeled magnetic beads. Results: Using two rounds of ACE, we achieved over 80,000-fold enrichment of mutant sequences carrying FLT3-TKD, enabling the detection of mutant alleles at levels as low as 0.0001% in AML patient blood samples. Additionally, the ACE method can be adapted to nearly any driver mutation by introducing wild-type-specific restriction sites through PCR with mismatched primers, which has been validated in the IDH1 mutation. Furthermore, the ACE method can be flexibly integrated into conventional detection techniques including Sanger sequencing, quantitative real-time PCR, allele-specific PCR, and even with advanced techniques like droplet digital PCR. Conclusions: ACE significantly enhances the sensitivity of existing techniques for rare mutation detection and holds potential for broad clinical applications. Full article

Background: The roles of cytokines and chemokines in the pathogenesis of Chagas cardiomyopathy (CC) have been proposed, yet their clinical significance with respect to conduction disturbances and left ventricular ejection fraction (LVEF) remains unclear. Aim: The objective of this study was to analyze the associations between cytokine levels and systolic function, comparing patients with preserved and reduced ejection fractions. As a secondary objective, we evaluated whether differences were present in cytokine levels within the subgroup with preserved ejection fraction, depending on the presence or absence of intraventricular conduction disturbances. Methods: We conducted an analytical cross-sectional study involving patients with Chagas disease and a healthy control group. Among patients with Chagas disease, those with preserved (>50%) and reduced (<35%) left ventricular ejection fraction (LVEF) were selected. The preserved-LVEF group included individuals with and without conduction disorders. Cytokines (IFN-γ, IL-1β, IL-6, IL-10, IL-12p70, IL-15, IL-17A, MCP-1, MIP1α, TNF-α, and IL-2) were quantified using a magnetic bead-based multiplex assay. Results: Forty-four patients with CD (26 men, 59%) and 14 seronegative controls were included. In the CD group, 50% (n = 22) had preserved LVEF (LVEF > 50%), and 50% (n = 22) had decreased LVEF (≤35%). No significant differences in cytokine concentrations were observed between patients with preserved and reduced LVEF for TNF-α (19.74 ± 8.32 vs. 22.23 ± 6.40 pg/mL; p = 0.189), IL-6 (2.17 ± 2.41 vs. 5.40 ± 6.40 pg/mL; p = 0.145), IL-2 (2.61 ± 1.05 vs. 2.97 ± 1.79 pg/mL; p = 0.481), MCP-1 (214.18 ± 96.99 vs. 183.83 ± 63.21 pg/mL; p = 0.481) and IFN-γ (9.04 ± 4.90 vs. 7.64 ± 3.78 pg/mL; p = 0.372). Within the subgroup with preserved LVEF (n = 22), those with conduction disorders (n = 10) exhibited higher levels of IL-10 (24.49 vs. 9.83 pg/mL; q = 0.009), IL-12p70 (13.20 vs. 9.02 pg/mL; q = 0.027), IL-2 (2.70 vs. 2.07 pg/mL; q = 0.023), IL-15 (7.09 vs. 3.36 pg/mL; q = 0.018), MIP1α (10.33 vs. 3.35 pg/mL; q = 0.014) and IFN-γ (10.83 vs. 7.25 pg/mL; q = 0.005), compared to those without conduction disorders (n = 12). Notably, patients with CD and preserved LVEF (>50%) without conduction disturbances presented cytokine profiles similar to those of seronegative healthy controls with LVEF ≥ 50%. Conclusions: Elevated levels of specific cytokines were associated with conduction disturbances in patients with preserved LVEF. Despite this finding, a causal relationship cannot be established, and future studies are needed to explore their prognostic or therapeutic significance. Full article

Additive manufacturing (AM) offers significant potential but faces challenges in controlling rapid solidification processes due to thermal conditions. The application of magnetic fields provides a promising path to influence liquid metal behavior during solidification. Thermoelectromagnetic convection (TEMC) is one of the mechanisms by which an applied static magnetic field can induce melt flow, where thermal gradients at the solid–liquid interface generate thermoelectric currents, and in the presence of an external magnetic field induce Lorentz force that drives liquid convection, leading to enhanced heat transfer. This study investigates the impact of moderate static magnetic fields on the laser melting process of a Sn-10%wt.Pb alloy. It is found that applying a magnetic field significantly widens and deepens laser weld beads. Bead depth and width under different field strengths and orientations are measured. Numerical models are developed to calculate the TEMC current distribution and flow in the melt pool. Full article

