Mycobacterial lung diseases, including tuberculosis and nontuberculous mycobacterial infections, pose a significant global health threat amplified by rising antimicrobial resistance. Traditional drug development approaches using two-dimensional cell cultures and animal models frequently fail to accurately predict drug efficacy and lung penetration, creating critical bottlenecks in therapeutic discovery. To address this translational gap, researchers are adopting New Approach Methodologies (NAMs)—advanced technologies including organoids, air-liquid interface systems, and organ-on-chip platforms that more accurately replicate human lung physiology and host-pathogen interactions. These microphysiological systems enable evaluation of drug distribution, bacterial persistence monitoring, and identification of predictive biomarkers for antibiotic effectiveness. This research initiative establishes an interdisciplinary platform integrating microfluidics, biomaterials, pharmacology, and microbiology to accelerate antibiotic development and optimize treatment strategies. The effort solicits original research and reviews focusing on evaluating novel antibiotics using lung organoids, modeling resistance mechanisms in Mycobacterium tuberculosis and nontuberculous species, identifying efficacy biomarkers through multi-omics approaches, assessing pharmacokinetics in engineered tissue models, and validating NAM-based infection models for high-throughput screening. This comprehensive approach promises to enhance drug discovery efficiency and facilitate development of host-directed therapies against mycobacterial pathogens.

Pseudomonas aeruginosa is a multidrug resistant gram-negative bacterium that poses a significant global health threat, particularly for immunocompromised patients. The organism employs diverse resistance mechanisms including efflux pump overexpression, porin modification, enzymatic inactivation, and target site modifications, alongside phenotypic adaptations such as biofilm and persister cell formation. These complex strategies have driven the emergence of multidrug resistant, extensively drug resistant, and pan drug resistant strains, complicating treatment options. While conventional culture-based diagnostics remain the gold standard, they suffer from delays and limitations in detecting heteroresistance and biofilm-associated tolerance. Recent advances in omics-based approaches—including genomics, epigenomics, transcriptomics, proteomics, lipidomics, metabolomics, and phenomics—offer rapid and precise alternatives for identifying resistant P. aeruginosa. Complementary innovations such as microfluidic lab-on-chip systems and machine learning-driven artificial intelligence further enhance diagnostic capabilities. Integrating multi-omics data with advanced platforms represents a revolutionary strategy for comprehensive, rapid resistance profiling in precision diagnostics. However, standardized protocols, clinical validation, and cost-effective implementation are urgently required to translate this potential into clinical practice and reduce the global antimicrobial resistance burden.

Antimicrobial resistance poses a critical global health crisis, causing over 1.2 million deaths annually with projections reaching 10 million yearly by 2050. Bacteriophage therapy has emerged as a promising alternative to combat multidrug-resistant bacteria, including ESKAPE pathogens, and is advancing through controlled clinical trials supported by the WHO and TATFAR. However, clinical translation remains constrained by the lack of rapid, standardized methods for therapeutic phage selection. Researchers have developed digital phage susceptibility testing (dPhaST), an automated droplet digital PCR workflow that quantifies phage-induced DNA release as a marker of bacterial lysis by targeting conserved 16S rRNA regions. Testing 122 phage-host combinations across six bacterial species, dPhaST achieved 95.9% concordance with traditional spot tests while completing analysis within three hours and resolving weak lytic activities that conventional methods miss. The approach proves robust against phage-encoded nucleases and cross-contamination, capturing defense mechanisms involving CRISPR-Cas and Sir2-HerA systems. This automated quantification method enables rapid, mechanistically informative profiling of phage efficacy across diverse pathogens, significantly advancing phage therapy's clinical potential.

