Search Results (343)

The global spread of Mpox virus (MPXV) underscores the urgent need for rapid, field-deployable diagnostic tools, especially in low-resource settings. We evaluated a loop-mediated isothermal amplification (LAMP) assay, termed LAMPOX, developed by Coris BioConcept. The assay was tested in three formats—two liquid versions and a dried, ready-to-use version—targeting only the ORF F3L (Liquid V1) or both the ORF F3L and N4R (Liquid V2 and dried) genomic regions. Analytical sensitivity and specificity were assessed using 60 clinical samples from confirmed MPXV-positive patients. Sensitivity on clinical samples was 81.7% for Liquid V1 and 88.3% for Liquid V2. The dried LAMPOX assay demonstrated a sensitivity of 88.3% and a specificity of 100% in a panel of 112 negative controls, with most positive samples detected in under 7 min. Additionally, a simplified sample lysis protocol was developed to facilitate point-of-care use. While this method showed slightly reduced sensitivity compared to standard DNA extraction, it proved effective for samples with higher viral loads. The dried format offers key advantages, including ambient-temperature stability and minimal equipment needs, making it suitable for point-of-care testing. These findings support LAMPOX as a promising tool for rapid MPXV detection during outbreaks, especially in resource-limited settings where traditional PCR is impractical. Full article

Porcine Rotavirus (PoRV), a predominant causative agent of neonatal diarrhea in piglets, shares substantial genetic homology with human rotavirus and represents a considerable threat to both public health and the global swine industry in the absence of specific antiviral interventions. The VP6 protein, an internal capsid component, is characterized by exceptional sequence conservation and robust immunogenicity, rendering it an ideal candidate for viral genotyping and vaccine development. In the present study, the recombinant plasmid pET28a(+)-VP6 was engineered to facilitate the high-yield expression and purification of the VP6 antigen. BALB/c mice were immunized to generate monoclonal antibodies (mAbs) through hybridoma technology, and the antigenic specificity of the resulting mAbs was stringently validated. Subsequently, a panel of truncated protein constructs was designed to precisely map linear B-cell epitopes, followed by comparative conservation analysis across diverse PoRV strains. Functional validation demonstrated that all three mAbs exhibited high-affinity binding to VP6, with a peak detection titer of 1:3,000,000 and exclusive specificity toward PoRVA. These antibodies effectively recognized representative genotypes such as G3 and X1, while exhibiting no cross-reactivity with unrelated viral pathogens; however, their reactivity against other PoRV serogroups (e.g., types B and C) remains to be further elucidated. Epitope mapping identified two novel linear B-cell epitopes, ¹²⁸YIKNWNLQNR¹³⁷ and ¹³⁸RQRTGFVFHK¹⁴⁷, both displaying strong sequence conservation among circulating PoRV strains. Collectively, these findings provide a rigorous experimental framework for the functional dissection of VP6 and reinforce its potential as a valuable diagnostic and immunoprophylactic target in PoRV control strategies. Full article

Infectious diseases poses a growing public health challenge. The COVID-19 pandemic has further emphasized the urgent need for rapid, accessible diagnostics. This study presents the development of an integrated, flexible point-of-care (POC) diagnostic system for the rapid detection of Corynebacterium diphtheriae, the pathogen responsible for diphtheria. The system comprises a microfluidic polymerase chain reaction (micro-PCR) device and an electrochemical DNA biosensor, both fabricated on flexible substrates. The micro-PCR platform offers rapid DNA amplification overcoming the time limitations of conventional thermocyclers. The biosensor utilizes specific molecular recognition and an electrochemical transducer to detect the amplified DNA fragment, providing a clear and direct indication of the pathogen’s presence. The combined system demonstrates the effective amplification and detection of a gene fragment from a toxic strain of C. diphtheriae, chosen due to its increasing incidence. The design leverages lab-on-a-chip (LOC) and microfluidic technologies to minimize reagent use, reduce cost, and support portability. Key challenges in microsystem design—such as flow control, material selection, and reagent compatibility—were addressed through optimized fabrication techniques and system integration. This work highlights the feasibility of using flexible, integrated microfluidic and biosensor platforms for the rapid, on-site detection of infectious agents. The modular and scalable nature of the system suggests potential for adaptation to a wide range of pathogens, supporting broader applications in global health diagnostics. The approach provides a promising foundation for next-generation POC diagnostic tools. Full article

