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BACKGROUND: This study characterizes the clinical utility and validity of the Karius test (KT), a plasma microbial cell-free DNA sequencing platform, as an infection surveillance tool among hematopoietic stem cell transplant (HCT) recipients, including monitoring for cytomegalovirus (CMV) and detecting infections relative to standard microbiologic testing (SMT). METHODS: A prospective, observational cohort study was performed among adult HCT recipients as inpatients and outpatients. Serial KTs were performed starting with 1 sample within 14 days before HCT, then weekly from 7-63 days posttransplant then monthly from 3-12 months post-HCT. Diagnostic performance of KT versus CMV polymerase chain reaction was evaluated with positive percent agreement and negative percent agreement. Infectious events (

Digital PCR (dPCR) holds immense potential for precisely detecting nucleic acid markers essential for personalized medicine. However, its broader application is hindered by high consumable costs, complex procedures, and restricted multiplexing capabilities. To address these challenges, an all-in-one dPCR system is introduced that eliminates the need for microfabricated chips, offering fully automated operations and enhanced multiplexing capabilities. Using this innovative oscillation-induced droplet generation technique, OsciDrop, this system supports a comprehensive dPCR workflow, including precise liquid handling, pipette-based droplet printing, in situ thermocycling, multicolor fluorescence imaging, and machine learning-driven analysis. The system's reliability is demonstrated by quantifying reference materials and evaluating HER2 copy number variation in breast cancer. Its multiplexing capability is showcased with a quadruplex dPCR assay that detects key EGFR mutations, including 19Del, L858R, and T790M in lung cancer. Moreover, the digital stepwise melting analysis (dSMA) technique is introduced, enabling high-multiplex profiling of seven major EGFR variants spanning 35 subtypes. This innovative dPCR system presents a cost-effective and versatile alternative, overcoming existing limitations and paving the way for transformative advances in precision diagnostics.

We herein present a multifunctional self-priming hairpin probe-based isothermal amplification, termed MSH, enabling one-pot detection of target nucleic acids. The sophisticatedly designed multifunctional self-priming hairpin (MSH) probe recognizes the target and rearranges to prime itself, triggering the amplification reaction powered by the continuously repeated extension, nicking, and target recycling. As a consequence, a large number of double-stranded DNA (dsDNA) amplicons are produced that could be monitored in real-time using a dsDNA-intercalating dye. Based on this unique design approach, the nucleocapsid (N) and the open reading frame 1 ab (ORF1ab) genes of SARS-CoV-2 were successfully detected down to 1.664 fM and 0.770 fM, respectively. The practical applicability of our method was validated by accurately diagnosing 60 clinical samples with 93.33% sensitivity and 96.67% specificity. This isothermal one-pot MSH technique holds great promise as a point-of-care testing protocol for the reliable detection of a wide spectrum of pathogens, particularly in resource-limited settings.

Cutaneous mixed infections involving multiple pathogens present significant diagnostic and therapeutic challenges. Here, we report the first documented case of concurrent cutaneous infection caused by Cryptococcus uniguttulatus and Mycobacterium tuberculosis in a 43-year-old male. Initial histopathological examination revealed fungal organisms, and subsequent culture identified C. uniguttulatus as the causative pathogen. Despite treatment with oral itraconazole and amphotericin B, complete resolution was not achieved. Further molecular diagnostic testing using real-time PCR and next-generation sequencing (NGS) revealed a concurrent M. tuberculosis infection. Following successful management of the cryptococcal infection, combination anti-M. tuberculosis therapy led to significant improvement of the cutaneous lesions. This case represents not only the first reported instance of cutaneous C. uniguttulatus infection but also the first documented case of mixed cutaneous infection involving both Cryptococcus and Mycobacterium species. This case underscores the importance of comprehensive diagnostic approaches, particularly molecular methods, in identifying complex infections that may be missed by conventional culture techniques. Additionally, it highlights the necessity of considering mixed infections in cases refractory to initial therapy, even in immunocompetent individuals. [ABSTRACT FROM AUTHOR]

Pediatric acute myeloid leukemia (AML) is a heterogeneous and aggressive hematological malignancy. Despite advances in treatment, the survival rates remain unsatisfactory, emphasizing the need for innovative therapeutic approaches. This narrative review presents a comprehensive overview of the current approach and likely future directions for pediatric AML. The distinct genetic, epigenetic, and molecular features of pediatric AML contribute to its complex pathophysiology and impact on prognosis. Current treatment practices involve a multifaceted approach combining chemotherapy, molecularly targeted therapies, and hematopoietic stem cell transplantation. However, intensive treatment often leads to significant acute and long-term toxicity. Emerging strategies, including precision medicine, immunotherapy, and novel agents, hold promise for improving outcomes and minimizing adverse effects. Ongoing clinical trials are investigating the potential of these innovative approaches to transform pediatric AML care. By highlighting the evolving treatment paradigms and future perspectives, this review underscores the importance of continued research and development in pediatric AML to enhance the survival rates and quality of life of these young patients. [ABSTRACT FROM AUTHOR]

