Search Results (628)

Tuberculosis is still one of the leading infectious causes of morbidity and mortality worldwide. Rapid diagnosis is essential for effective treatment and control of tuberculosis transmission. In recent years, nucleic acid amplification tests (NAATs), such as GeneXpert MTB/RIF, BD MAX™, Xpert MTB/RIF-Ultra, have significantly improved tuberculosis diagnostics. However, they mainly require expensive and advanced equipment. The aim of our study was to assess the usefulness of the novel RITA MTBC assay in this diagnostic context. A total of 61 clinical specimens were tested using the RITA MTBC assay in comparison with established molecular diagnostic platforms (GeneXpert and BD MAX™), used as molecular reference assays. Culture and microscopy were performed as part of initial clinical assessment, but the comparative analysis focused on molecular assays to provide a relevant evaluation of diagnostic performance. Among 31 samples previously identified as positive for M. tuberculosis DNA, the assay correctly detected 30 (LOT HPA01/20230601) and 29 (LOT HPA01/20230602). Of 30 negative samples, 28 and 30 were confirmed negative for the respective lots. These results correspond to an average sensitivity of 95% and an average specificity of 97%. The kit demonstrated diagnostic performance that meets requirements for molecular testing in tuberculosis, with sensitivity and specificity comparable to established platforms, although further validation on larger sample sets is necessary. Nevertheless, its excellent specificity, rapid turnaround time, and operational simplicity, make it especially well-suited for decentralized or resource-limited settings. These findings underscore the potential of RITA MTBC as a valuable diagnostic tool in both routine clinical settings and in populations with limited access to healthcare. Full article

Aim: to study the level of immunoglobulin FLC in patients with myocarditis in comparison with non-inflammatory heart diseases, and FLC’s correlation with the severity of CHF. Methods: Ninety-nine patients (41 women, 59.6 ± 14.6 y.o.) were included in the study: 50 patients with myocarditis [confirmed by myocardial biopsy (n = 20) and/or cardiac MRI]; 49 patients with non-inflammatory heart disease. CHF was diagnosed in 66% and 65% of patients, respectively. The levels of FLC were determined using the ‘Cloneus S-FLC-K TIA Kit’ and ‘Cloneus S-FLC-L TIA Kit’ reagents. Results: Elevated FLC levels were found in 56% of patients with myocarditis and in 67% of comparison group patients (p > 0.05). Mean FLC kappa levels were 13.4 [11.7; 16.7] and 16.0 [11.3; 23.7] mg/L, FLC lambda 22.7 [16.7; 32.4] and 24.7 [18.1; 39.1] mg/L, FLC kappa/lambda ratio 0.62 [0.50; 0.73] and 0.65 [0.56; 0.76] in myocarditis and comparison groups, respectively; there were no significant differences between groups. Both groups showed correlations of FLC levels with levels of CRP and leukocytes, as well as with glomerular filtration rate, CHF NYHA class, and left ventricular ejection fraction. Only in patients with myocarditis did we observe a significant correlation between both kappa and lambda FLC and NT-proBNP (r = 0.528, p = 0.004, and r = 0.756, p < 0.001) and high-sensitivity troponin (r = 0.829, p = 0.042) levels. Conclusions: Increased FLC level may be considered an important pathogenetic link reflecting both specific mechanisms of myocarditis and severity of CHF. The determination of FLC can be used as an additional diagnostic marker, as well as one predictor of the decompensated course of myocarditis. Full article