# Summary This review examines optical biosensors as diagnostic tools for viral infectious diseases, providing a comprehensive analysis of their current clinical applications and future potential. Optical biosensors leverage light-based detection mechanisms to identify viral pathogens with high sensitivity and specificity, offering significant advantages over conventional diagnostic methods. The review covers various optical technologies including surface plasmon resonance, fluorescence-based assays, and photonic devices that enable rapid, label-free or minimally invasive virus detection. The analysis encompasses both established viral infections and emerging pathogens, highlighting the role of optical biosensors in point-of-care diagnostics and laboratory settings. Key considerations include assay sensitivity, specificity, cost-effectiveness, and integration with portable platforms. The review emphasizes how these technologies address critical diagnostic challenges such as reducing detection time and improving accessibility in resource-limited settings. Future perspectives focus on nanotechnology integration, multiplexing capabilities, and real-time monitoring applications. Optical biosensors represent a transformative approach to infectious disease diagnosis, with significant implications for pandemic preparedness, early detection strategies, and personalized medicine in virology.

Dr. Zubair Khalid, a veterinarian and virologist, specializes in advancing diagnostic and therapeutic innovations through molecular virology expertise. His research focuses on leveraging CRISPR-Cas12a-based nucleic acid detection systems for rapid pathogen diagnosis, addressing critical gaps in conventional diagnostic methods. This approach represents a significant technological advancement, as CRISPR-Cas12a offers specificity and speed superior to traditional techniques, enabling faster identification of viral and infectious agents in clinical and veterinary settings. Dr. Khalid's work integrates multiple disciplines—conventional virology, molecular biology, and computational approaches—to enhance diagnostic accuracy and accessibility. His expertise in vaccine development and multi-omics analysis further strengthens the translational potential of these innovations. By combining cutting-edge molecular techniques with veterinary science, his research demonstrates substantial promise for improving animal health outcomes and supporting rapid response capabilities during disease outbreaks. This interdisciplinary approach underscores the growing importance of molecular diagnostics in contemporary infectious disease management.

The rapid advancement in personalized healthcare has driven the development of wearable biomedical devices for real-time biomarker monitoring and diagnosis. Traditional invasive blood-based diagnostics are painful and limited to sporadic health snapshots. To address these limitations, microneedle-based sensing platforms have emerged, utilizing interstitial fluid (ISF) as an alternative biofluid for continuous health monitoring in a minimally invasive and painless manner. This review aims to provide a comprehensive overview of microneedle sensor technology, covering microneedle design, fabrication methods, and sensing strategy. Additionally, it explores the integration of monitoring electronics for continuous on-body monitoring. Representative applications of microneedle sensing platforms for both monitoring and therapeutic purposes are introduced, highlighting their potential to revolutionize personalized healthcare. Finally, the review discusses the remaining challenges and future prospects of microneedle technology.

Reliable and continuous access to interstitial fluid (ISF) remains a major challenge for wearable and tissue-embedded biosensing systems. Conventional microneedle-based methods, while widely adopted, often exhibit variable sampling efficiency across different skin types and are prone to performance fluctuations during motion. Herein, we introduce a tissue-embedded thread-based open capillary microfluidics-based sampling that enables minimally invasive, pump-free, and reliable continuous monitoring of small molecules from ISF. The system employs open capillary-driven microfluidic channels in textile threads, which facilitate the partial separation of small molecules from complex biological matrices, mimicking a microdialysis process. The continuous sampling is achieved through evaporation-driven capillary pressure, eliminating the need for an external pump. The experimental results confirm the numerical simulation of diffusion-controlled and capillary-driven transport of analytes in ISF.