Purpose: Alternative, noninvasive, cost-effective methods to complement or serve as substitutes to current standard-of-care (SOC) procedures in nonmuscle-invasive bladder cancer (NMIBC) follow-up are needed. Uromonitor is a urine biomarker test detecting bladder cancer recurrence through the screening of TERT , FGFR3 , and KRAS hotspot mutations. The aim of this study was to assess Uromonitor performance by comparing it with the current SOC methods. Materials and Methods: Four hundred thirty-nine patients with 528 NMIBC surveillances were enrolled in this study. All patients underwent SOC methods and provided a urine sample for Uromonitor analysis before undergoing cystoscopy. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated for recurrence and compared with the gold standard cystoscopy plus transurethral resection of bladder tumor histopathology. Results: Uromonitor displayed a sensitivity of 87% (95% CI, 74-95), with only 6 of 47 recurrences failing to be detected; specificity of 99% (98-100); PPV of 93% (82-98); and an NPV of 99% (97-99). Cystoscopy showed a total of 22 false positives (32%) not confirmed by transurethral resection of bladder tumor, whereas Uromonitor presented only 3 positive tests where no lesions were found. Overall recurrence rate was 8.9% (n = 47) among 528 total screenings. Sensitivity, specificity, PPV, and NPV values for Uromonitor remained high across all NMIBC grades and stages. Conclusions: Uromonitor represents a reliable tool in the detection of NMIBC recurrence in patients undergoing routine surveillance, regardless of stage and grade. To our knowledge, this is the largest single-center study assessing Uromonitor's performance, thus validating its usefulness in clinical practice. [ABSTRACT FROM AUTHOR]

Encephalitis, a severe brain inflammation, can arise due to various infectious agents, including viruses like Parvovirus B19 (B19V). Previously linked to mild neonatal and young one's illnesses and some haematological diseases, recent evidence associates B19V with encephalitis, with no clear prevalence and mechanisms in place. This systematic review and meta-analysis aim to determine the prevalence of B19V in cases of encephalitis, exploring variations associated with diagnostic approaches, and identifying gaps in existing research to enhance clinical comprehension and diagnostic methods. An extensive search (1994–2024) was performed through PubMed, Scopus, ScienceDirect, and Cochrane databases for research and epidemiological investigations related to B19V in cases of encephalitis. Inclusion criteria focused on studies that verified B19V using molecular (PCR, NGS) or serological (IgM/IgG) techniques in cerebrospinal fluid or serum. Data analysis was done to pool the prevalence data of included studies using a random-effects model. Heterogeneity was evaluated using I2 statistics. Sensitivity and meta-regression analyses were conducted to evaluate variability and the effects of moderators. A total of fourteen studies involving 3,135 encephalitis patients resulted in a combined prevalence of 3% (95% CI: 2–4%). Studies using PCR indicated a greater prevalence (3%) in comparison to ELISA (1%) and NGS (2%). A moderate level of heterogeneity (I2 = 57.4%) was attributed to the variability in diagnostic methods and geographic distribution. Sensitivity analyses validated strong estimates, while meta-regression revealed country as a key moderator accounting for heterogeneity. Publication bias was modest. The research indicates that B19V may be involved in certain encephalitis instances, with an overall prevalence of 3%. The differences observed in the studies emphasize the need for standardized diagnostic procedures and more extensive multicentric epidemiological research. [ABSTRACT FROM AUTHOR]

Purpose of Review: This review aims to summarize current knowledge on the transmission, diagnostics, treatment, and control of sporotrichosis, a global mycosis affecting humans and cats, with a focus on recent taxonomic developments and the rise of cat-transmitted cases. Recent Findings: Sporothrix has undergone considerable taxonomic shifts, with 70 species recognized, each exhibiting diverse biological characteristics and ecological niches, which include pathogens affecting mammals, plants, and insects, as well as fungicolous species. Sporotrichosis is transmitted primarily through the traumatic inoculation of contaminated plant material, but animal-mediated transmission, chiefly via cats infected with highly virulent S. brasiliensis, has increased dramatically. This shift in transmission patterns has led to significant outbreaks, especially in South America, raising concerns about the potential for global dissemination. Advances in PCR-based diagnostics have revolutionized species identification and epidemiological tracking, enabling a deeper understanding of disease transmission dynamics and the genetic diversity of Sporothrix. Genotyping studies have provided insights into the population structure, evolutionary relationships, and potential adaptations of Sporothrix, revealing the complex interplay between human activities, animal interactions, and environmental changes in shaping the distribution of Sporothrix species. Summary: The increasing global incidence of sporotrichosis underscores the need for improved surveillance, better diagnostic accessibility, and more affordable molecular tests. Effective control strategies, including public awareness and responsible animal management, are essential to address the growing global threat of sporotrichosis. [ABSTRACT FROM AUTHOR]