Studies in human medicine have demonstrated that rotavirus infection can also affect extraintestinal sites due to its systemic effects. However, in veterinary medicine, the injury caused by rotavirus diarrhea is limited to the intestines, and its effects on various systemic structures remain poorly understood. In this observational case–control study, we aimed to determine the effects of HSP-27, Caspase-3, IL-2, γ-H2AX, HMGB-1, SP-D, and GDH (or GLDH) on the pathogenesis of rotavirus infection by using biomarkers for diagnostic purposes in lung and liver injury in neonate diarrheic calves naturally infected with rotavirus, both alive and post-mortem. Fifty-two Simmental calves (1–28 days old) of both sexes, 40 infected with rotavirus and 12 healthy, were studied. Twenty-eight out of 40 survived, while the remainder underwent necropsy for histopathological and immunopathological (HSP-27, Caspase-3, IL-2, γ-H2AX) examination of the lungs and livers. Lung and liver-specific serum E-selectin, glutamate dehydrogenase, surfactant protein-D, and high mobility group box-1 were analyzed by a bovine-specific ELISA kit (Shanghai Coon Koon Biotech Co., Ltd., China). Histopathological and immunohistochemical analyses confirmed lung and liver injury in naturally infected calves. HMGB-1, SP-D, and GDH concentrations were significantly higher in naturally infected calves than in the control group (p < 0.001, p < 0.001, and p < 0.05, respectively), showing an excellent diagnostic predictive capacity for lung and liver injury. Also, IL-2, HSP-27, CASP-3, and γ-H2AX were significantly expressed in the lungs (p < 0.001, p < 0.001, p < 0.001, and p < 0.05, respectively) and liver (p < 0.001, p < 0.001, p < 0.01, and p < 0.01, respectively). All these observations led us to hypothesize that oxidative stress, apoptosis, and DNA damage may underlie the pathogenesis of this condition. Nevertheless, further studies on large populations of rotavirus-infected calves are needed to confirm the data reported in the current study. Full article

Proteoglycans and their fragments have potential as diagnostic or theragnostic biomarkers to identify diseases characterized by dysregulated extracellular matrix remodeling, such as juvenile idiopathic arthritis (JIA). Therefore, our study aimed to evaluate the diagnostic utility of plasma proteoglycan profiles, namely, aggrecan, decorin, and biglycan, released from osteoarticular structures into the blood of children with juvenile idiopathic arthritis. These profiles are potential biomarkers of tissue destruction and/or indicators of the efficacy of therapy with the biologic agent etanercept (ETA). This study was conducted on 263 blood samples collected from 25 healthy children and 34 children at various stages of juvenile idiopathic arthritis disease: immediately after diagnosis, following treatment with disease-modifying antirheumatic drugs (DMARD) (methotrexate, sulfasalazine, and prednisone), and during 3, 6, 12, 18, and 24 months of therapy with etanercept. Quantitative levels of aggrecan, biglycan, and decorin were measured using ELISA kits. In children with JIA, plasma aggrecan levels were elevated at diagnosis, decreased after ineffective DMARD therapy, and increased again at 24 months of etanercept treatment despite clinical improvement. By contrast, biglycan levels were similar to those in healthy controls but decreased during etanercept therapy. Decorin levels were initially high in untreated and DMARD-treated patients but returned to normal after 24 months of biologic treatment. After considering these findings and the ROC analysis, we conclude that decorin appears to be a promising biomarker for diagnosing and monitoring etanercept therapy in JIA, and biglycan is a useful biochemical marker for assessing the effectiveness of ETA treatment. Full article

This study documents an outbreak of Salmonella enterica serovar Enteritidis (SE) on a commercial meat rabbit farm in Italy. Following the observation of increased mortality in kits and severe enteric symptoms across all age groups, SE was first isolated in early March 2024. A diagnostic and epidemiological investigation was subsequently undertaken to characterize the anatomo-histopathological features in deceased rabbits and to identify the source and transmission dynamics of the infection. Between March and December 2024, a total of 1550 rectal swabs from live rabbits, 60 environmental samples, and 168 carcasses were collected and subjected to microbiological analysis. SE-positivity rates ranged from 8.4% to 36.3%, depending on the sample type considered. Co-infections with Pasteurella multocida, Escherichia coli, and Staphylococcus spp. were also detected. Gross and histological lesions in SE-positive rabbits included fibrinonecrotizing enterocolitis, hepatosplenomegaly, and renal damage such as suppurative nephritis and tubulorrhexis. Despite the implementation of enhanced biosecurity protocols, SE re-emerged over time and across different pens. Given the zoonotic potential of SE, the outbreak described underscores the need for rabbit-specific Salmonella control programs to safeguard both animal and public health. Full article

