Search Results (1,530)

Preclinical models play a critical role in the development of PSMA-targeted radiopharmaceuticals for prostate cancer. However, many existing models have practical limitations, including slow tumor growth, low engraftment rates, and restricted availability, and all human PSMA-positive lines are confined to immunodeficient hosts. We developed and characterized a novel PSMA-expressing transgenic cell line, CT26-PSMA, as a practical tool for preclinical screening of PSMA-targeting agents. The CT26-PSMA cell line was established by stable transfection of the murine colon carcinoma CT26 cell line with human PSMA using the Sleeping Beauty transposon system. PSMA expression was confirmed by RT-qPCR (reverse transcription quantitative polymerase chain reaction), flow cytometry, and radioligand saturation binding on intact cells. Two [⁶⁸Ga]Ga-labeled radioconjugates—the well-established PSMA-617 and a newly synthesized conjugate (Conjugate-1)—were used to validate the functionality of the model through in vitro binding, uptake and internalization studies, and through ex vivo biodistribution in CT26-PSMA tumor-bearing athymic male nu/nu mice. The CT26-PSMA cell line demonstrated high and stable PSMA expression, with approximately 95% of cells expressing the biomarker and no measurable loss over 16 passages in antibiotic-free medium. Saturation binding gave a receptor density of ∼3.5 × 10⁶ sites per cell, approximately four-fold higher than that of LNCaP cells (∼0.8 × 10⁶), with dissociation constants that were indistinguishable between the two radioconjugates and between the two cell lines (K_d 9.0–11.6 nM). Subcutaneous tumors reached ~300 mm³ within 8–10 days of inoculation, with a take rate of 10/10 versus 1/10 for LNCaP (Fisher’s exact test, p = 1.2 × 10⁻⁴). Both radiotracers showed saturable, 2-PMPA-blockable binding and uptake in CT26-PSMA cells, confirming the functional activity of the recombinant receptor. Biodistribution studies revealed accumulation of both conjugates in CT26-PSMA tumors, with generally comparable tumor-to-background profiles. The CT26-PSMA cell line represents a robust, rapid, and reproducible platform for preclinical evaluation of PSMA-targeting radiopharmaceuticals, and its murine BALB/c origin permits engraftment in immunocompetent or minimally immunosuppressed hosts, whereas existing human PSMA-positive lines do not. It is intended as a screening platform rather than as a model of prostate cancer biology. The validation data obtained with [⁶⁸Ga]Ga-labelled conjugates confirm the suitability of this cell line for future studies of PSMA-directed compounds. Full article

Soybean root rot caused by Fusarium graminearum is an important soil-borne disease. It hinders seedling establishment and ultimately reduces soybean yield. Resistant germplasm and reliable molecular markers are therefore needed for resistance breeding. In this study, 336 soybean accessions were evaluated for resistance to F. graminearum root rot using the disease severity index (DSI), which ranged from 5.71 to 100.00 across the association panel. Genome-wide association analysis was performed using resequencing-based single nucleotide polymorphism (SNP) data with mixed linear model (MLM) and Fixed and random model Circulating Probability Unification (FarmCPU) models, which detected 117 and 113 candidate resistance-associated SNPs, respectively. Among these, 105 shared SNPs were used to define candidate genomic intervals containing 247 annotated genes. Based on Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment, functional annotation, and allelic-effect analysis, six candidate genes and their associated exonic SNPs were prioritized. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis showed infection-responsive expression patterns for all six candidate genes, with Glyma.17g202500 and Glyma.18g266700 showing stronger induction in the resistant accession. Two SNPs in these genes were converted into Kompetitive allele-specific PCR (KASP) assays. KASP-S17_32244510 and KASP-S18_55105706 were successfully developed for genotype screening, with screening efficiencies of 73.08% and 74.29%, respectively. These findings identify useful genetic targets and molecular markers for improving soybean resistance to root rot caused by F. graminearum. Full article