Simple Summary: Endogenous Cushing's syndrome is a rare disorder caused by excessive cortisol most commonly due to elevated ACTH produced by neuroendocrine tumors. These tumors most commonly arise in the pituitary gland (Cushing's disease) but can also originate in other parts of the body (ectopic ACTH syndrome). Distinguishing between pituitary and ectopic causes is essential for choosing the best treatment, yet current tools are not always optimal. Our research compares the transcriptome patterns of pituitary and ectopic tumors to discover unique markers and possible new drug targets. This research provides new insights into specific molecules and pathways linked to each tumor type, common mechanisms of ACTH-secretion, and suggests promising diagnostics and treatment targets for future research. The analysis of the experimental efficacy of existing medications registered for other oncological disorders may provide new perspectives for these rare orphan disorders in cases where traditional treatment options have been exhausted. Background/Objectives: Endogenous Cushing's syndrome (CS) is a rare neuroendocrine disorder often resulting from adrenocorticotropic hormone (ACTH) production by a pituitary or ectopic tumor. The biological mechanisms underlying ACTH-secreting tumors, especially non-pituitary, remain poorly understood. This study aimed to explore the transcriptomic profiles of pituitary and ectopic ACTH-secreting tumors (ASTs) to identify potential biomarkers for differential diagnosis and new therapeutic targets. Methods: In this study, we performed a transcriptome-wide analysis of twenty-pituitary, one pancreatic, and six lung ASTs, along with seven pituitary, eight pancreatic, and eight lung healthy control tissue samples, respectively. Results: The transcriptomic analysis revealed distinct profiles between pituitary and ectopic ACTH-secreting tumors. In all ASTs, except for the pancreatic sample, we found an overexpression of the TBX19 and PITX1 transcription factor genes, which promote the overexpression of POMC. The transcriptional profiles of ectopic lung ASTs showed marked overexpression of the gastrin-releasing peptide and calcitonin genes, which may be potentially useful for diagnostic purposes. We identified several transcriptionally activated molecular pathways in ASTs that might be targetable with existing medications to treat cancer, the most potentially promising of which are RET inhibitors and an IL6R inhibitor. We also predicted and experimentally validated that corticotropinomas express ganglioside 2 molecules, offering new therapeutic targets for Cushing's disease. Conclusions: This study provides new insights into the molecular features of ACTH-secreting tumors and identifies potential diagnostic markers and therapeutic targets. Distinct transcriptomic profiles in pituitary and ectopic tumors highlight unique biomarkers that may improve diagnostic accuracy and facilitate the development of targeted therapies for ACTH-dependent CS. [ABSTRACT FROM AUTHOR]

Aleurocanthus spiniferus (Quaintance) (Hemiptera: Aleyrodidae) has rapidly spread, mainly in the central and eastern Mediterranean coastal area, infesting various new host plants alongside known ones. This invasive species poses a significant threat to agricultural ecosystems, necessitating urgent action to monitor and control outbreaks in previously pest-free areas. While entomological and morphological recognitions are crucial for initial detection, challenges often arise in quickly identifying different developmental stages or genus-level distinctions, particularly in surveys conducted by personnel with limited entomological skills. Due to these challenges, a qPCR probe protocol was developed to enhance the diagnostic capacity of laboratories responsible for the territorial control of pests. This biomolecular tool integrates morphological surveys, enabling prompt and reliable proof of A. spiniferus presence in free areas, delimited territories, or during phytosanitary import inspections. The protocol's high analytical specificity, inclusivity, and exclusivity ensure accurate identification of A. spiniferus, while its low limit of detection and high repeatability and reproducibility reinforce its utility as a standardized diagnostic method. By facilitating prompt and targeted control efforts, this innovative approach strengthens the resilience of agricultural systems against the widespread threat of A. spiniferus infestations. [ABSTRACT FROM AUTHOR]

Simple Summary: Grape phylloxera is a serious insect pest of grapevines worldwide. The past two decades have seen genotypic identification of phylloxera using microsatellite markers become an integral part of control, informing the selection of resistant rootstocks and implementation of quarantine zones to prevent the spread of highly virulent genotypes. Here, we assessed three different molecular methods for screening phylloxera microsatellites, providing comparisons with previous data using newer laboratory approaches, including phylloxera whole-genome DNA sequence data. These comparisons and the standard laboratory protocols presented will allow molecular diagnostic results to be consistently obtained between different research groups and maintain compatibility of future work with valuable historic datasets. Grape phylloxera, Daktulosphaira vitifoliae (Fitch), is an economically significant pest of grapevines. Identification of phylloxera genotypes is an important aspect of management as genotypes differ in virulence and susceptibility to control using resistant rootstocks. Microsatellite markers developed on polyacrylamide gel systems have been the most widely used molecular method for phylloxera genotype identification, but this approach has been superseded by fluorescent capillary-based genotyping. The current study presents new laboratory methods for amplifying a standard set of eight phylloxera microsatellite markers using PCR-incorporated fluorescently labelled primers, genotyped on an ABI capillary platform. Comparison of allele size data scored on (i) polyacrylamide, (ii) capillary, and (iii) high-throughput sequencing (HTS) platforms revealed that the capillary genotyping most closely matched the HTS allele sizes, while alleles of loci originally scored on a polyacrylamide platform differ in size by up to three base pairs, mostly due to the presence of previously uncharacterised DNA sequence indels. Seven common clonal lineages of phylloxera known from Australia are proposed as reference samples for use in calibrating genotyping systems between platforms and laboratories to ensure universal scoring of allele sizes, providing a critical link for accurately matching previous phylloxera genotype studies with current research. [ABSTRACT FROM AUTHOR]

