Search Results (42,461)

Accurate detection of Campylobacter in chicken liver is hindered by strong matrix inhibition. This study evaluated sample-processing strategies to improve droplet digital PCR (ddPCR) quantification of Campylobacter coli and Campylobacter jejuni in chicken liver. Mechanical homogenization (Stomacher) and enzymatic/mechanical dissociation (gentleMACS), with and without 8 μm filtration, were compared. Particle-size analysis showed that filtration, especially following gentleMACS treatment, produced smaller, more uniform particles and reduced variability. Percent-degradation assays confirmed that gentleMACS achieved substantially greater tissue disruption than Stomacher homogenization. The multiplex ddPCR assay, which simultaneously targets C. coli and C. jejuni, produced droplet counts comparable to single-target reactions, indicating minimal interference between targets under the conditions tested. In inoculated liver samples, gentleMACS processing yielded droplet counts similar to those obtained from pure cultures, whereas unprocessed liver caused severe matrix interference and inconsistent quantification. Furthermore, gentleMACS-treated samples exhibited strong log-to-log linearity for quantifying C. coli and C. jejuni, enabling detection near 1 genome copy equivalent per reaction. Overall, the results indicate that enzymatic/mechanical dissociation combined with fine-pore filtration improves ddPCR detection of Campylobacter species in chicken liver. Full article

Acute gastroenteritis is a common condition with a wide and often indistinguishable etiology, requiring laboratory support for an accurate diagnosis. Classical diagnostic methods, including stool culture and antigen-based tests, are limited by restricted pathogen coverage and variable sensitivity. In the present study, 548 stool samples from patients with suspected gastroenteritis were tested using the QIAstat-Dx Gastrointestinal Panel 2 and compared with stool culture and rotavirus/adenovirus antigen tests. The molecular panel showed a positivity rate of 50.4%, consistently higher than stool culture (12.6%) and antigen assays (12.3% for rotavirus and 4.4% for adenovirus). The most frequently detected pathogens included enteropathogenic Escherichia coli (15.3%), Campylobacter spp. (12.0%), and enteroaggregative E. coli (10.2%). Agreement between methods was good for bacterial pathogens but low for viral targets. Discordant viral results were often associated with low antigen cut-off index values and multiple pathogen detections by the molecular panel, suggesting potential limitations of antigen-based assays. Overall, the QIAstat-Dx Gastrointestinal Panel 2 improves pathogen detection and provides rapid, comprehensive diagnostic information, while a combined approach with conventional methods may represent the most appropriate strategy for optimizing patient management. Full article

Metastasis remains the leading cause of mortality in breast cancer and is closely linked to epithelial–mesenchymal transition (EMT) and tumor microenvironment (TME)-associated processes. Diosmetin (DT), the active metabolite of diosmin, a widely used venoactive drug, has emerged as a potential anticancer agent. Building on our previous findings demonstrating that DT enhances doxorubicin efficacy, this study investigated its effects on tumor cell plasticity and stromal activation-associated responses. EMT was induced in MCF-7 cells, while a stromal model was established by TGF-β-mediated activation of BJ fibroblasts toward a cancer-associated fibroblast (CAF)-like phenotype. Additionally, doxorubicin-induced senescence was generated in fibroblasts. Migration assays and quantitative real-time PCR were used to assess functional and transcriptional changes. EMT induction resulted in decreased CDH1 expression and increased levels of VIM, MMP2, MMP9, IL-6, and HIF-1A, accompanied by enhanced migratory activity. DT attenuated TGF-β-induced CAF-like activation, as reflected by reduced expression of ACTA2, HGF, MMP2, MMP9, and IL6, and modulated hyaluronan turnover-related genes. Moreover, DT partially alleviated selected senescence-associated features in doxorubicin-treated fibroblasts. Collectively, these findings indicate that DT modulates EMT-associated plasticity and stromal activation-related responses in parallel in vitro models. Given its origin as a metabolite of a clinically used compound and its previously demonstrated chemosensitizing properties, DT may warrant further investigation as a potential adjunctive agent to modulate tumor- and stromal-associated processes in breast cancer. Full article