The West Nile Virus (WNV), transmitted by Culex mosquitoes as a major vector, has been reported worldwide. Also, West Nile neuroinvasive disease (WNND) caused by WNV lineage 1a and 2 neuroinvasive infections has been constantly reported with high fatality rates. Nevertheless, there are no treatments and vaccinations, so diagnosis in the early stages is important. Recently, a molecular diagnostic technique using DNA endonuclease-targeted CRISPR trans reporter (DETECTR) with the CRISPR-Cas12a system integrated with isothermal nucleic acid amplification has newly emerged. In this study, we designed a 2-Step WNV DETECTR with reverse transcription–recombinase polymerase amplification (RT-RPA) for rapid and sensitive WNV diagnosis. It successfully detected down to 1.0 × 10² RNA copies for both WNV lineage 1a and 2 with demonstrating similar sensitivity to qRT-PCR without cross-reactivity to other viruses. Additionally, we designed a 1-Step WNV DETECTR, incorporating all processing steps into a single tube, capable of detecting down to 1.0 × 10³ RNA copies for both lineages. Furthermore, we developed a more streamlined method, the 1-Step with Filter WNV DETECTR, which achieved detection limits comparable to the 2-Step method, while reducing the processing time by 5 min. This study also explored the potential of the Punch-it™ NA-Sample Kit as an efficient alternative lysis method by comparing the detection differences across various lysis methods. Through this method, we achieved rapid and simple amplification and detection processes suitable for field diagnostics with high specificity and sufficient sensitivity. Therefore, DETECTR methods presented themselves as promising alternatives to conventional diagnostic tools, potentially overcoming financial and technical constraints in diverse medical settings. Full article

Apart from some information on dengue virus (DENV), there is limited data on the circulation of arboviruses in Burkina Faso. The aim of this study was to investigate antibody prevalence against six arboviruses in four regions of the country to document previous virus exposure. Serum samples collected between August 2018 and December 2022 from people infected with viral hepatitis B and C in Bobo-Dioulasso were used to detect IgG antibodies against DENV, Chikungunya virus (CHIKV), Zika virus (ZIKV), Yellow fever virus (YFV), Rift Valley fever virus (RVFV) and Crimean-Congo hemorrhagic fever virus (CCHFV) using commercial ELISA kits. A total of 1808 serum samples, accompanied by basic epidemiologic data (sex, age and residency) were included in this study. We observed an IgG antibodies seroprevalence of 75.4% for DENV, 30.8% for CHIKV, 2.9% for ZIKV, 1.2% for RVFV, 1.1% for CCHFV and 1.1% for YFV. Age, sex, and place of residence were significantly associated with seropositivity for DENV and age and sex with CHIKV seropositivity. The results suggested widespread circulation of DENV and CHIKV and possible circulation of CCHFV and RVFV in humans in Burkina Faso. The importance of strengthening arbovirus surveillance by including additional arboviruses in the diagnostic panel is emphasized. Full article