Background: Lung cancer remains the leading cause of cancer mortality globally with EGFR mutations representing a significant driver in advanced non-small cell lung cancer (aNSCLC). The timely detection of these mutations is critical for initiating targeted therapy, yet tissue biopsy limitations often delay treatment. Methods: This multicenter prospective study evaluated the clinical utility of liquid biopsy (LBx) in real-life settings for the early diagnosis of EGFR mutations in patients with suspected aNSCLC. Circulating tumor DNA (ctDNA) was analyzed using the Cobas EGFR Mutation Test and compared to tissue-based next-generation sequencing (NGS). Results: Among 366 aNSCLC patients tested, LBx demonstrated a significantly shorter median turnaround time (TAT) of 3 days compared to 26 days for tissue NGS (p < 0.001) with 100% specificity and 65% sensitivity for EGFR mutation detection. LBx identified actionable EGFR mutations in cases where tissue biopsy was insufficient or unavailable, enabling 43.7% of patients to commence targeted therapy based on ctDNA results prior to biopsy confirmation. Conclusions: These findings highlight the potential of LBx to reduce diagnostic delays and improve access to personalized therapies in a real-world setting. Integrating LBx into routine diagnostic workflows may address current gaps in molecular testing, ensuring timely and precise treatment for aNSCLC patients. [ABSTRACT FROM AUTHOR]

Oak processionary moth (Thaumetopoea processionea) (OPM) Linnaeus, 1758 (Lepidoptera: Notodontidae) is a serious forestry pest and risk to public health in the UK. The economic and environmental cost of chemical pesticides in managing OPM has driven the need for sustainable, strategies which fit into integrated pest management frameworks, including the use of novel biocontrol methods such as conservation biocontrol.Carcelia iliaca Ratzeburg, 1840 (Diptera: Tachinidae), a specific parasitoid of OPM, is currently the main biocontrol agent of the UK OPM population. However, basic information on C. iliaca life history and rates of parasitism are currently lacking, partly driven by the risks OPM pose to human health, making both study and incorporation of biocontrol into management plans difficult.Here, we design and validate a molecular diagnostic assay based on loop‐mediated isothermal amplification (LAMP) to detect C. iliaca from OPM larval tissue samples collected in the field, overcoming the challenges of studying problematic invasive species such as these.To assess assay performance, diagnostic sensitivity, which was 91%, and specificity, which was 75%, are used alongside limit of detection (600 pg). We discuss the wider applications for LAMP as a cost‐effective tool for studying the natural enemies of insect pests which can be used to inform conservation biocontrol management strategies. [ABSTRACT FROM AUTHOR]

Simple Summary: Cancer is a serious problem for dogs and cats, often diagnosed when it is already advanced and more difficult to treat. This article reviews new ways to find cancer earlier in pets, so they have a better chance of successful treatment. These methods include special scanning techniques powered by computer-based image analysis, blood tests that look for cancer markers without surgery, new ways to examine DNA or other molecules from tumors, and even a simple worm-based test to detect changes linked to cancer. By catching the disease sooner, veterinarians can suggest treatments that are more targeted and less invasive, helping pets live longer and with a higher quality of life. Because cancers in animals and humans share many features, these discoveries can guide research for both species. The goal is to make advanced cancer testing available and affordable in more clinics, leading to earlier detection and better outcomes. Ultimately, this work benefits families, pets, and the broader scientific community. Cancer is a leading cause of death among companion animals, with many cases diagnosed at advanced stages when clinical signs have appeared, and prognosis is poor. Emerging diagnostic technologies, including Artificial Intelligence (AI)-enhanced imaging, liquid biopsies, molecular diagnostics, and nematode-based screening, can improve early detection capabilities in veterinary medicine. These tools offer non-invasive or minimally invasive methods to facilitate earlier detection and treatment planning, addressing the limitations of traditional diagnostics, such as radiography and tissue biopsies. Recent advancements in comparative oncology, which leverage the biological similarities between human and companion animal cancers, underscore their translational value in improving outcomes across species. Technological advances in genomics, bioinformatics, and machine learning are driving a shift toward precision medicine, enabling earlier detection, personalized treatments, and monitoring of disease progression. Liquid biopsy testing detects circulating tumor DNA and tumor cells, providing actionable insights into tumor genetics without invasive procedures. Imaging systems enhance diagnostic precision, offering consistent and accurate tumor identification across veterinary practices, while portable innovations like Caenorhabditis elegans-based screening provide accessible options for underserved regions. As these technologies migrate from human medicine to veterinary applications, they are poised to redefine cancer care for companion animals. This review highlights key advancements in diagnostic technologies and their application in veterinary oncology, with a focus on enhancing early detection, accessibility, and precision in cancer care. By fostering the adoption of these innovations, veterinary oncology can achieve a new standard of care, improving outcomes for both animals and humans through the lens of comparative oncology. [ABSTRACT FROM AUTHOR]