Potato (Solanum tuberosum L.) is the world’s fourth-largest staple crop. Alkaline salt stress is a major abiotic stress factor that severely limits the growth, yield, and quality of potatoes; however, little is known about the molecular basis of potatoes’ response to alkaline salt stress or the stress-alleviation mechanism mediated by 24-epibrassinoside. In this study, we conducted a genome-wide identification of the potato miR482 family and analyzed its response patterns under alkaline salt stress and 24-epibrassinoside-mediated stress relief. We identified a total of 9 mature stu-miR482 sequences and 5 precursor sequences; all precursors form typical stable hairpin structures and exhibit high evolutionary conservation among Solanaceae plants. Promoter analysis revealed multiple cis-acting elements in the promoter region associated with light signaling, plant hormones, and stress signaling. A total of 64 potential target genes were predicted, encompassing transcription factors, disease resistance, and signal transduction-related genes, forming a complex regulatory network. Phenotypic analysis confirmed that EBR significantly alleviates the growth inhibition in potatoes induced by alkaline salt stress. qRT-PCR analysis indicated that stu-miR482a-5p is the primary stress-responsive member in leaves; stu-miR482d-3p/5p exhibited the strongest regulatory response to EBR in roots; in potato stolons, all members of the miR482 family were significantly upregulated under alkaline salt stress, with stu-miR482d-5p showing extremely significant upregulation across all treatment groups. In summary, this study represents the first systematic characterization of the potato miR482 family, revealing its tissue differential functions in alkaline salt stress and EBR-mediated stress relief. Full article

Background: The Taiwanese loach (Paramisgurnus dabryanus ssp. Taiwan, Dabry de Thiersant, 1872.) is an economically important aquaculture species in East Asia, and its body color directly affects its ornamental and market value. Our research group recently discovered a golden-red mutant, named “Gan Hong No. 1” (MR), within a wild-type (WT) population. During embryogenesis, MR individuals exhibit almost no melanophore deposition, and after hatching, xanthophores and erythrophores appear sequentially, suggesting that the body color variation likely originates from alterations in the gene regulatory network during early development. Objective: To systematically compare the transcriptomes of WT and MR-Taiwanese loach during early development, to identify the key regulatory pathways underlying red body color formation from a temporal perspective, to test whether the classical melanin synthesis pathway is impaired, and to provide a theoretical basis for selective breeding of body color traits. Methods: High-throughput transcriptome sequencing was performed on eight early developmental stages (0, 5, 10, 15, 20, 23, 28 and 43 h post-fertilization) of both loach types. Differential expression analysis, time-series trend analysis, and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment were used to systematically characterize gene expression dynamics. Transcriptomic data validation was performed using real-time PCR. Results: In MR, the core transcription factor mitfa was significantly downregulated, whereas the expression of melanin synthesis genes such as kita and dct showed no significant difference, indicating that the impairment of melanogenesis is caused by mitfa downregulation. Trend analysis and pathway enrichment revealed that in MR embryos, pathways related to oxidative stress, unsaturated fatty acid biosynthesis, C-type lectin receptor signaling, p53 signaling, and apoptosis were significantly activated, while the thyroid hormone synthesis pathway was markedly upregulated. In WT, these pathways showed the opposite trend. qRT-PCR results were consistent with the transcriptome data. Conclusions: This study demonstrates that downregulation of mitfa serves as the initial trigger for red body color variation in the Taiwanese loach. This mutation impedes melanin synthesis and concurrently activates a coordinated regulatory network involving oxidative stress, immune inflammation, and thyroid hormone signaling. Accumulation of unsaturated fatty acids alleviates oxidative damage and supports carotenoid deposition, while immune signals eliminate aberrant melanocytes and promote compensatory generation of red and yellow chromatophores. The upregulated thyroid hormone further fine-tunes pigment cell differentiation. For the first time in a cobitid species, this study elucidates the mitfa-mediated, multi-pathway synergistic molecular mechanism driving the transition from melanin-based to carotenoid/pteridine-based red coloration in fish, thereby providing a theoretical reference for molecular breeding of body color in aquaculture. Full article