Light-enhanced molecular diagnostic technologies are advancing infectious disease detection by overcoming limitations of conventional methods, which often suffer from weak signal strength, slow processing, and difficulty detecting low-abundance targets in complex samples. These innovative approaches leverage plasmonics, photonics, and nano-optical effects to enable enhanced pathogen identification and antimicrobial resistance profiling. Recent studies demonstrate significant progress in fluorescence, chemiluminescence, surface-enhanced Raman scattering (SERS), and photoacoustic applications for sensitive pathogen detection. However, challenges remain in reproducibility, device integration simplification, and real-world clinical validation, particularly for translating laboratory-based optical platforms into point-of-care systems suitable for resource-limited healthcare settings. A comprehensive research initiative aims to strengthen the theoretical and technical foundations of optical signal amplification while fostering clinical implementation of light-enhanced diagnostics. Priority areas include investigating light-matter interactions at nano- and microscales to amplify biological recognition, integrating optical sensing into compact diagnostic platforms, and improving pathogen detection, antimicrobial resistance monitoring, and outbreak response capabilities. The initiative welcomes research on light-induced signal amplification, fluorescence-based platforms, plasmonic materials, photothermal and photoacoustic detection strategies, and microfluidic integration to advance infection diagnostics.

Isothermal nucleic acid amplification technologies (INAATs) represent a transformative alternative to polymerase chain reaction (PCR) for infectious disease diagnostics, addressing critical limitations in resource-limited and decentralized settings. Unlike PCR, which requires thermal cycling equipment and specialized laboratory infrastructure, INAATs including LAMP, RPA, RAA, MIRA, HDA, RCA, SDA, and NASBA enable nucleic acid amplification at constant temperature while maintaining high sensitivity and specificity. These methods support detection of bacterial, viral, fungal, and parasitic pathogens without sophisticated instrumentation. Recent advances in enzyme engineering, assay design, CRISPR-based detection, biosensors, microfluidics, and portable devices have significantly enhanced performance capabilities. This research initiative compiles the latest innovations across methodological developments and clinical applications, including point-of-care diagnostics for outbreak response, clinical diagnosis, and surveillance. The compilation welcomes contributions spanning enzyme optimization, novel amplification chemistries, integration with next-generation detection technologies, and veterinary, environmental, and food safety applications, enabling the transition from conventional laboratory testing to accessible diagnostic platforms where infectious disease burden is greatest.

Researchers have developed a pocket-sized point-of-care testing platform that eliminates the need for electricity, complex laboratory equipment, and skilled technicians. The fully integrated device uses a manual suction pump with a compression spring to operate fluidic functions without external power, achieving high reproducibility with coefficients of variation below 4.5 percent. The platform incorporates a serpentine microchannel design that leverages Dean flow-induced inertial mixing to accelerate molecular aggregation, enabling a wash-free plasmonic assay. Validated with two critical biomarkers—thrombin for cardiovascular and renal diagnostics, and SARS-CoV-2 spike protein for viral screening—the system delivers quantitative results within five minutes with sub-nanomolar detection limits of 0.5 nanomolar for thrombin and 0.4 nanomolar for spike protein. Wireless smartphone connectivity enables immediate data visualization. This innovation addresses critical gaps in decentralized diagnostics, particularly for resource-limited settings and developing countries where conventional laboratory infrastructure is unavailable, while responding to demonstrated pandemic-era demands for rapid, portable diagnostic solutions.

Researchers from the Reches group at The Hebrew University of Jerusalem have published significant findings on peptide coacervates in ACS Applied Materials and Interfaces, advancing understanding of a fundamental biological process with disease implications. Coacervates are condensates formed through liquid-liquid phase separation (LLPS), a process where solutes self-concentrate into microdroplets that function as membraneless organelles within cells. This phenomenon underlies multiple biological processes and is implicated in neurodegenerative diseases including Alzheimer's, Parkinson's, and Huntington's disease. The research characterizes coacervate behavior and demonstrates how their tailored design could enhance control, encapsulation efficiency, and antioxidant properties for therapeutic applications. By modulating peptide coacervates, researchers can potentially develop improved drug delivery systems and disease treatments. The work builds on foundational research examining how genetic mutations affecting protein solubility or regulatory mechanisms, along with altered physiological conditions, can trigger abnormal LLPS leading to pathological disease states. This research bridges chemistry and therapeutics, offering promising avenues for targeted disease intervention.