Background: Acute myeloid leukemia (AML) complicated by disseminated intravascular coagulation (DIC) poses major diagnostic and therapeutic challenges. While DIC is well documented in acute promyelocytic leukemia, its manifestations in non-APL AML remain underexplored, necessitating precise diagnostic strategies for effective management. Methods: AML patients with overt DIC were analyzed, including morphological, immunophenotypic, cytogenetic, and genetic evaluations. DIC was diagnosed using the ISTH scoring system, and AML subtypes were classified following WHO criteria. Results: Three diagnostic patterns were identified. (1) Acute promyelocytic leukemia: Leukemia characterized by PML::RARa rearrangements, FLT3 co-mutations, and frequent Auer rods and faggot bundles. Immunocytological analysis showed CD34 and HLA-DR negativity. (2) AML with FLT3 and/or NPM1 mutations: A high prevalence of cup-like blasts was found in 70% of cases. FLT3 mutations, often co-occurring with NPM1, dominated, while karyotypes were typically normal. Immunophenotyping revealed strong myeloid marker expression (MPO+, CD13+, and CD33+), with occasional CD34 negativity. (3) AML with monocytic differentiation: Leukemia defined by monoblastic/promonocytic morphology, DNMT3A mutations, and complex karyotypes or 11q23 rearrangements. Immunophenotyping demonstrated a dominance of monocytic markers (CD4+, CD14+, CD15+, and CD64+). Two patients presented unique profiles with no alignment to these patterns. Conclusions: This study highlights distinct hematopathological patterns of AML with overt DIC, providing a framework for early and precise diagnosis. Recognizing these patterns is critical for tailoring diagnostic and therapeutic approaches to improve outcomes in this high-risk population. [ABSTRACT FROM AUTHOR]

Background/Objectives: The accurate detection of EGFR mutations, particularly the rare S768I variant, is crucial for guiding treatment decisions in non-small cell lung cancer (NSCLC) patients. This study investigated the incidence of false positives in S768I mutation detection using the IdyllaTM qPCR system and compared results with next-generation sequencing (NGS). Methods: A prospective observational study was conducted at the Dr. Negrín University Hospital between July 2023 and July 2024. Six NSCLC patient samples with S768I variant detection by IdyllaTM were analyzed from all NSCLC cases tested during the study period. Initial testing was performed on tissue samples (Idylla1), followed by replicate analysis using extracted DNA (Idylla2). Results were compared with NGS as the reference method. Statistical analysis included the calculation of sensitivity, specificity, accuracy, and Kappa concordance index. Results: Initial Idylla testing showed an 80% false positive rate, with only one of five positive results confirmed by NGS. The first analysis demonstrated high sensitivity (100%) but low specificity (20%), with an accuracy of 0.333 and poor concordance with NGS (Kappa = 0.077). Repeat testing using extracted DNA showed improved performance, with increased accuracy (0.833) and better agreement with NGS (Kappa = 0.571). Analysis of amplification curves revealed that false positives typically showed normalized fluorescence values below 12 points, with no clear correlation between false positives and factors such as sample quantity or tumor content. Conclusions: While the IdyllaTM system shows high sensitivity for S768I detection, its initial specificity is problematic, leading to frequent false positives. These findings emphasize the importance of confirming positive S768I results through alternative methods like NGS, particularly when these results could influence therapeutic decisions. Results suggest the need to refine the system's interpretation algorithms to improve specificity. [ABSTRACT FROM AUTHOR]

Introduction: Nosocomial lower respiratory tract infections (nLRTIs), including hospital-acquired pneumonia (HAP) and ventilator-associated pneumonia (VAP), remain significant challenges due to high mortality, morbidity, and healthcare costs. Implementing accurate and timely diagnostic strategies is pivotal for guiding optimized antimicrobial therapy and addressing the growing threat of antimicrobial resistance. Areas Covered: This review examines emerging microbiological diagnostic methods for nLRTIs. Although widely utilized, traditional culture-based techniques are hindered by prolonged processing times, limiting their clinical utility in timely decision-making. Advanced molecular tools, such as real-time PCR and multiplex PCR, allow rapid pathogen identification but are constrained by predefined panels. Metagenomic next-generation sequencing (mNGS) provides comprehensive pathogen detection and resistance profiling yet faces cost, complexity, and interpretation challenges. Non-invasive methods, including exhaled breath analysis using electronic nose (e-nose) technology, gene expression profiling, and biomarker detection, hold promise for rapid and bedside diagnostics but require further validation to establish clinical applicability. Expert Opinion: Integrating molecular, metagenomic, biomarker-associated, and traditional diagnostics is essential for overcoming limitations. Continued technological refinements and cost reductions will enable broader clinical implementation. These innovations promise to enhance diagnostic accuracy, facilitate targeted therapy, and improve patient outcomes while contributing to global efforts to mitigate antimicrobial resistance. [ABSTRACT FROM AUTHOR]