Background/Objectives: Leukocyte telomere length (TL) is a marker of biological aging associated with cardiovascular disease, chronic kidney disease, and malignancy in the general population. Its long-term prognostic significance in systemic lupus erythematosus (SLE) remains unclear. We aimed to evaluate the association between baseline TL and long-term clinical outcomes in patients with SLE. Methods: Prospective cohort study including 97 Caucasian women with SLE. Relative TL was measured in whole blood using quantitative polymerase chain reaction (qPCR) at baseline. A control group of 50 healthy Caucasian women from the same geographical region was included for comparison. Patients were followed for a mean of 9.7 ± 2.8 years. Outcomes included thrombotic cardiovascular events, damage accrual, incident malignancy, and chronic kidney disease. Associations were assessed using multivariable regression models adjusted for potential confounders. Results: Mean age was 51.6 ± 13.8 years and mean relative TL was 4.3 ± 1.0. Relative TL was inversely associated with age (β = −0.20, p = 0.048) and was shorter in patients with hematological manifestations (p = 0.038). No differences in relative TL were observed between SLE patients and controls. Relative TL was not associated with disease activity, cumulative damage, cardiovascular risk factors, vitamin D levels, or subclinical atherosclerosis. During follow-up, 13.4% of patients experienced cardiovascular events, 10.3% developed malignancy, and 11.3% developed chronic kidney disease. Relative TL was initially associated with long-term damage accrual, glomerular filtration rate and cardiovascular events; however, after adjustment for age, only the association with glomerular filtration rate remained at the limit of statistical significance (p = 0.05). Conclusions: In this prospective cohort, relative TL was primarily associated with aging, hematological manifestations, and glomerular filtration rate, but not with disease activity or most long-term clinical outcomes. These findings suggest that TL reflects biological aging rather than disease-specific processes and has limited utility as a prognostic biomarker in SLE. Full article

Splicing factor 3b1 (SF3B1), a component of U2 small nuclear ribonucleoprotein (U2 snRNP), has been known for its essential roles in pre-mRNA splicing and alternative splicing. Here we show that knocking down (KD) of SF3B1 broadly induced a significant reduction in mRNA expression in the genome. One of the genes whose expression is reduced by SF3B1 KD is methyl-transferase-like 3 (METTL3), a writer of N6-methyladenosine (m⁶A). We demonstrate that expression of both METTL3 mRNA and protein is affected by SF3B1 KD, which further decreases the m⁶A RNA expression level. m⁶A-seq indicates that SF3B1 KD affects m⁶A distribution within multiple genes in the genome. In addition, a high proportion of hypo-methylation events by SF3B1 KD (~70%) are overlapped in METTL3 KD cells, and a conserved m⁶A motif is observed in the hypo-methylated regions as in SF3B1 KD cells, suggesting the m⁶A decrease by SF3B1 is a direct effect of the reduced METTL3 expression. Furthermore, RT-qPCR using unlabeled RNA and 5-Bromouridine (BrU)-labeled nascent RNA and actinomycin D treatment demonstrates that transcription of METTL3 is significantly reduced but the mRNA decay rate is not altered, suggesting that METTL3 expression is altered at the transcription level. We further show that SF3B1 interacts with RNA polymerase (Pol) II in the RNA independent manner, further indicating the involvement of SF3B1 in transcription. Lastly, we demonstrate that the transcription inactive H3K27me3 on the METTL3 promoter was significantly increased whereas transcription active H3K4me3 was not changed by SF3B1 KD. Taken together, we conclude that reduced SF3B1 expression suppresses the transcription of METTL3 and inhibits m⁶A RNA expression. Full article

Objectives: Noninvasive prenatal testing relies on the analysis of total cell-free DNA (cfDNA) in maternal plasma, where fetal-derived DNA constitutes only a minor fraction. This study aimed to preliminarily compare a modified TPY cfDNA extraction protocol with two commercial extraction kits for the downstream detection of Y-chromosome-specific markers in pregnancies carrying male fetuses. Methods: Plasma samples were obtained from 52 singleton pregnancies between 10 and 30 weeks of gestation with male fetal sex confirmed by ultrasonography. Total cfDNA was extracted from aliquots of the same maternal plasma samples using the modified TPY protocol, the QIAamp DSP Virus Kit, and the MagMAX™ Cell-Free DNA Isolation Kit. Quantitative real-time PCR was performed for the Y-chromosome-specific markers SRY and DYS14. At the same time, GLO was used as a reference marker to reflect the total cfDNA background. Extraction performance was assessed primarily using total cfDNA concentration and Ct values obtained from amplification of fetal-specific Y-chromosome markers. Results: Total cfDNA concentrations varied among the extraction methods, with the commercial kits yielding higher total cfDNA concentrations than the modified TPY protocol. In contrast, the TPY protocol yielded slightly lower mean Ct values for SRY and DYS14 than the commercial kits. SRY and DYS14 amplification was detected in 90.4% and 94.2% of samples, respectively. However, these Ct differences should be interpreted cautiously because fetal fraction, maternal DNA contamination, extraction recovery, and fragment size distribution were not directly measured. Conclusions: The modified TPY protocol showed preliminary technical feasibility for extracting total cfDNA from maternal plasma and enabling downstream amplification of Y-chromosome-specific markers in male pregnancies. Nevertheless, the observed lower Ct values do not establish selective fetal DNA enrichment, reduced maternal DNA contamination, or clinical superiority over commercial methods. Further analytical validation using standardized fetal fraction measurement, recovery efficiency testing, fragment size analysis, fetal-to-maternal DNA ratio assessment, and larger cohorts including both male and female pregnancies is required before broader clinical applicability can be determined. Full article