Background and Clinical Significance: Tyrosine kinase inhibitors (TKIs) have transformed Philadelphia chromosome-positive chronic myeloid leukemia (Ph+ CML) into a largely manageable chronic disease. However, off-target toxicities are increasingly recognized; rarer complications such as bone marrow edema (BME) remain underreported. BME is a radiological syndrome characterized by excess intramedullary fluid on fat-suppressed T2/STIR magnetic resonance imaging sequences and may progress to irreversible osteochondral damage if unrecognized. We report a case series of TKI-associated BME and propose a practical diagnostic-therapeutic framework. Case Presentation: We describe three patients with Ph+ CML who developed acute, MRI-confirmed BME of the lower limb during TKI therapy. Case 1 developed unilateral then bilateral knee BME, temporally associated first with dasatinib and subsequently with imatinib; symptoms improved after TKI interruption, bisphosphonate therapy, and supportive measures, and did not recur after switching to bosutinib. Case 2 presented with proximal femoral BME during long-term imatinib; imatinib was stopped, intravenous neridronate administered, and bosutinib initiated with clinical recovery and later near-complete radiological resolution. Case 3 experienced multifocal foot and ankle BME during imatinib; symptoms resolved after drug discontinuation and bisphosphonate therapy, and disease control was re-established with bosutinib without recurrence of BME. All patients underwent molecular monitoring and mutational analysis to guide safe therapeutic switching. Discussion: Temporal association across cases and the differential kinase profiles of implicated drugs suggest PDGFR (and to a lesser extent, c-KIT) inhibition as a plausible mechanistic driver of TKI-associated BME. PDGFR-β blockade may impair pericyte-mediated microvascular integrity, increase interstitial fluid extravasation, and alter osteoblast/osteoclast coupling, promoting intramedullary edema. Management combining MRI confirmation, temporary TKI suspension, bone-directed therapy (bisphosphonates, vitamin D/calcium), symptomatic care, and, when required, therapeutic switching to a PDGFR-sparing agent (bosutinib) led to clinical recovery and preservation of leukemia control in our series. Conclusions: BME is an underrecognized, potentially disabling, TKI-related adverse event in CML. Prompt recognition with targeted MRI and a multidisciplinary, stepwise approach that includes temporary TKI adjustment, bone-directed therapy, and consideration of PDGFR-sparing alternatives can mitigate morbidity while maintaining disease control. Prospective studies are needed to define incidence, risk factors, optimal prevention, and management strategies. Full article

Background: Rapid and high-throughput diagnostic methods are essential for controlling the spread of infectious diseases, such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Lateral flow immunoassay (LFIA) strips provide a cost-effective and user-friendly platform for point-of-care testing. However, the sensitivity of conventional LFIA kits is often limited by the performance of their detection probes. This study reports a highly sensitive LFIA strip for detecting the SARS-CoV-2 nucleocapsid (NP) protein using platinum-decorated poly(2-vinylpyridine) nanoparticles (Pt-P2VPs) as probes. Methods: Monoclonal antibodies against SARS-CoV-2 NP were conjugated with Pt-P2VPs and incorporated into LFIA strips. The test line was coated with anti–SARS-CoV-2 NP monoclonal antibody, and the control line with goat anti-mouse IgG. Recombinant proteins, viral strains, and nasopharyngeal swab specimens from patients were used to evaluate assay performance, with reverse transcription polymerase chain reaction (RT-PCR) as the reference standard. Diagnostic accuracy was assessed using nonparametric statistical tests. Results: Pt-P2VP-based LFIA strips enabled sensitive detection of recombinant NP and inactivated SARS-CoV-2, with minimal cross-reactivity. In 200 clinical specimens (100 PCR-negative and 100 PCR-positive), the assay achieved 74% sensitivity and 100% specificity, with strong correlation to viral RNA load. Compared with conventional LFIA kits, Pt-P2VP strips demonstrated superior sensitivity at lower viral loads. Conclusions: Pt-P2VPs represent a promising probe material for enhancing LFIA performance and may facilitate the development of rapid, sensitive, and scalable immunoassays for infectious disease diagnostics in biomedical applications. Full article