Backgroud/Objectives: Canine Visceral Leishmaniasis (CVL), caused by Leishmania infantum, is a significant public health concern due to dogs serving as reservoirs for human infection. An accurate and rapid diagnostic method to distinguish symptomatic and asymptomatic CVL from healthy and vaccinated animals is essential for controlling canine and human disease. Developing innovative antibody detection techniques and exploring new antigens are essential for enhancing CVL testing efficiency. Our study focuses on a multiplex flow cytometry technique to detect Leishmania-specific antibodies in canine serum. This involved conjugating small peptides with carrier proteins or peptide tags, sequences designed to facilitate bead coupling. Methods: A peptide from the L. infantum A2 protein was coupled to beads in three forms: unconjugated, conjugated with BSA, and conjugated with a C-terminal β-alanine–lysine (x4)–cysteine TAG. This TAG was previously designed to enhance peptide solubility, improve binding efficiency, and provide functional groups for covalent attachment to the beads, ensuring stable immobilization in the multiplex assay. Results: Our results suggest that the multiplex approach shows promise as a rapid serological test for CVL, particularly with TAG-conjugated peptides, which optimize bead coupling. However, peptide/BSA conjugation revealed anti-BSA antibodies in samples from healthy and CVL dogs. Conclusions: In conclusion, our findings highlight the potential of multiplex methodologies to enhance CVL diagnostics and caution against using BSA as a bead coupling agent in serological tests for canine samples due to its impact on test specificity and sensitivity. Full article

Background/Objectives: Ergot alkaloids are mycotoxins produced mainly by fungi of the genus Claviceps, infecting a wide variety of plants, especially cereals. These toxins usually manifest as black, hardened sclerotia (ergots), though they may also be invisible when dispersed in grain. They pose a significant risk to animals and humans when present in contaminated cereals. They can cause ergotism, with vasoconstriction, ischemia, hallucinations, and in severe cases gangrene. This study was carried out in response to the European legislative actions which determine the permissible levels of ergot alkaloids in cereals. Historically, consumers manually removed visible sclerotia from grain, and farmers applied fertilizers or timed harvests to specific periods to mitigate contamination. However, these traditional methods have proven insufficient. We therefore explored advanced techniques for detecting and quantifying ergot-contaminated cereals, as well as methods for reducing ergot alkaloid concentrations. Methods: Searches were conducted in scientific databases including Google Scholar, PubMed, and Scopus to identify research articles, reviews, and experimental studies published mainly between 2012 and August 2025, including accepted or in-press manuscripts, with special attention to works from 2021 onward to capture the most recent advancements. Results/Conclusions: Ultra-high-performance liquid chromatography–tandem mass spectrometry (UHPLC-MS/MS) is the reference method for confirmatory, epimer-aware quantification of ergot alkaloids, and is already standardized. Recent QuEChERS-UHPLC-MS/MS workflows in cereal matrices, including oat-based products, routinely achieve limits of quantification of about 0.5–1.0 µg/kg with single-run analysis times of about 5–15 min. Rapid screening options complement, rather than replace, confirmatory mass spectrometry: magnetic bead-based immunoassays that use magnetic separation and a smartphone-linked potentiostat provide sub-hour turnaround and field portability for trained quality-assurance staff, although external validation and calibration traceable to LC-MS/MS remain prerequisites for routine use. In practice, operators are adopting tiered, orthogonal workflows (e.g., immunoassay or electronic-nose triage at intake followed by DNA-based checks on grain washings and LC–MS/MS confirmation, or hydrazinolysis “sum parameter” screening followed by targeted MS speciation). Such combinations reduce turnaround time while preserving analytical rigor. Biotechnology also offers potential solutions for reducing ergot alkaloid concentrations at the source. Finally, to enhance consumer safety, artificial intelligence and blockchain-based food traceability appear highly effective. These systems can connect all stakeholders from producers to consumers, allowing for real-time updates on food safety and rapid responses to contamination issues. This review primarily synthesizes advances in analytical detection of ergot alkaloids, while mitigation strategies and supply chain traceability are covered concisely as supporting context for decision making. Full article