Bioprospecting for extremozymes from unique ecological niches is crucial for developing robust biocatalysts for green chemistry. Here, we report the de novo hybrid genome assembly of Penicillium crustosum CTM10622, isolated from the humid montane forest of El Feïdja National Park, Tunisia. Using Illumina NextSeq™ 500 and Nanopore PromethION 2 Solo, a highly contiguous 31.38 Mb assembly (N50 = 1.94 Mb; 98.3% BUSCOs) was achieved. This robust genomic foundation enabled the identification of an extensive hydrolase repertoire, leading to the discovery of a novel alkaline serine lipase, PCLIP, subsequently heterologously expressed in Pichia pastoris. Recombinant rPCLIP exhibited a high specific activity (15,000 U/mg at pH 10, 65 °C) and exceptional thermostability, with half-lives of 14 and 8 h at 80 and 90 °C, respectively. The enzyme’s identity as a serine lipase was confirmed by its complete inhibition by Orlistat or tetrahydrolipstatin (THL) (51 µM), PMSF (5 mM), and diisopropylfluorophosphate (DIFP) (2 mM). To determine its substrate specificity, advanced computational approaches, including convolutional neural network-based docking and explicitly solvated molecular dynamics, were employed to compare rPCLIP with its homologue PCrL, a recombinant serine alkaline lipase from Penicillium crustosum Thom P22. While rPCLIP showed optimal experimental activity toward short-chain glyceryl tributyrate, simulations revealed that long-chain trioctanoin acts as a ‘thermodynamic trap’ due to over-stabilization. Conversely, the rigid rPCrL favors tricaprylin, driven by a ‘hydrophobic engine’ effect where the solvated environment forces chain burial with minimal entropic penalty. The findings demonstrate that rPCLIP specificity is driven by a delicate interplay of geometric complementarity, Van der Waals enthalpy, and conformational entropy. Full article

The purpose of this study was to investigate the adverse effects of 1.5 μg·L⁻¹ environmentally relevant chlorantraniliprole (CAP) on oxidase biomarkers (juvenile, 2.5 g) for 2, 4, and 8 h and transcriptomic response (adult, 254.8 g) for 96 and 192 h in American shad Alosa sapidissima (Wilson, 1981). American shad is sensitive to pollutants and has become an important economic fish in China, especially for recirculating the aquaculture system and photovoltaic farming. For juvenile shad under short-time CAP exposure, acid phosphatase (ACP) and aryl hydrocarbon receptase (AHR) at the protein level significantly increased at 2 h, and for longer-time exposure, alkaline phosphatase (AKP), polyphenol oxidase enzyme (PPO), and tumor necrosis factor alpha (TNFα) at the protein level significantly decreased; ryanodine receptase (RYR) at the protein level was significantly increased at 8 h. Interestingly, malondialdehyde (MDA) contents, biomarkers of oxidative stress, were significantly decreased for depletion at 2 h and 4 h, while they increased for eliminating free radicals at 8 h via longer-time CAP exposure duration. With the same CAP exposure for adult shad, the number of congested and dilated sinuses of the liver changed, with fine granular brown pigmentation and vacuolization of hepatocytes at 96 h, while the sinuses and central veins were dilated and edematous degeneration occurred at 192 h for longer-time exposure. The detected enzymatic activities, except for adenosine 5′-monophosphate (AMP)-activated protein kinase (AMPK), significantly decreased, and MDA contents significantly increased in adult shad at 96 and 192 h. Ribosome, proteasome, spliceosome, protein processing in endoplasmic reticulum, oxidative phosphorylation, glycerophospholipid metabolism, biosynthesis of amino acids, ferroptosis, peroxisome, apoptosis, necroptosis, and mTOR signaling pathways were the most significantly enriched pathways. For qPCR verification, the genes ppa2, pla1a, psmb13a, pkz and stat1b were significantly upregulated, while hspa8b, capn2, tram2, asns, bcl2l1, diablo, and prkcb were downregulated in adult shad. The results reveal elevated oxidative stress causing time-dependent hepatic damage via 1.5 μg·L⁻¹ CAP exposure both in juvenile and adult shad. Full article