Sepsis treatment relies critically on rapid administration of appropriate antibiotics, yet current clinical protocols for selecting patient-specific therapy remain inefficiently slow due to prolonged blood culture procedures. Researchers have developed an ultra-rapid antimicrobial susceptibility testing (AST) method that eliminates the traditional blood culture requirement, potentially reducing reporting turnaround time by 40 to 60 hours compared to standard hospital workflows. The innovation introduces a synthetic beta-2-glycoprotein I peptide that selectively recovers diverse microbial pathogens directly from whole blood, enabling immediate species identification and phenotypic drug evaluation using a low-inoculum AST chip. A clinical evaluation involving 190 hospitalized patients with suspected infections demonstrated 100 percent accuracy in species identification. Among eight positive cases, six clinical isolates showed 94.90 percent categorical agreement with conventional AST, achieving an average theoretical turnaround time of just 13 hours from initial blood processing. This blood culture-free approach addresses a major clinical bottleneck in sepsis management, offering significant potential to accelerate treatment decisions and improve patient outcomes by enabling faster antimicrobial selection.

Candidemia is a leading cause of nosocomial bloodstream infections, associated with high mortality and substantial healthcare costs. Management challenges primarily arise from diagnostic delays due to the limitations of conventional methods, along with the difficulty in optimizing antifungal regimens. This review provides a comprehensive analysis of these issues, beginning with an overview of the epidemiology, species distribution, and origins of candidemia. It then focuses on the need for improved diagnostic technologies and the importance of susceptibility-guided therapy in enhancing patient outcomes and combating resistance.

The clinical significance of C. auris is undeniable, given its rapid global spread, its resistance to multiple classes of antifungals, and its capacity to cause invasive infections associated with high mortality rates, which highlights the urgent need for rapid and reliable diagnostic methods to identify colonized patients and implement timely infection control measures. Screening for C. auris using culture-based methodologies—relying on morphological, biochemical, and phenotypic characteristics of isolates—remains challenging due to the inherently slow growth of the organism and the limitations of chromogenic media and commercial identification systems in accurately differentiating C. auris from closely related species. In contrast, the Simplexa C. auris Direct assay offers a rapid, accurate, and easy-to-use molecular option, representing a promising advancement in the global effort to combat this emerging pathogen.

Canine parvovirus type 2 (CPV-2) is a highly contagious pathogen causing severe gastrointestinal and cardiac disease in dogs, with three circulating antigenic variants. While quantitative polymerase chain reaction (qPCR) provides accurate diagnosis, its requirement for thermal cycling equipment and trained personnel limits applicability in field settings. This article presents a novel CRISPR-Cas12a based point-of-care diagnostic assay for CPV-2 detection in fecal samples, addressing this diagnostic gap. The assay combines isothermal amplification—either recombinase polymerase amplification (RPA) or loop-mediated isothermal amplification (LAMP)—with Cas12a-mediated detection, eliminating the need for thermal cycling. The CRISPR-Cas12a system recognizes target sequences within the conserved VP2 gene using a guide RNA and activates non-specific nuclease activity, triggering fluorescent signal generation via a reporter probe. This three-step workflow enables rapid, sensitive detection suitable for resource-limited veterinary settings, improving diagnostic capacity for timely quarantine and therapeutic interventions.