Point-of-care (POC) diagnostic technologies have become essential for the real-time monitoring and management of chronic wounds, where maintaining a moist environment and controlling pH levels are critical for effective healing. In this study, a flexible pH sensor based on a graphene/molybdenum disulfide (graphene/MoS₂) composite interdigitated electrode (IDE) structure was fabricated using pulsed laser ablation. The pH sensor, with an active area of 30 mm × 30 mm, exhibited good adhesion to the polyethylene terephthalate (PET) substrate and maintained structural integrity under repeated bending cycles. Precise ablation was achieved under optimized conditions of 4.35 J/cm² laser fluence, a repetition rate of 300 kHz, and a scanning speed of 500 mm/s, enabling the formation of defect-free IDE arrays without substrate damage. The influence of laser processing parameters on the surface morphology, electrical conductivity, and wettability of the composite thin films was systematically characterized. The fabricated pH sensor exhibited high sensitivity (~4.7% change in current per pH unit) across the pH 2–10 range, rapid response within ~5.2 s, and excellent mechanical stability under 100 bending cycles with negligible performance degradation. Moreover, the sensor retained > 95% of its stable sensitivity after 7 days of ambient storage. Furthermore, the pH response behavior was evaluated for electrode structures with different pitches, demonstrating that structural design parameters critically impact sensing performance. These results offer valuable insights into the scalable fabrication of flexible, wearable pH sensors, with promising applications in wound monitoring and personalized healthcare systems. Full article

Porcine rotavirus (PoRV), a primary etiological agent of viral diarrhea in piglets, frequently co-infects with other enteric pathogens, exacerbating disease severity and causing substantial economic losses. Its genetic recombination capability enables cross-species transmission potential, posing public health risks. Globally, twelve G genotypes and thirteen P genotypes have been identified, with G9, G5, G3, and G4 emerging as predominant circulating strains. The limited cross-protective immunity between genotypes compromises vaccine efficacy, necessitating genotype surveillance to guide vaccine development. While conventional molecular assays demonstrate sensitivity, they lack rapid genotyping capacity and face technical limitations. To address this, we developed a novel diagnostic platform integrating reverse transcription recombinase-aided amplification (RT-RAA) with CRISPR–Cas12a. This system employs universal primers for the simultaneous amplification of G4/G5/G9 genotypes in a single reaction, coupled with sequence-specific CRISPR recognition, achieving genotyping within 50 min at 37 °C with 10⁰ copies/μL sensitivity. Clinical validation showed a high concordance with reverse transcription quantitative polymerase chain reaction (RT-qPCR). This advancement provides an efficient tool for rapid viral genotyping, vaccine compatibility evaluation, and optimized epidemic control strategies. Full article

The Internet of Medical Things (IoMT) is revolutionizing healthcare by integrating smart diagnostic devices with cloud computing and real-time data analytics. The emergence of infectious diseases, including COVID-19, underscores the need for rapid and decentralized diagnostics to facilitate early intervention. Traditional centralized laboratory testing introduces delays, limiting timely medical responses. While point-of-care molecular diagnostic (POC-MD) systems offer an alternative, challenges remain in cost, accessibility, and network inefficiencies. This study proposes an IoMT-based architecture for fully automated POC-MD devices, leveraging WebSockets for optimized communication, enhancing microfluidic cartridge efficiency, and integrating a hardware-based emulator for real-time validation. The system incorporates DNA extraction and real-time polymerase chain reaction functionalities into modular, networked components, improving flexibility and scalability. Although the system itself has not yet undergone clinical validation, it builds upon the core cartridge and detection architecture of a previously validated cartridge-based platform for Chlamydia trachomatis and Neisseria gonorrhoeae (CT/NG). These pathogens were selected due to their global prevalence, high asymptomatic transmission rates, and clinical importance in reproductive health. In a previous clinical study involving 510 patient specimens, the system demonstrated high concordance with a commercial assay with limits of detection below 10 copies/μL, supporting the feasibility of this architecture for point-of-care molecular diagnostics. By addressing existing limitations, this system establishes a new standard for next-generation diagnostics, ensuring rapid, reliable, and accessible disease detection. Full article