The global rise in multidrug-resistant (MDR) fungal pathogens has positioned Candida auris and Candida glabrata as major threats to public health. In recent years, these pathogens have increasingly been reported beyond traditional hospital settings, including neonatal intensive care units, long-term care facilities, oncology wards, and post-pandemic critical care environments. International surveillance bodies, including the Centers for Disease Control and Prevention (CDC), European Centre for Disease Prevention and Control (ECDC), World Health Organization (WHO), and regional monitoring networks, have documented escalating antifungal resistance, complex outbreak dynamics, and persistent gaps in infection control implementation. C. auris has emerged as a major etiological agent of healthcare-associated outbreaks, particularly in intensive care and neonatal units. Surveillance data indicate that a high proportion of C. auris isolates exhibit resistance to azoles, often exceeding 80% in some regions, while echinocandin resistance remains variable. Resistance patterns have evolved from predominantly azole resistance to broader multidrug-resistant phenotypes, including treatment-emergent echinocandin resistance. Six genetically distinct clades (I–VI) have been identified, with Clades I, III, and IV associated with large-scale outbreaks, whereas available data suggests that Clades II, V, and VI are more geographically restricted, although evidence for the recently described clades remains limited. C. glabrata is increasingly recognized as a major cause of invasive candidiasis, with rising resistance reported across multiple regions. While reduced azole susceptibility was historically predominant, emerging evidence highlights rising dual azole–echinocandin resistance, adaptive microevolution during antifungal therapy, and biofilm-associated tolerance mechanisms. Despite these advances, significant gaps persist in global resistance surveillance and in the mechanistic understanding of virulence and antifungal adaptation. Current mitigation strategies include antifungal stewardship programs, expanded resistance testing, and strengthened surveillance systems. Advances in rapid diagnostic technologies such as matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) mass spectrometry, polymerase chain reaction (PCR)-based assays, and genomic surveillance have improved pathogen identification and outbreak detection, although accessibility remains limited in resource-constrained settings. This review examines emerging epidemiological, genomic, and antifungal resistance trends in C. auris and C. glabrata and highlights key priorities for improving diagnosis, surveillance, stewardship, and management of multidrug-resistant Candida infections. Full article

Plasma membrane intrinsic proteins (PIPs), a subfamily of aquaporins (AQPs), play critical roles in various physiological processes in plants, including the transport of water and CO₂, regulation of stomatal movement, absorption of neutral molecules and nutrients, and H₂O₂ signaling. Nevertheless, the functions of PIP aquaporins in adventitious root formation in trees are still poorly understood. PtrPIP2:4 is specifically expressed in roots, and PtrPIP2:4 fused with GFP localizes to the plasma membrane. Overexpression of PtrPIP2:4 significantly accelerated adventitious root induction in poplar. Stem cuttings from overexpression lines exhibited more rapid rooting compared to wild-type (WT) plants, although the total number of adventitious roots did not differ significantly. Additionally, the number of lateral roots was markedly increased in PtrPIP2:4 overexpression lines. Comparative transcriptome analysis identified 4204 differentially expressed genes (DEGs) between WT and PtrPIP2:4 overexpression plants. Transcriptomic analysis revealed that genes associated with auxin-related and flavonoid biosynthesis were significantly enriched. RT-qPCR results showed that the transcription levels of nine auxin-related genes (i.e., PtrARF, PtrIAA, PtrGH3 and PtrPIN) were significantly upregulated, while the transcription levels of five flavonoid synthesis genes (i.e., PtrDFR, PtrANS, PtrANR and PtrLAR) were also significantly upregulated. Previous studies have implicated these genes in adventitious root formation. Collectively, these findings reveal that PtrPIP2:4 accelerates adventitious root emergence, promotes adventitious root elongation, and increases lateral root number while the total number of adventitious roots exhibited no significant difference in poplar, suggesting its potential utility in improving tree propagation and breeding strategies. Full article