Objective: The clinical diagnosis of respiratory tract infections (RTIs) may result in unnecessary antibiotic treatment due to clinical exams' low sensitivity and specificity to differentiate viral from bacterial infections and costly diagnostic work-ups. Unnecessary antibiotic consumption drives antibiotic resistance. We explored whether a rapid influenza-specific polymerase chain reaction (PCR) assay reduced antibiotic use in an emergency room before the COVID-19 pandemic. Methods: We conducted an observational retrospective study of patients with RTI symptoms treated in the ER of the University Hospital Basel from September 2014 to June 2015. We evaluated the impact of rapid diagnostic results, such as an influenza-specific PCR, blood sample results, and radiological imaging, on antibiotic prescription rates. Patient-related confounding factors were included since a patient's clinical condition affects doctors' clinical decision-making. Results: We included 607 patients with RTIs, tested with PCR for influenza A or B. Logistic regression showed that the odds ratio (OR) of being treated with antibiotics was significantly reduced by more than two-thirds in patients with a positive influenza PCR result (OR = 0.37; 95% CI, 0.22–0.59; p < 0.001). Increasing C-reactive protein (CRP) levels by tenfold (OR = 5.14; 95% CI, 3.34–8.12; p < 0.001) or suspected chest infection on a radiograph (OR = 5.81; 95% CI, 3.23–10.89; p < 0.001) increased the OR of antibiotic treatment by fivefold. The highest OR for antibiotic prescription was due to increased procalcitonin level by tenfold (OR = 10.13; 95% CI, 4.79–23.4; p < 0.001). Conclusions: Our study provides real-world evidence from a pre-COVID-19 ER setting of diagnostic tools used for RTI evaluation and their impact on antibiotic prescriptions. Rapid influenza-specific PCR results may affect the prescription rate of antibiotics but should be seen as part of a comprehensive diagnostic approach to guide clinical decision-making. [ABSTRACT FROM AUTHOR]

Background: High-grade advanced ovarian, fallopian tube, and primary peritoneal (HGAOC) cancers require both surgical and systemic treatment. The introduction of polyADP-ribose polymerase inhibitors (PARPis) has significantly improved outcomes. This study presents an analysis of HGAOC patients treated at a single center, following updated guidelines. Methods: We observed 437 women newly diagnosed with HGAOC at the Department of Gynecological Oncology between January 2018 and December 2023. Results: Since November 2022, first-line treatment has included bevacizumab and PARPi, regardless of residual disease post-cytoreductive surgery. In both BRCA1/2-mutated and non-mutated groups, PARPi-based regimens increased significantly after May 2021 (p < 0.01). Recurrence number emerged as a strong prognostic factor for survival (p < 0.001), with each recurrence raising mortality risk by 80%. Median survival was 21 months for paclitaxel + platinum derivatives (PC), 27 months for PC + bevacizumab (BEV), and 30 months for PC + BEV + PARPi. Conclusions: The rapid adoption of modern therapies at our center has aligned treatment strategies with HRD status and global standards. However, variations in financial regulations and drug accessibility persist across countries. Despite these challenges, physicians should prioritize the most effective therapies available. [ABSTRACT FROM AUTHOR]

Background/Objectives: Candida auris is an emerging multidrug-resistant pathogen with the potential to cause invasive fungal infections and healthcare-associated outbreaks. Currently, there is no systematic review explicitly focusing on the up-to-date molecular diagnostics of this pathogen to cover the entire process, including sample pre-extraction procedures, nucleic acid extraction, and DNA-based detection. Sample pre-treatment and extraction are the prerequisites before molecular testing and have implications on the downstream detection but have not been reviewed elsewhere. This review aims to summarize a comprehensive update in the past 5 years. Methods: A systematic review was conducted to search for articles published in the period between 1 January 2020 and 20 November 2024 from various databases, including PubMed, Google Scholar, and Web of Science. The findings were produced through narrative synthesis, with quantitative analysis conducted where applicable. Results: Starting from 1115 records, 28 studies that met the inclusion criteria were included in the analysis. This review summarized the key updates on three categories, including (i) sample pre-extraction procedures and nucleic acid extraction, including magnetic, bead-beating, mechanical, chemical, thermal, and column-based protocols; (ii) commercial molecular assays; and (iii) laboratory-developed tests (LDTs). For real-time PCR, commercial molecular assays and LDTs showed sensitivity (ranging from 94.9% to 100% and 44% to 100%, respectively) and specificity (ranging from 98.2% to 100% and 92% to 100%, respectively). Conclusions: Here, we describe a useful summary to enlighten readers from clinical microbiology laboratories on the nucleic acid extraction protocols and performance of various molecular diagnostic assays used for the detection of C. auris. [ABSTRACT FROM AUTHOR]