The COVID-19 pandemic and recent viral outbreaks have highlighted the need for viral diagnostics that balance accuracy with accessibility. While traditional laboratory methods remain essential, point-of-care solutions are critical for decentralized testing at the population level. However, a gap persists between academic proof-of-concept studies and clinically viable tools, with novel technologies remaining inaccessible to clinics due to cost, complexity, training, and logistical constraints. Recent advances in surface functionalization, assay simplification, multiplexing, and performance in complex media have improved the feasibility of both optical and non-optical sensing techniques. These innovations, coupled with scalable manufacturing methods such as 3D printing and streamlined hardware production, pave the way for practical deployment in real-world settings. Additionally, software-assisted data interpretation, through simplified readouts, smartphone integration, and machine learning, enables the broader use of diagnostics once limited to experts. This review explores improvements in viral diagnostic approaches, including colorimetric, optical, and electrochemical assays, showcasing their potential for democratization efforts targeting the clinic. We also examine trends such as open-source hardware, modular assay design, and standardized reporting, which collectively reduce barriers to clinical adoption and the public dissemination of information. By analyzing these interdisciplinary advances, we demonstrate how emerging technologies can mature into accessible, low-cost diagnostic tools for widespread testing. Full article

Accurate, rapid and inexpensive diagnosis of poultry respiratory pathogens remains a challenge, especially in many developing countries. Meanwhile, poultry respiratory pathogens are a major threat to poultry production worldwide, accounting for billions of dollars in economic loss to the sector. Early and accurate diagnosis of these diseases is critical for economic poultry production. Molecular diagnostic methods, including PCR-based techniques, have been developed and used to fill this gap, but unfortunately, these techniques require skilled technicians, relatively costly equipment and reagents and can only be performed in a laboratory setting. This warrants the development of other diagnostic tools, which can be used in the field even by unskilled personnel. In this review, we discussed the genesis, challenges, advances and prospects of loop-mediated isothermal amplification (LAMP) for the detection of poultry respiratory pathogens at the flock side, especially in resource-constrained countries. We highlighted the application of LAMP in routine poultry disease surveillance and early outbreak detection, underscoring its value as a transformative diagnostic tool in poultry production. The development and use of a point-of-care (POC) LAMP assay that can be used to screen for these poultry respiratory pathogens simultaneously enhance disease surveillance and diagnosis. Full article

Escherichia coli (E. coli) remains a major concern in poultry production due to its ability to incite foodborne illness and public health crisis, zoonotic potential, and the increasing prevalence of antibiotic-resistant strains. The contamination of poultry products with pathogenic E. coli, including avian pathogenic E. coli (APEC) and Shiga toxin-producing E. coli (STEC), presents risks at multiple stages of the poultry production cycle. The stages affected by E. coli range from, but are not limited to, the hatcheries to grow-out operations, slaughterhouses, and retail markets. While traditional detection methods such as culture-based assays and polymerase chain reaction (PCR) are well-established for E. coli detection in the food supply chain, their time, cost, and high infrastructure demands limit their suitability for rapid and field-based surveillance—hindering the ability for effective cessation and handling of outbreaks. Biosensors have emerged as powerful diagnostic tools that offer rapid, sensitive, and cost-effective alternatives for E. coli detection across various stages of poultry development and processing where detection is needed. This review examines current biosensor technologies designed to detect bacterial biomarkers, toxins, antibiotic resistance genes, and host immune response indicators for E. coli. Emphasis is placed on field-deployable and point-of-care (POC) platforms capable of integrating into poultry production environments. In addition to enhancing early pathogen detection, biosensors support antimicrobial resistance monitoring, facilitate integration into Hazard Analysis Critical Control Points (HACCP) systems, and align with the One Health framework by improving both animal and public health outcomes. Their strategic implementation in slaughterhouse quality control and marketplace testing can significantly reduce contamination risk and strengthen traceability in the poultry value chain. As biosensor technology continues to evolve, its application in E. coli surveillance is poised to play a transformative role in sustainable poultry production and global food safety. Full article

Sepsis remains a global health emergency, demanding timely and accurate diagnostics to reduce morbidity and mortality. This review critically assesses the recent progress (2020–2025) in the development of electrochemical aptamer-based biosensors for sepsis detection. These biosensors combine aptamers’ high specificity and modifiability with the sensitivity and miniaturization potential of electrochemical platforms. The analysis highlights notable advances in detecting key sepsis biomarkers, such as C-reactive protein (CRP), procalcitonin (PCT), interleukins (e.g., interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α)), lipopolysaccharides (LPSs), and microRNAs using diverse sensor configurations, including a field-effect transistor (FET), impedance spectroscopy, voltammetry, and hybrid nanomaterial-based systems. A comparative evaluation reveals promising analytical performance in terms of the limit of detection (LOD), rapid response time, and point-of-care (POC) potential. However, critical limitations remain, including variability in validation protocols, limited testing in real clinical matrices, and challenges in achieving multiplexed detection. This review underscores translational barriers and recommends future directions focused on clinical validation, integration with portable diagnostics, and interdisciplinary collaboration. By consolidating current developments and gaps, this work provides a foundation for guiding next-generation biosensor innovations aimed at effective sepsis diagnosis and monitoring. Full article