Peacekeeping operations in sub-Saharan Africa continue to be impacted by malaria both in-country and among returning service members. The Royal Thai Army (RTA) deploys an engineering company to Juba and Rumbek, South Sudan to conduct peacekeeping operations as part of the UN Mission in South Sudan (UNMISS). Each deployment is approximately 12 months long. The unit is given doxycycline one week before travel before switching to the UN-provided mefloquine during deployment and for four weeks after returning. The RTA routinely conducts post-deployment screening for malaria by microscopy and PCR for units returning from UNMISS. High rates of prophylaxis failure were observed from both during-mission and post-deployment screening cases, with cumulative malaria attack rates of 11.4% (31 cases out of 271 personnel), 18.2% (49 cases out of 270 personnel), and 23.1% (63 cases out of 273 personnel) for 2023, 2024, and 2025, respectively, with 98% of cases being due to P. falciparum. Furthermore, post-deployment screening revealed high rates of sub-microscopic and sub-clinical parasitemia with 40% of all malaria cases being identified as asymptomatic during post-deployment screening, and 61% of those asymptomatic cases being detected by PCR only. While factors underlying the high prophylaxis failure rate, as well as the high rate of sub-microscopic and sub-clinical parasitemia are unclear, these findings highlight the limitations of relying on clinical symptoms or microscopy for detecting malaria in military units returning from endemic regions and underscore the importance of unit-wide post-deployment molecular screening. Full article

The Yangtze sturgeon (Acipenser dabryanus), a critically endangered living fossil whose wild populations are now extinct, faces new challenges to survival in captive breeding. Among these, the emergence of albino and gray color morphs raise fundamental questions about the molecular basis and physiological consequences of pigmentation loss. Here, we integrated histological, transcriptomic, and quantitative PCR to investigate pigmentation variation and associated immune alterations in this species. Histology revealed a complete absence of melanin in albino individuals and marked reduction in gray morphs. Transcriptomic profiling across the three color morphs uncovered a broad downregulation of core melanogenic genes, including PMEL, TYR, TYRP1, DCT, SLC45A2, OCA2, MREG, and MLPH, indicating impaired melanosome formation, melanin synthesis, and intracellular transport. Notably, pigmentation loss coincided with systematic changes in the expression of immune-related genes: phagosome pathway genes (e.g., C3, MHC I/II, TAP2) were downregulated, while pro-inflammatory mediators (e.g., IL-8, IL-17, CXCL10) were upregulated, suggesting a transcriptional pattern correlated with reduced expression of pathogen defense-related genes and increased genes associated with inflammation mediators. These findings reveal a mechanistic correlation between melanin deficiency and immune dysfunction in a basal vertebrate lineage, offering the first molecular evidence of an association between albinism and altered immune-related gene expression in sturgeons and highlighting its implications for conservation and captive management. Full article

To improve clinical decision-making about Carbapenem-resistant Gram-negative bacteria (CR-GNB) infections and halt the spread of resistant microbes, quicker and less expensive diagnostic techniques are required. Thus, the purpose of this study was to thoroughly evaluate the diagnostic efficiency (sensitivity, specificity, and concordance) of direct-from-specimen multiplex lateral flow immunoassay (LFIA) across diverse raw clinical specimens and pathogen types from critically sick patients. A total of 300 non-duplicate samples were tested to detect CR-GNB. Five major Carbapenemase genes were detected directly from the specimen using carbapenem-resistant K.N.I.V.O. detection K-Set and from culture using culture-enhanced multiplex PCR. Turnaround time (TAT) of each method was calculated. The direct LFIA revealed 100% specificity for NDM, KPC, and IMP enzymes in all tested clinical matrices (blood, urine, and respiratory samples). The study demonstrated 100% sensitivity and specificity with perfect categorical agreement (κ = 1.000) for the blaKPC in the Klebsiella pneumoniae and for blaOXA-48 and blaIMP in the Acinetobacter baumannii; however, sensitivity of blaVIM was significantly diminished across all isolates and samples. TAT decreased significantly (p < 0.001) from 30 to 70 h to about 50 min. The tested direct LFIA facilitates the prompt enhancement of lifesaving tailored antibiotic treatment for severe illnesses. Full article