Background: Staphylococcus aureus causes bloodstream and skin infections in humans. The prevalence of multidrug-resistant S. aureus strains means new antibiotics are needed. A novel antimicrobial drug named SK-03-92, a synthetic aromatic organic stilbenoid compound, kills S. aureus cells within 30 min, but an increase in both biofilm formation and persister cells occurs. SK-03-92 treatment downregulates transcription of the biofilm regulating protein regulator (brpR) gene and biofilm regulating protein sensor (brpS) gene in S. aureus. BrpR/BrpS system may be a LytTR regulatory system tied to biofilm formation, creation of persister cells, and late-stage competence in S. aureus. The aim of this study was to determine what biofilm, late-stage competence, and persister-associated genes were regulated in a brpR mutant compared to wild-type strains. Methods: In this study, involvement of BrpR in regulating other genes was assessed by comparing transcriptional changes in a brpR mutant strain to the S. aureus parent strain via RNA sequencing (RNA-Seq). Bioinformatic analysis was then performed on the RNA-Seq data to assess what biochemical pathways might be involved. Results: From these analyses, 440 genes were identified that had significant differences in transcript abundance when comparing the brpR mutant to wild-type strains. Quantitative reverse transcription polymerase chain reaction analysis confirmed bacA, icd, metE, and pdhA transcript levels were lower, whereas alr and mraY were higher in the brpR mutant versus wild-type strain. Furthermore, an enzymatic assay targeting NADH production from the pyruvate dehydrogenase complex showed lower levels in the mutant compared to wild-type strain. Conclusions: Overall, the study demonstrated several biosynthetic pathways tied to biofilm formation and late-stage competency may be regulated by BrpR and some potential leads for the mechanism of action of the SK-03-92 drug were uncovered. Full article

Background/Objectives: At-risk metabolic dysfunction-associated steatohepatitis (MASH) is associated with increased risks of cirrhosis, hepatocellular carcinoma, and liver-related mortality. Because impaired efferocytosis contributes to persistent hepatic inflammation and fibrotic remodeling in MASH, we investigated whether efferocytosis-associated molecular signatures could identify at-risk MASH. Methods: Bulk RNA-sequencing datasets from Gene Expression Omnibus (GSE135251 and GSE174478) were analyzed to identify differentially expressed efferocytosis-related genes and characterize associated pathways and immune infiltration patterns. Machine learning-based feature selection was used to identify hub genes, which were incorporated into a transcriptomic nomogram. Experimental validation involved reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and Western blotting of liver tissues from a Western diet-induced murine metabolic dysfunction-associated steatotic liver disease (MASLD) model. Plasma proteomic data were analyzed to explore the discriminatory performance of hub gene products. Results: A total of 17 efferocytosis-related genes (ERGs) associated with at-risk MASH were identified and enriched in pathways related to efferocytosis, inflammation, and immune regulation. Five hub genes, CD24, CHI3L1, TREM2, PTGS2, and LGR6, were shared by all three feature-selection approaches and significantly upregulated in at-risk MASH. A transcriptomic nomogram yielded area under the curve (AUC) values of 0.866 and 0.805 in the training and external cohorts, respectively. In the murine MASLD model, all five hub genes showed increased mRNA and protein expression in advanced disease. Exploratory plasma proteomic analysis identified elevated circulating TREM2 and CHI3L1 levels in at-risk MASH, and a simplified plasma-based model achieved an AUC of 0.736. Conclusions: This study identified a five-gene efferocytosis-associated signature and developed models for identifying at-risk MASH, suggesting that these genes warrant further evaluation as candidate biomarkers. Full article

Cysticercosis, caused by the larval stage of Taenia solium (Cysticercus cellulosae), is a neglected tropical disease threatening public health. Thioredoxin peroxidase (TPx) is a key antioxidant protein secreted by the parasite, but its role in macrophage polarization remains unclear. In this study, THP-1-derived macrophages were treated with TPx protein for 24 h and 48 h. Flow cytometry, reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and Western blot were employed to assess reactive oxygen species (ROS) levels, M1/M2 cell proportions, mRNA expression of tumor necrosis factor-alpha (TNF-α) and interleukin-10 (IL-10), and protein expression of inducible nitric oxide synthase (iNOS) and arginase-1 (Arg-1). Transcriptome sequencing was performed to screen for signaling pathways, and enzyme-linked immunosorbent assay (ELISA) was subsequently used to measure cGMP levels. The PKG inhibitor KT-5823 was used for functional validation. The results show that TPx significantly increased the proportion of M1 macrophages from 4.58% to 9.24% at 24 h, and promoted M2 macrophages from 4.12% to 6.87% at 48 h, while ROS levels decreased to 0.80-fold at 48 h (p < 0.05). RT-qPCR revealed that TPx markedly upregulated TNF-α (1.62-fold) at 24 h and IL-10 (1.52-fold) at 48 h (p < 0.05). Western blot showed that TPx increased iNOS expression by 2.55-fold at 24 h and Arg-1 expression by 2.01-fold at 48 h. KEGG analysis revealed upregulation of the cGMP-PKG pathway at 48 h, with notably increased cGMP content (1.48-fold) and PKG expression (1.86-fold) (p < 0.05). Furthermore, KT-5823 pretreatment effectively reversed TPx-induced Arg-1 upregulation (from 1.66-fold to 1.02-fold, p < 0.05). These findings demonstrate that Cysticercus cellulosae TPx induces M1 polarization at 24 h and promotes M2 polarization at 48 h through activation of the cGMP-PKG signaling pathway, thereby facilitating immune evasion. Full article