We previously developed a bead-coupled ligase detection reaction (LDR) assay that enables simple and rapid detection of single-nucleotide variations (SNVs) using synthetic oligonucleotide templates. In the present study, this approach was extended to genomic DNA extracted from colorectal cancer cell lines to evaluate its applicability to clinically relevant samples. Targeting codon 12 of the KRAS gene, PCR-amplified products served as templates for bead-coupled LDR, and fluorescence excitation–emission matrix (EEM) analysis was employed for signal readout. The four fluorophores used in the assay exhibited distinct spectral properties, allowing their signals to be clearly resolved within the EEM profiles. This mapping provided characteristic fluorescence signatures that revealed the underlying genotypes, enabling not only the distinction between homozygous and heterozygous states but also the precise identification of allele compositions, as exemplified by G/A, T/T, G/G, and G/C in colorectal cancer cell lines. The single-tube workflow, integrating magnetic bead capture with fluorescence-based detection, demonstrated robustness, speed, and cost-effectiveness compared with conventional mutation detection methods. These findings confirm that the LDR–EEM platform can be successfully applied to genomic DNA analysis, underscoring its potential as an accessible and reliable tool for SNV detection in both research and diagnostic contexts. Full article

The epidemic of infectious diseases, such as influenza A, has imposed a severe health burden on the population. Early detection, diagnosis, reporting, isolation, and treatment are crucial for the prevention, control, and management of infectious diseases. Nucleic acid testing represents a vital approach for the rapid diagnosis of pathogenic microorganism types. However, current nucleic acid detection methods face notable bottlenecks: traditional CRISPR fluorescence assays require time-consuming pre-amplification of target nucleic acids, while existing carbon-nanotube field-effect transistor (FET)-based platforms, though amplification-free, often necessitate complex chip surface modification and probe immobilization, and suffer from non-reusable chips, all limiting their utility in point-of-care testing (POCT) and large-scale screening. This study reports a CRISPR-based amplification-free RNA detection platform (CRISPR-FET) for the rapid identification of influenza A virus. The CRISPR-FET platform described herein enables the detection of viral RNA without amplification within 20 min, with a limit of detection as low as 1 copy/μL. Secondly, a reporter RNA conjugated with gold particles is used to achieve signal amplification in FET detection; meanwhile, the method eliminates probe immobilization, thereby omitting this step and simplifying chip modification to reduce complex work-flows and pre-treatment costs. The chip’s reusability further enhances cost-effectiveness. Additionally, streptavidin-modified magnetic bead adsorption minimizes background errors from excessive reporter RNA and non-target nucleic acids. Finally, validation with 24 clinical samples confirmed the platform’s efficacy. By integrating rapidity, simplicity, and high sensitivity, alongside cost advantages from reusable chips, this CRISPR-FET platform meets the critical need for early influenza A diagnosis and holds promise for advancing POCT and large-scale epidemiological screening. Full article

Alcohol’s chronic neurotoxic and degenerative effects mediate alcohol-related brain damage (ARBD), which is marked by neurobehavioral, cognitive, and motor deficits. Major underlying abnormalities include impairments in signaling through the insulin and insulin-like growth factor (IGF) pathways, which regulate energy metabolism. This study examined the potential role of dysregulated incretin network-related mechanisms as mediators of ARBD and evaluated a non-invasive serum exosome (S-EV)-based approach for detecting brain abnormalities. Frontal lobe tissue and S-EVs isolated from Long–Evans adolescent rats maintained for 2 weeks on control or 24% ethanol (caloric) containing liquid diets (n = 8/group) were analyzed using multiplex magnetic bead-based enzyme-linked immunosorbent assays (ELISAs). ARBD was associated with significantly reduced insulin, C-peptide, glucagon, ghrelin, leptin, GIP, and amylin levels in the frontal lobe and/or S-EV samples. In contrast, chronic ethanol exposure had no significant effects on PP, PYY, or GLP-1, and it did not increase proinflammatory cytokine expression. Chronic ethanol feeding broadly affected (primarily inhibiting) the expression of metabolic hormones linked to insulin/IGF signaling. The reductions in GIP and amylin suggest potential targets for therapeutic intervention to enhance brain energy metabolism via insulin networks. On the other hand, the findings suggest that GLP-1 receptor agonists may have limited efficacy in remediating the effects of ARBD. Finally, the results support the use of non-invasive S-EV assays to detect and guide treatment for metabolic brain dysfunction in ARBD. Full article