Background/Objectives: Syphilis remains a significant public health challenge worldwide. Accurate and efficient diagnostic tools are essential to controlling the spread of the disease. Current diagnostic approaches primarily rely on serologic treponemal tests (TTs) and nontreponemal tests (NTTs). The aim of this study was to evaluate the diagnostic properties of various serological methods for syphilis diagnosis. Methods: Samples were collected from participants of the Health, Information, and Sexually Transmitted Infection Monitoring (SIM study) between March 2020 and May 2023, using convenience sampling at a mobile health unit in Porto Alegre, Brazil. A total of 250 individuals were tested using the point-of-care (POC) lateral flow treponemal test, Venereal Disease Research Laboratory (VDRL) test, Rapid Plasma Reagin (RPR) test, Enzyme-Linked Immunosorbent Assay (ELISA), and Treponema pallidum hemagglutination assay (TPHA). Of these, 125 participants tested positive for syphilis in the POC screening. Diagnostic properties such as sensitivity, specificity, and predictive values were assessed for the POC test, ELISA, and VDRL test. The TPHA was used as the reference standard for the TT, and the RPR test as the reference standard for the NTT. Results: Among individuals with positive POC test results, 97.6% (122/125) were also positive by the ELISA, and 85.6% (107/125) were positive by the TPHA. Additionally, 48.0% (60/125) and 42.4% (53/125) tested positive by the VDRL and RPR tests, respectively. Using the TPHA as a reference, TT tests showed sensitivities of 97–98% and specificities of 93–95% for detecting anti-Treponema pallidum antibodies using the ELISA and POC test, respectively. For the NTT, the VDRL test demonstrated a sensitivity of 98% and a specificity of 95% compared to the RPR test. The kappa coefficients were 0.85 for the POC test vs. the TPHA, 0.81 for the ELISA vs. the TPHA, and 0.89 for the VDRL vs. the RPR tests, indicating substantial agreement. Conclusions: This study highlights a good diagnostic performance and high agreement levels among the evaluated serological tests for syphilis, reinforcing their utility in clinical and public health settings, as well as epidemiological studies. Full article

Background/Objective: Early pregnancy loss is often caused by chromosomal abnormalities, necessitating accurate diagnostic tools. While product of conception (POC) chromosomal testing is commonly used, it can be limited by culture failure or an inability to obtain fetal tissue due to spontaneous expulsion. Cell-free fetal DNA (cff DNA) analysis provides a non-invasive alternative; however, its effectiveness in early pregnancy loss, particularly in cases where fetal components are still minimal, has not been fully established. The objective of this study was to evaluate the accuracy of pre- and postoperative cff DNA analysis for detecting fetal aneuploidy by comparing the results to those of POC chromosomal testing. Methods: In this single-center prospective cohort study, 50 women undergoing manual vacuum aspiration for pregnancy loss before 12 weeks of gestation were enrolled (February 2022–December 2024). Cff DNA analysis was performed on maternal blood samples collected pre- and postoperatively. The primary outcome was concordance between the cff DNA and POC results. Sensitivity, specificity, and factors affecting concordance were also assessed. Results: Eight participants were excluded due to unsuccessful POC culture (n = 3), suspected maternal tissue contamination in the POC sample (n = 1), mosaicism (n = 3), or triploidy (n = 1), resulting in 42 evaluable cases. Preoperative cff DNA analysis showed 88.1% concordance with POC (sensitivity 86.4% and specificity 90.0%). Postoperative analysis showed 78.6% concordance (sensitivity 72.7% and specificity 85.0%). Conclusions: The Cff DNA analysis of preoperative and postoperative maternal blood samples showed generally good concordance with conventional POC chromosomal testing in detecting fetal aneuploidy in early pregnancy loss. Full article