Classical swine fever (CSF) is a highly contagious transboundary animal disease that causes major economic losses to the global swine industry and remains notifiable to the World Organisation for Animal Health (WOAH). Handling and transport of infectious classical swine fever virus (CSFV)-positive samples require biosafety level 3 (BSL-3) containment, limiting diagnostic capacity and interlaboratory exchange in many regions. In this study, we evaluated the efficacy of PrimeStore^® Molecular Transport Medium (PS-MTM) for eliminating detectable CSFV infectivity while preserving viral ribonucleic acid (RNA) detectability for downstream molecular detection. Validation assays were performed using the CSFV Alfort/187 reference strain and clinical samples collected from pigs experimentally infected with the Catalonia01 and Margarita CSFV strains. Residual infectivity was assessed by virus isolation, whereas viral RNA detection and stability were evaluated using the WOAH-recommended real-time reverse transcription quantitative polymerase chain reaction(RT-qPCR) assay. Following PS-MTM treatment, no residual infectivity was detected in either cell culture-derived virus stocks or clinical samples, including extensive replicate testing and three subsequent blind passages. RT-qPCR analyses confirmed preservation and stability of detectable CSFV RNA for at least 60 days under the storage conditions evaluated, including refrigerated storage and room temperature storage for Catalonia01-derived samples. Overall, these findings demonstrate that PS-MTM eliminates detectable CSFV infectivity while preserving viral RNA for molecular diagnosis. The implementation of PS-MTM may facilitate safer handling, transport, and interlaboratory exchange of CSFV-positive samples, contributing to improved biosafety, diagnostic harmonization, and global surveillance capacity for this transboundary animal disease. Full article

Intensive training induces redox and inflammatory signaling that may facilitate adaptation but can also reflect excessive physiological strain. In this exploratory pilot longitudinal study, we assessed whether 8-iso-prostaglandin F2α (8-iso-PGF2α), a lipid peroxidation marker, and transcripts related to arachidonic acid lipoxygenase pathways change during a 10-week preparatory training period in elite female volleyball players. Twelve professional athletes were sampled at baseline after a training break and after training completion. 8-iso-PGF2α was quantified by gas chromatography–mass spectrometry (GC-MS), and the relative expression of arachidonate 5-lipoxygenase (ALOX5), arachidonate 12-lipoxygenase (ALOX12), and arachidonate 15-lipoxygenase (ALOX15) genes was measured by reverse transcription quantitative polymerase chain reaction (RT-qPCR) in leukocytes and plasma-derived RNA. Training increased 8-iso-PGF2α from 0.06 [0.05–0.09] to 0.16 [0.08–0.22] ng/mL (p = 0.03). ALOX5 expression increased in leukocytes (0.80 ± 0.29 vs. 1.00 ± 0.29; p = 0.02) and plasma-derived RNA (0.03 [0.02–0.06] vs. 0.06 [0.05–0.11]; p = 0.04), whereas no statistically detectable pre-to-post changes were observed for ALOX12 and ALOX15 expression. Given the exploratory design and small sample size, these results should be interpreted as hypothesis-generating. The observed changes suggest that 8-iso-PGF2α and ALOX5 are associated with physiological responses to preparatory training and exercise-induced lipid oxidative remodeling; however, their potential utility for monitoring training adaptation requires validation in larger, prospectively designed controlled cohorts. Full article