Hearing loss is a prevalent condition with a significant impact on individuals' quality of life. However, comprehensive studies investigating the differential gene expression and regulatory mechanisms associated with hearing loss are lacking, particularly in the context of diverse patient samples. In this study, we integrated data from 10 patients across different regions, age groups, and genders, with their data retrieved from a public transcriptome database, to explore the molecular basis of hearing loss. These samples are mainly from fibroblasts and keratinocytes. Through differential gene expression analysis, we identified key genes, including ICAM1, SLC1A1, and CD24, which have already been shown to play important roles in neurogenic hearing loss. Furthermore, we predicted potential transcriptional regulatory factors that may modulate the expression of these genes. Enrichment analysis revealed biological processes and pathways associated with hearing loss, highlighting the involvement of circadian rhythm disruption and other neuro-related disorders. Although our study is limited by the sample size and the absence of larger-scale investigations, the identified genes and regulatory factors provide valuable insights into the molecular mechanisms underlying hearing loss. Further molecular and cellular experiments are necessary to validate these findings and elucidate the precise regulatory mechanisms involved. In conclusion, our study contributes to the understanding of hearing loss pathogenesis and offers potential targets for molecular diagnostics and gene-based therapies. This provides a foundation for further research into personalized approaches to diagnosing and treating hearing loss. [ABSTRACT FROM AUTHOR]

Purpose: The NAB2::STAT6 fusion is predominantly associated with solitary fibrous tumors (SFTs) and is utilized in diagnosing SFTs through nuclear STAT6 protein overexpression. Recent studies expanded the phenotypic spectrum of NAB2::STAT6 rearranged neoplasms, including adamantinoma-like and teratocarcinosarcoma-like phenotypes. We report a case of a NAB2::STAT6 rearranged epithelial tumor exhibiting sebaceous differentiation in the parotid gland. Methods: Fine-needle aspiration (FNA), histopathology, immunohistochemistry, and molecular studies of this case were described. Results: Fine-needle aspiration (FNA) revealed atypical basaloid cells, suggesting primary salivary gland carcinoma or metastasis. Histological examination showed basaloid-squamous cells with a monomorphic appearance containing foci of sebaceous differentiation, expressing pancytokeratin, p40, and androgen receptor, while CD34 staining was negative. Molecular studies identified a NAB2::STAT6 fusion, along with an AKT1 mutation. Conclusion: This case further expands the phenotypic spectrum of NAB2::STAT6 rearranged neoplasms and emphasizes comprehensive histopathological and molecular analysis in challenging head and neck tumors. It suggests STAT6 immunohistochemistry as a potential screening tool for head and neck tumors resembling sebaceous carcinoma, myoepithelial tumors, or GLI1-altered neoplasms. [ABSTRACT FROM AUTHOR]

The seasonality and epidemiology of viral acute respiratory infections (ARIs) have changed since the coronavirus disease 2019 pandemic. However, molecular-based ARI surveillance has not been conducted in Japan. We developed a regional surveillance program to define the local epidemiology of ARIs. Between December 2023 and March 2024, 2,992 upper respiratory samples collected from patients suspected of having ARIs at five facilities in Kyoto City, Japan, were tested for SARS-CoV-2, influenza virus, and respiratory syncytial virus (RSV) using RT‒PCR. Samples negative for these viruses were randomly selected for testing with the FilmArray Respiratory Panel, and the detection rates of other viruses were estimated. SARS-CoV-2, influenza virus, and RSV were detected in 598 (20.3%), 165 (5.6%), and 40 (1.4%) of the 2,949 samples with valid RT‒PCR results, respectively. The most prevalent viruses in the < 6, 6–17, 18–64, and ≥ 65 year age groups were rhinovirus/enterovirus, RSV, and SARS-CoV-2; influenza virus, seasonal coronavirus, and rhinovirus/enterovirus; SARS-CoV-2, seasonal coronavirus, and influenza virus; and SARS-CoV-2, seasonal coronavirus, and influenza virus, respectively. Significant differences in the detection rates of these viruses were detected between the age groups. This study highlights the importance of age-stratified molecular-based surveillance for a comprehensive understanding of the epidemiology of ARIs. [ABSTRACT FROM AUTHOR]

Quantitative polymerase chain reaction (qPCR) and digital PCR (dPCR) are applied for quantifying molecular targets in disease diagnostics, pathogen detection, and ecological monitoring. Uptake of dPCR is increasing due to its higher quantification accuracy relative to qPCR, which stems from its independence from standard curves and its increased resistance to PCR inhibitors. Throughput can be increased through multiplexing, which allows simultaneous quantification of multiple targets. However, multiplexing with dPCR faces unique challenges relative to qPCR. Here, we describe the three-phase development process of non-competing multiplex dPCR assays using target-specific fluorescently labeled hydrolysis probes. We highlight common challenges encountered, along with recommended solutions. Phase 1: In silico assay design; target-specific primers and probes are selected or designed, potential issues with primer and probe interactions are identified, and fluorophores and quenchers are chosen based on dPCR instrumentation. Phase 2: Wet-lab validation; assays are benchmarked using positive controls. Insufficient performance leads to assay redesign, as needed. Phase 3: Assay implementation; assay specificity and sensitivity are validated on relevant sample matrices. Finally, we provide recommendations on the future design and standardization of multiplexed dPCR assays, highlighting the need for better in silico predictions of assay performance, standardizing positive controls, and automating partition classification systems. [ABSTRACT FROM AUTHOR]