Background/Objectives: Cuproptosis, a newly discovered form of programmed cell death, is dependent on the regulation of copper ions. The roles and mechanisms of cuproptosis-related genes (CRGs) in the instability of atherosclerotic plaques are still unclear. Methods: GEO microarray datasets were downloaded to analyze stable and unstable human carotid artery plaques. Differential expression analysis was performed to screen for CRGs from the Molecular Signatures Database (MSigDB). Machine learning was applied to identify key genes and cluster unstable plaque genes. The identified genes were verified by immunohistochemistry (IHC) of human carotid plaque samples, and the effect of ATOX1 on cuproptosis was detected in human umbilical vein endothelial cells (HUVEC). Results: This study identified 27 CRGs differentially expressed between stable and unstable plaques. Five characteristic genes (LC3A, ATP7B, ATOX1, CTR1, and NLRP3) were selected by machine learning. A diagnostic model for unstable plaques was developed based on these genes. The expression of ATOX1 and NLRP3 was increased, while LC3A and ATP7B were decreased in unstable plaques. However, there was no significant change in CTR1. The Cell Counting Kit-8 (CCK-8) assay indicated that inhibiting ATOX1 reduced CuSO₄-induced HUVEC death. Conclusions: CRGs appear to influence atherosclerotic plaque formation. Five key genes (LC3A, ATP7B, ATOX1, CTR1, NLRP3) were identified as being differentially expressed in unstable plaques. Cluster analysis uncovered two subtypes (C1, C2) linked to cuproptosis and immune infiltration in unstable plaques. These genes likely affect atherosclerosis progression by influencing immune cell infiltration, thus impacting plaque stability. Furthermore, the cuproptosis-related gene ATOX1 can regulate CuSO₄-induced HUVEC death. This study contributes to predicting plaque instability and offers potential diagnostic and therapeutic targets. Full article

Background/Objectives: Antimicrobial resistance (AMR) represents a major global health challenge, driving the need for rapid and accurate diagnostic tools. Novel molecular assays, including multiplex PCR and DNA microarray-based systems, have emerged to detect antimicrobial resistance genes (ARGs) alongside bacterial identification. Methods: In this study, we evaluated the performance of the HybriSpot12 PCR AUTO (HS12a) system and the MDR Direct Flow Chip (MDR-FC) Kit—an automatic microarray assay based on reverse hybridization—for the detection of ARGs directly from positive blood culture (PBC) samples. A total of 111 Gram-negative bacterial isolates (92 Enterobacterales, 14 Acinetobacter baumannii, and 6 Pseudomonas spp.), previously characterized by whole-genome sequencing (WGS), were each used to generate a PBC, which was then analyzed with the HS12a/MDR-FC assay. Results: We demonstrated perfect agreement for the detection of macrolide resistance genes across all bacterial species and high agreement for genes conferring resistance to sulfonamides and β-lactams. In contrast, aminoglycoside resistance genes showed only moderate agreement, with minor discrepancies observed in Klebsiella pneumoniae and Escherichia coli, largely attributable to specific SNP variations. Conclusions: The HS12a/MDR-FC assay includes 51 ARGs, though not all were represented in our isolate set, and some false negatives were observed. Despite these limitations, its broad coverage and rapid turnaround remain advantageous compared to other rapid assays with fewer targets. Future refinements should aim at broader gene coverage, inclusion of key mutations, and detection of emerging variants, making this approach a promising tool for rapid AMR surveillance and antimicrobial stewardship. Full article

Rapid and reliable on-site identification of body fluids is essential in forensic and field diagnostic applications. Commercial kits provide only single results and often suffer from cross-reactivity, while conventional microarrays offer multiplex capability but lack sufficient fluorescence intensity for field-deployable systems. In this study, we present a highly sensitive nanorods on micro post array (NMPA) substrate and a smartphone-based portable detection system. The NMPA substrate integrates metal nanorods with UV-imprinted micro post structures to produce metal-enhanced fluorescence and improved signal localization. When evaluated using a microarray scanner, the substrate achieved high sensitivity, detecting semen diluted up to 1/100,000. The portable smartphone system further demonstrated simultaneous detection of three semen biomarkers (PSA, ACPP, and Semenogelin-1) at a 1/1000 dilution, matching the detection limit of commercial kits. Specificity tests using blood, saliva, urine, vaginal fluid, and environmental contaminants showed no false-positive responses. These results highlight the potential of the NMPA system as a portable diagnostic technology capable of rapid (<15 min), multiplex, and highly sensitive detection in field environments. Future work will focus on quantitative calibration, substrate stability assessment, and expansion toward multi biomarker panels for broader forensic and clinical applications. Full article