Background/Objectives: Mesoporous silica nanoparticles (MSNs) are widely investigated as nanocarriers for drug delivery, gene transfer, and bioimaging. However, the mechanisms underlying their size-dependent cytotoxicity at the metabolic level remain incompletely understood. This study aimed to determine whether different-sized MSNs induce distinct patterns of subcellular injury and metabolic reprogramming in lung epithelial cells. Methods: A549 cells were exposed to 80 nm and 600 nm MSNs at 50 and 200 μg/mL for 24 h. Ultrastructural changes were examined by transmission electron microscopy (TEM). Intracellular reactive oxygen species (ROS) and Ca²⁺ were measured by 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA) and Fluo-4 AM fluorescence, respectively. Inflammatory gene expression (IL1B, IL6, TNFA, HIF1A) was quantified by reverse transcription quantitative polymerase chain reaction (RT-qPCR). Untargeted metabolomics were performed using combined gas chromatography–mass spectrometry (GC-MS) and liquid chromatography–mass spectrometry (LC-MS) platforms, followed by principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), and MetaboAnalyst-based pathway enrichment. Results: TEM revealed distinct size-dependent subcellular distributions: 80 nm MSNs were predominantly associated with mitochondrial abnormalities, including cristae disruption, swelling, and mitophagy-like features, whereas 600 nm MSNs accumulated in endocytic vesicles with membrane disruption. Metabolomic profiling showed that 80 nm MSNs were associated with TCA cycle blockade—characterized by the accumulation of early intermediates (citrate, oxaloacetate) and the depletion of distal intermediates (fumarate, malate)—with compensatory glycolytic activation (increased glyceraldehyde-3-phosphate and pyruvate) and reduced deoxynucleotide pools (dCDP, dUMP). By contrast, 600 nm MSNs triggered broad nucleotide triphosphate accumulation (ATP, CTP, dGTP, dCTP), amino acid depletion, and robust inflammatory activation, including a ~136-fold increase in IL1B expression and HIF1A transcriptional upregulation. PCA and PLS-DA confirmed distinct size-dependent metabolic phenotypes. Conclusions: MSN size strongly influences subcellular targeting—80 nm particles were predominantly associated with mitochondrial injury while 600 nm particles disrupted endocytic vesicles—driving qualitatively distinct patterns of metabolic reprogramming and inflammatory signaling. These findings establish a correlative mechanistic framework linking particle size to organelle-specific injury and provide candidate metabolic markers for nanotoxicological evaluation. Full article

Background: Cyclophosphamide (CTX)-induced immunosuppression involves systemic toxicity, splenic atrophy, and broad transcriptomic disruption. Icariin (ICA) has immunomodulatory activity, but the systems-level features associated with splenic recovery remain incompletely defined. Methods and Results: Male mice were assigned to control, CTX, and CTX plus ICA (20, 40, or 80 mg/kg) groups. Phenotypic measurements were integrated with splenic RNA sequencing, differential expression analysis, targeted gene-set analysis, and weighted gene co-expression network analysis (WGCNA). ICA produced dose-associated improvement in spleen index and body weight trajectory, with the most consistent phenotypic response at 80 mg/kg. Overall transcriptomic separation was supported by permutational multivariate analysis of variance (PERMANOVA) (F = 17.18, R² = 0.873, p < 0.001). WGCNA identified a recovery-associated turquoise module; Myb was assigned to this module, whereas Gata1 belonged to a distinct royalblue module. Complement and chemokine gene sets provided the strongest targeted enrichment evidence, and reverse transcription quantitative polymerase chain reaction (RT-qPCR) confirmed dose-associated changes in selected innate immune transcripts. Marker-based lineage signatures indicated non-uniform recovery, including persistent depression of the B-cell signature. Conclusion: ICA-associated phenotypic recovery coincided with partial, non-uniform remodeling of splenic transcriptional programs. The Myb- and Gata1-associated findings are hypothesis-generating co-expression signals and do not establish transcription-factor binding or causality. Full article

Dengue virus (DENV) remains a major global public health challenge, driven by the expanding distribution of Aedes vectors, rapid urbanization, and increasing climate variability. This review aimed to synthesize current evidence on dengue transmission dynamics, immunopathogenesis, molecular epidemiology, diagnostic and therapeutic advances, vaccine development, and the influence of climatic factors on disease patterns. A comprehensive search of major electronic databases was conducted to identify relevant literature, followed by a qualitative synthesis of the evidence. The evidence highlights complex transmission cycles involving Aedes aegypti and Aedes albopictus, with transmission strongly influenced by temperature, humidity, rainfall, and extreme climate events. Advances in immunopathogenesis research have improved understanding of mechanisms associated with severe disease, including antibody-dependent enhancement and dysregulated inflammatory responses. Diagnostic innovations, such as reverse transcription polymerase chain reaction (RT-PCR), NS1 antigen detection, and point-of-care technologies, have enhanced case identification, while prevention strategies increasingly incorporate integrated vector management, digital surveillance systems, and emerging vaccines. Integrating epidemiological, molecular, and climatic information may strengthen early warning systems, improve outbreak prediction, and support more effective dengue prevention and control strategies. Full article