Background/Objectives: This study explores the potential of using elevated levels of blood double-stranded RNA (dsRNA) as a diagnostic tool for severe fever with thrombocytopenia syndrome (SFTS) infection. Methods: Blood samples from SFTS patients were collected, dsRNA was purified, and total dsRNA expression was quantitatively analyzed using a spiropyran-based method. Comparative analysis was performed using blood samples from healthy individuals and scrub typhus patients with similar symptoms. Results: The results revealed that individuals infected with SFTS had significantly higher total blood dsRNA levels compared to healthy or scrub typhus controls. The dsRNA-based method also has potential for assessing infection severity based on dsRNA levels. Conclusions: These findings suggest that total dsRNA expression can serve as a quick and convenient method to differentiate SFTS from other non-viral conditions with similar clinical presentations. This method shows promise as a novel diagnostic tool. [ABSTRACT FROM AUTHOR]

The rapid, precise, and automated diagnosis of infectious diseases is crucial for effective disease management and control. Herein, the integrated portable and automatic digital detection system (IPADS), a novel diagnostic platform for nucleic acid detection is introduced. The device employs the hybrid magnetic system (HMS), which uses an electromagnet and a movable permanent magnet to modulate the magnetic field and control bead movement, increasing nucleic acid extraction efficiency to over 80%, while simplifying the traditional labor‐intensive process and enabling quick, low‐risk sample processing. Additionally, a disposable cartridge is designed for integrated HMS based preprocessing, with detection performed using digital RPA‐Cas12a, enabling rapid, enclosed, and automation‐friendly detection across a dynamic range spanning five orders of magnitude, with a sensitivity as low as 100 copies mL−1 in serum samples. An automated platform further optimizes workflow. As a proof of concept, IPADS is applied to detect hepatitis B virus (HBV) DNA in 20 clinical serum samples, demonstrating high concordance with gold‐standard quantitative PCR (qPCR) methods. These results validate the potential of IPADS as a reliable point‐of‐care testing solution. [ABSTRACT FROM AUTHOR]

Fungal infections are common in highly immunosuppressed, solid organ transplant recipients. They can be quite difficult to diagnose in a timely manner; thus, we present a review of current studies focusing on broad categories of molecular diagnostics, i.e., metagenomic sequencing, magnetic resonance, and gas chromatography mass spectrometry. We further discuss their syndrome-specific utilization in the diagnosis of fungemia and disseminated disease, pneumonia, and central nervous system infections. We assess the level of evidence of their utility as fungal diagnostics particularly in solid organ transplant recipients using the STARD criteria. In addition, we provide future research directions to substantiate and appropriately utilize these platforms in clinical practice. Directed polymerase chain reaction testing and targeted metagenomic sequencing are being used clinically and show the most promise, though only in conjunction with conventional methods at this time. The majority of these platforms contain limited data, and thus further larger studies are needed in order to properly implement their use. [ABSTRACT FROM AUTHOR]

Introduction: Chlamydia trachomatis continues to be the most common bacterial infection worldwide and rates continue to increase despite long-standing control efforts. Point of care (POC) testing options may offer improvements in case finding that lead to improved control of this sexually transmitted infection (STI). Areas covered: This review will provide information on the three tests that have US Food and Drug Administration (FDA) clearance and describe assays in the developmental pipeline. The review will also provide implementation evaluations of the existing tests and offer suggestions about factors to consider prior to adoption of these or newer tests as they become available. Expert opinion: Technology is developing rapidly and may soon offer many choices of rapid diagnostic tools which can be used in clinical settings to detect chlamydial infections, particularly in underserved populations. The key to successful deployment of new tests will rest on data generated by implementation research to identify the features that create barriers or facilitate adoption of a new clinical paradigm. [ABSTRACT FROM AUTHOR]

We present a cost-effective and simple multiplex nucleic acid quantification method using droplet digital PCR (ddPCR) with digital melting curve analysis (MCA). This approach eliminates the need for complex fluorescent probe design, reducing both costs and dependence on fluorescence channels. We developed a convolutional neighborhood search algorithm to correct droplet displacement during heating, ensuring precise tracking and accurate extraction of melting curves. An experimental protocol for digital MCA on the ddPCR platform was established, enabling accurate quantification of six target pathogen genes using a single fluorescence channel, with an average accuracy of 85%. Our method overcomes the multiplexing limitations of ddPCR, facilitating its application in multi-target pathogen detection. [ABSTRACT FROM AUTHOR]