Background/Objectives: Gram staining is an essential diagnostic technique used for the rapid identification of bacterial and fungal infections, playing a pivotal role in clinical decision-making, especially in point-of-care (POC) settings. Manual staining, while effective, is labor-intensive and prone to variability, relying heavily on the skill of laboratory personnel. Current automated Gram-staining systems are primarily designed for high-throughput laboratory environments, limiting their feasibility in decentralized healthcare settings such as emergency departments and rural clinics. This study aims to introduce and evaluate the Point-of-Care Gram Stainer (PoCGS^®), a compact, automated device engineered for single-slide processing, addressing challenges related to portability, standardization, and efficiency in POC applications. Methods: The PoCGS^® device was developed to emulate expert manual staining techniques through features such as methanol fixation and programmable reagent application. A comparative evaluation was performed using 40 urine samples, which included both clinical and artificial specimens. These samples were processed using PoCGS^®, manual staining by skilled experts, and manual staining by unskilled personnel. The outcomes were assessed based on microbial identification concordance, the staining uniformity, presence of artifacts, and agreement with the culture results. Statistical analyses, including agreement rates and quality scoring, were conducted to compare the performance of PoCGS^® against manual staining methods. Results: PoCGS^® achieved a 100% concordance rate with expert manual staining in terms of microbial identification, confirming its diagnostic accuracy. However, staining quality parameters such as the uniformity and presence of artifacts showed statistically significant differences when compared to skilled and unskilled personnel. Despite these limitations, PoCGS^® demonstrated a comparable performance regarding artifact reduction and agreement with the culture results, indicating its potential utility in POC environments. Challenges such as fixed processing times and limited adaptability to varying specimen characteristics were identified as areas for further improvement. Conclusions: The study findings suggest that PoCGS^® is a reliable and valuable tool for microbial identification in POC settings, with a performance comparable to skilled manual staining. Its compact design, automation, and ease of use make it particularly beneficial for resource-limited environments. Although improvements in staining uniformity and background clarity are required, PoCGS^® has the potential to standardize Gram staining protocols and improve diagnostic turnaround times. Future developments will focus on optimizing staining parameters and expanding its application to other clinical sample types, ensuring robustness and broader usability in diverse healthcare settings. Full article

Porcine epidemic diarrhea virus (PEDV), a primary pathogen causing diarrhea in pigs, particularly in piglets, has a mortality rate of up to 100%. Field PEDV strains circulating in pig production can be phylogenetically divided into two genotypes, GI and GII. Differential diagnosis of clinical strains with different genotypes is helpful for understanding disease epidemiology, vaccine selection, and prevention and control measures. The loop-mediated isothermal amplification method (LAMP), a novel nucleic acid amplification technique, has been utilized to detect a variety of pathogens in practice. In this study, we developed a distinguished RT-LAMP method to identify genotypes GI and GII strains of PEDV. Two pairs of primers, PEDV-LM and PEDV-LS, were designed based on the membrane and spike genes of PEDV, respectively. PEDV-LM primers exhibited specificity for all PEDV strains, while PEDV-LS primers specifically targeted the PEDV genotype GI. The diagnostic sensitivity of both primers was 1 × 10² copies/reaction, which is 100 times more sensitive than RT-PCR. The RT-LAMP reaction process was completed at 65 °C for 40 min just in a water bath or metal bath. A cross-reactivity assay confirmed that this method is specific for PEDV GI and GII, with no cross-amplification observed with other swine-origin viruses such as PDCoV, PoRV, PRV, and PRRSV. Therefore, this refined LAMP technique offers a rapid, sensitive, and reliable method with which to detect and differentiate between PEDV GI and GII, making it a superior tool for the large-scale clinical surveillance of PEDV infections. Full article

The development of cost-effective, rapid-response, and user-friendly biosensing platforms has become paramount importance for achieving precise biomarker quantification in early disease detection. Implementing timely diagnostic interventions through accurate biomarker analysis not only significantly improves treatment outcomes but also enables effective disease management strategies, ultimately leading to substantial reductions in patient mortality rates. These clinical imperatives have consequently driven the innovation of portable point-of-care (POC) diagnostic systems. Electrochemical biosensors are attractive in the early diagnosis of diseases due to their low cost, simple operation, and high sensitivity. This review examines prevalent material innovations in electrode functionalization for electrochemical biosensing platforms, with specific emphasis on their translational applications in early-stage disease detection. The analysis included three important early diagnostic biomarker types: proteins, nucleic acids, and small molecule metabolites. Furthermore, the work proposes novel research trajectories for next-generation biosensor development, advocating the synergistic integration of artificial intelligence-driven analytics, Internet of Medical Things (IoMT)-enabled diagnostic networks, and advanced micro/nanofabrication techniques. Full article