Phenolic acids and flavonoids are major bioactive constituents of Sparganium stoloniferum tubers (SL), yet their spatial organization within this medicinal organ remains insufficiently characterized, limiting understanding of how anatomical structure relates to metabolite distribution. To address this, we integrated mass spectrometry imaging (MSI), non-targeted LC–MS and targeted LC–MS/MS metabolomics, and reverse transcription quantitative polymerase chain reaction (RT–qPCR) analysis to characterize the spatial patterns of phenylpropanoid- and auxin-related metabolites and selected transcripts in SL. MSI and non-targeted LC–MS profiling showed that phenylpropanoid- and flavonoid-related metabolites were preferentially accumulated in the peripheral cortex rather than in the stele, although the stele constitutes the dominant internal tissue of the tuber. Targeted LC–MS/MS further confirmed that representative hydroxycinnamic acids and caffeoylquinic acid derivatives were enriched in the cortex and, for most validated metabolites, in the cell wall-enriched fraction. Notably, spatial metabolomic profiling also revealed a contrasting stele-biased distribution of indole-related metabolites, including indole-3-acetamide-related features detected by MSI and non-targeted LC–MS and indole-3-acetic acid (IAA) enrichment validated by targeted LC–MS/MS. RT–qPCR analysis showed that phenylpropanoid biosynthetic genes were generally more highly expressed in the cortex and cell wall-enriched fraction, whereas auxin-related genes showed higher expression in the stele. Together, these results show cortex-biased phenylpropanoid accumulation, preferential association of most validated phenylpropanoid-related metabolites with the cell wall-enriched fraction, and stele-associated IAA accumulation and auxin-related transcript expression. This study provides a spatial framework for understanding metabolite partitioning in medicinal aquatic storage organs and highlights the importance of integrating anatomical, metabolomic, and gene expression information in medicinal plant research. Full article

VERO cells, derived from the kidney epithelium of the African green monkey, are widely used in virology, but their ability to metabolize xenobiotics is not fully understood. Since cytochrome P450 (CYP) enzymes participate in xenobiotic metabolism, we investigated which CYP genes are expressed in VERO-E6 cells. Reverse transcription–quantitative polymerase chain reaction (RT-qPCR) showed that VERO-E6 cells express CYP3A4, CYP3A5, and CYP3A7. In contrast, CYP1A1, CYP1A2, CYP1B1, CYP2E1, CYP2D6, and CYP2C9 transcripts were either not detected or at a low detection level. To determine whether the encoded enzymes have the potential to activate aflatoxin B1 (AFB1), we used artificial intelligence (AI)-based structural modeling along with molecular docking. AI modeling suggested that CYP3A enzymes can position AFB1 in an orientation compatible with the formation of the reactive intermediate, and CYP3A4 showed the most favorable predicted interaction (docking score: −16.3 kcal/mol). To demonstrate AFB1 bioactivation, we exposed VERO-E6 cells to 200 nmol/L AFB1. After 10 days, we observed about 40% cell death. Liquid chromatography–tandem mass spectroscopy (LC–MS/MS) analysis confirmed the presence of AFB1-derived DNA adducts, indicating that metabolic activation occurred in these cells. These findings support the presence of CYP-dependent AFB1 bioactivation in VERO-E6 cells. Thus, combining computational and experimental approaches elucidates xenobiotic metabolism in cells where biochemical data are limited. Full article

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection causes an immune-mediated neurological syndrome, which persists long after infection. Mechanisms for SARS-CoV-2-associated neurological complications are multifactorial, with an increased risk of drug abuse such as methamphetamine (meth). SARS-CoV-2 infection and its viral proteins play pivotal roles in coronavirus disease 2019 (COVID-19)-associated neuroinflammation, which can lead to long COVID. We hypothesize that meth augments activation of the microglial NOD-, leucine-rich repeat, and pyrin domain-containing protein 3 (NLRP3) inflammasome by the SARS-CoV-2 nucleocapsid (N) protein, resulting in neuroinflammation. To test this hypothesis, we investigated the effect of N-protein and meth on NLRP3 inflammasome activation in primary rat microglial cultures using enzyme-linked immunosorbent assay (ELISA), Reverse Transcription quantitative Polymerase Chain reaction (RT-qPCR), western blot (WB), and immunofluorescence assay (IFA). Our results showed that meth augmented N-protein-induced microglial activation, as evidenced by increased ionized calcium-binding adapter 1 (Iba-1) expression. The addition of meth to the microglial cultures treated with N-protein increased proinflammatory cytokine production. Meth augmentation of N-protein-induced neuroinflammation was further supported by increased inducible nitric oxide synthase (iNOS)-mediated nitric oxide (NO) production. The effects of meth on N-protein-associated inflammatory responses were significantly attenuated by MCC950, a specific NLRP3 inhibitor. Moreover, meth-associated NLRP3 activation was either blocked by the opioid sigma1-receptor (σ1_-R) inhibitor BD1047 or by σ1R siRNA knockdown. Taken together, these results demonstrated that meth augmented SARS-CoV-2 N-protein-induced neuroinflammation via microglial σ1_-R and the NLRP3 inflammasome, which may underlie the pathogenesis of neurological manifestations in COVID-19, such as long COVID with meth abuse. These results may also underscore the impact of drug abuse on long COVID and provide targets for the development of therapeutic strategies to control the neurological outcomes of long COVID. Full article