African swine fever is a highly contagious and deadly disease of both domestic and wild pigs. In developing countries such as Ghana, the diagnosis and control of ASF are very challenging. In this paper, we discussed factors that account for ASF endemicity in many developing nations, with special focus on Ghana. We identified possible ASF spread via pig value chain through the transportation of live pigs across regions in Ghana. Major factors contributing to ASF spread in Ghana include lack of farmer compensation during ASF outbreaks, free range system of pig farming, porous country borders, lack of rapid on-site diagnostic test kits, unsafe sample collection and transportation to diagnostic laboratories, long diagnostic test turnaround, and improper carcass disposal. We also discuss available diagnostic options for ASF and their limitations. We propose a more holistic approach to ASF control in Ghana. These measures include applying a muti-sectoral approach, rehabilitation of inactive regional laboratories and expansion of services to six newly established regions, promoting point of care testing and developing and implementing farmer compensation plan during outbreaks. These proposed ASF control measures will provide field veterinarians with effective means to make informed decisions while awaiting laboratory confirmation of outbreaks. Full article

Acute gastroenteritis (AGE) is a major cause of illness and death in children under five, especially in low- and middle-income countries, and rotavirus A (RVA) and norovirus are the leading viral agents. The present study aimed to describe the development of a commercial multiplex TaqMan-based RT-qPCR assay to detect those viruses to enhance surveillance and public health responses in Brazil. The assay validation involved optimizing primers and probes for multiplex RT-qPCR, assessing analytical sensitivity, and confirming specificity. A multicenter pilot study across Brazil’s AGE surveillance network assessed the assay’s performance. The IBMP NAT assay demonstrated high specificity and sensitivity for detecting RVA and norovirus GI and GII. No cross-reactivity was observed. LoD95 values were low: 18.6 (GI), 71.2 (GII), and 12.3 (RVA) copies/reaction. In 379 clinical samples, diagnostic sensitivity and specificity exceeded 96% for all targets. The assay showed strong reproducibility across operators and instruments. Stability tests confirmed consistent performance under freeze–thaw, transport, and storage conditions. Compared to in-house RT-qPCR, the IBMP NAT test yielded lower Ct values, indicating improved detection of low viral loads. The IBMP NAT Kit significantly advances molecular diagnostics, enabling rapid, sensitive, and reliable detection of RVA and norovirus in fecal specimens. It strengthens public health surveillance and supports timely responses to AGE outbreaks, helping reduce disease burden in vulnerable populations. Full article

Accurate pathogen detection in bovine semen is crucial for animal health surveillance and international trade. Semen presents unique challenges due to the presence of PCR inhibitors from seminal plasma and extender components, reducing nucleic acid extraction efficiency and sensitivity. The two National Animal Health Laboratory Network-approved extraction platforms (MagMAX CORE and IndiMag Pathogen Kits) were evaluated using 88 negative extended semen samples at 200 µL input volume, reduced input volumes, and pretreatment strategies with two influenza A virus (IAV) PCR assays, containing different exogenous internal controls (ICs) to assess PCR inhibition. The ICs yielded overall passing rates from 31.8% to 100.0% and varied greatly based on the extender formulation and extraction protocol. Validation continued with naturally infected semen containing Mycoplasma bovis, bovine viral diarrhea virus, bovine herpesvirus-1, and the limit of detection using Mycoplasma bovis. The IndiMag Pathogen 100-na was then selected for evaluation of diagnostic sensitivity and specificity, reproducibility, and detection limits with IAV-spiked samples, using the two IAV PCR assays and their ICs. Selected archived semen samples used in this study were screened and were negative for IAV by both PCR assays. These findings underscore the importance of tailored extraction methods in overcoming semen-associated inhibition and facilitating reliable pathogen surveillance in bovine germplasm. Full article