Isothermal nucleic acid amplification technologies (INAATs) represent a significant advancement in infectious disease diagnostics, addressing critical limitations of conventional PCR by eliminating the need for thermal cycling equipment and specialized laboratory infrastructure. These methods—including LAMP, RPA, RAA, MIRA, HDA, RCA, SDA, and NASBA—enable rapid pathogen detection at constant temperature while maintaining high sensitivity and specificity for bacterial, viral, fungal, and parasitic agents. Recent innovations in enzyme engineering, assay design, CRISPR-based detection, biosensors, and microfluidics have substantially enhanced these technologies' performance capabilities. This research initiative seeks to consolidate advances in isothermal amplification across methodological innovations, enzyme development, and diverse applications spanning clinical, veterinary, environmental, and food safety sectors. By integrating isothermal amplification with emerging detection platforms and portable devices, these technologies facilitate the transition toward accessible point-of-care diagnostics, particularly benefiting resource-limited settings where infectious disease burden remains greatest. This comprehensive overview welcomes original research and reviews on next-generation diagnostic platforms combining isothermal amplification with advanced detection methodologies.

Candidemia represents a significant nosocomial bloodstream infection challenge, characterized by high mortality rates and considerable healthcare expenditure. This comprehensive review addresses critical management obstacles, primarily diagnostic delays stemming from conventional method limitations and complexities in optimizing antifungal therapy. The analysis encompasses candidemia epidemiology, species distribution patterns, and infection origins before examining diagnostic advancements and the critical role of susceptibility-guided therapy in improving patient outcomes and mitigating resistance development. The review evaluates current diagnostic approaches alongside emerging technologies designed to enhance detection performance. Therapeutic strategies receive detailed scrutiny, including empiric and targeted regimens, mixed infection management, and host-drug interactions, with particular attention to antifungal resistance and combination therapy potential. Recent drug development advances are highlighted, featuring agents in clinical trials targeting novel fungal pathways. Future directions encompassing artificial intelligence integration in diagnostics, vaccine-based prophylaxis approaches, and synergistic treatment strategies are explored. By synthesizing contemporary progress, this review provides clinicians and researchers with evidence-based guidance for navigating candidemia management's evolving landscape.

Antibiotic resistance poses a critical global health challenge requiring advanced investigation methods. Researchers have developed a microfluidic hollow-fiber infection model that simulates realistic pharmacokinetic and pharmacodynamic conditions to study bacterial responses to antibiotic treatment. The system combines dynamic antibiotic concentration gradients with high-resolution imaging of bacteria within tissue-like environments, utilizing minimal resources. Testing with Escherichia coli strains revealed that treatment efficacy depends not only on time above minimum inhibitory concentration, but critically on dosing intervals and recovery phases between doses. Prolonged dosing periods and shortened recovery times enhance bacterial clearance, while specific regimens trigger distinct phenotypic responses including filamentous growth. This innovative platform enables mechanistic analysis of how dosing schedules influence bacterial survival and adaptation, offering valuable insights for optimizing antibiotic treatment strategies and translating findings from laboratory to clinical practice.

# Summary Clinical microbiology of fungal infections represents a critical area of medical laboratory science addressing the diagnosis, identification, and characterization of pathogenic fungi. This comprehensive resource examines diagnostic methodologies, including microscopy, culture techniques, and molecular approaches essential for accurate fungal identification. The content encompasses various clinically significant fungal pathogens, their epidemiology, and associated disease manifestations across diverse patient populations. Understanding fungal infection microbiology is vital for clinicians and laboratory professionals to implement appropriate diagnostic protocols, ensure timely treatment initiation, and improve patient outcomes. The field continues evolving with advancing technologies in fungal detection and antifungal susceptibility testing.

QuickMIC® represents a transformative breakthrough in sepsis management, addressing the critical time constraints inherent in treating this life-threatening condition. By delivering antimicrobial susceptibility testing results in hours rather than the traditional 24-48 hours, QuickMIC® enables clinicians to make informed antibiotic treatment decisions significantly faster. This expedited diagnostic capability enhances patient outcomes by facilitating precise antibiotic selection, thereby reducing the risk of treatment failure and antimicrobial resistance. Recognized as an FDA breakthrough device, QuickMIC® is already operational in European healthcare facilities and is progressing through clinical studies for North American deployment. This innovative technology promises to substantially improve sepsis management protocols and patient survival rates across global healthcare systems.