Mentha pulegium L. is reported to contain phytochemicals known to bind to estrogen receptors and modulate estrogenic effects. A hydroethanolic leaf extract of Mentha pulegium L. (MPE) was prepared, and its effects on uterine and ovarian tissues in immature rats were investigated, focusing on transcriptional endpoint-related estrogenic activity. Female Sprague Dawley rats were treated with varying doses of MPE alone or in combination with estradiol for seven days. Gene expression analysis was performed using reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) to evaluate the effect of MPE on estrogen-responsive biomarkers: Calbindin-D9k (CaBP-9k), Progesterone receptor (Pgr), Trefoil factor 1 (pS2), Intestinal calcium-binding protein integral (Icabp), Integral membrane-associated protein-1 (Itmap1) and Complement component 3 (CC3) genes. In ovarian tissues, MPE treatment decreased CaBP-9k and Icapb expression, with CC3 showing significant decreases in the 200 mg/kg group. Treatments with MPE and estradiol significantly reduced the expression of all estrogen-responsive genes compared to estradiol treatment. In uterine tissues, 1000 mg/kg MPE increased CaBP-9k and pS2 expression significantly but decreased Icapb, CC3, pS2, and Itmap across all treatment groups significantly. Combined estradiol treatment with MPE (500 and 1000) mg/kg showed significantly low CaBP-9k and CC3 expressions. Increased expression of Icapb, Itmap, and pS2 was observed when combined estradiol treatments with MPE (500 and 1000) mg/kg were compared to estradiol treatment. MPE influenced the expression of specific genes in the uterus and ovaries and thus may exhibit endocrine-modulatory activity by multiple mechanisms of action, highlighting its potential complexity in modulating estrogenic responses. Full article

The coronavirus disease 2019 (COVID-19) pandemic highlighted the critical need for rapid, accessible, and accurate diagnostic tools to support timely clinical decision-making, outbreak control, and public health surveillance. Point-of-care testing (POCT) has emerged as an essential component of decentralized healthcare by enabling diagnostic testing outside conventional laboratory settings. This review was based on a structured literature search of PubMed, Scopus, and Web of Science databases covering studies published between January 2020 and January 2026. The review evaluates current advances in COVID-19 POCT technologies, including molecular assays, antigen-based tests, antibody-based assays, biosensor platforms, and artificial intelligence (AI)-assisted diagnostic approaches. Molecular POCT methods, including rapid reverse transcription polymerase chain reaction (RT-PCR), loop-mediated isothermal amplification (LAMP), and CRISPR-based technologies, provide high analytical sensitivity and specificity and are increasingly suitable for decentralized diagnostic applications. Antigen-based assays offer rapid and cost-effective screening solutions, although diagnostic performance may vary depending on viral load, symptom onset, and circulating variants. Antibody-based POCT remains valuable for seroprevalence studies, retrospective diagnosis, and immune-response monitoring rather than acute infection detection. Emerging biosensor technologies and AI-enabled diagnostic systems demonstrate promising analytical capabilities and operational advantages; however, many remain at the prototype or early-validation stage and require further clinical evaluation before widespread implementation. The findings indicate that no single POCT modality is optimal for all clinical scenarios. Instead, molecular, antigen, antibody, biosensor, and AI-assisted approaches provide complementary strengths that support different diagnostic and public health objectives. Continued advances in assay design, digital connectivity, multiplex testing, and variant-resilient detection strategies are expected to further enhance the role of POCT in COVID-19 management and future infectious disease preparedness. Full article