Journal of Clinical Virology

Hybrid capture approaches can detect a broad range of pathogens directly from clinical samples with greater sensitivity than untargeted approaches. Clinical adoption has been limited by long turnaround times, complex workflows, and perceived high costs. We iteratively developed and validated a rapid hybrid capture workflow based on Illumina's RPIP protocol (standard workflow). Rapid-RPIP v1 reduced wet-lab time from ~11.4 to ~7.3 h, for a turnaround time (TAT) of <15 h. Rapid-RPIP v2 further reduced wet-lab time by ~0.7 h and used Illumina Turbo Sequencing shortening the sequencing time from ~5.5 to ~2.5 h thus achieving a TAT of <10 h without compromising sensitivity and yielding higher RPKMs for RNA viruses than the standard workflow. This approach could be readily adapted to a broad range of clinical and public health applications.

Background: Amplicon-based next-generation sequencing is a cornerstone of viral genomic surveillance, but it often produces datasets with highly uneven read coverage due to PCR amplification bias. This leads to excessive data generation and substantial computational overhead for downstream bioinformatic analysis. Results: We evaluated the efficacy of two data reduction strategies on 220 SARS-CoV-2 clinical specimens and 12 wastewater samples: random subsampling with SeqKit and read depth normalization with BBNorm. FASTQ files were reduced to target depths from 10X to 10,000X to compare genome coverage, lineage assignment concordance, and computational resource usage. Both strategies markedly reduced total processing time. For clinical isolates, read depth normalization corrected for amplification bias resulting in highly concordant lineage assignments at a target depth of just 200X. Random subsampling preserved inherent bias but produced reliable assignments at depths over 500X. Reducing target depths below 500X in complex wastewater mixtures severely degraded variant deconvolution accuracy and heavily skewed lineage abundance profiles for both methods. Conclusion: Both random subsampling and read depth normalization successfully reduce computational overhead while preserving key clinical sequencing metrics at appropriate reduction settings. The optimal method depends on the analytical goal, ranging from computationally lightweight random subsampling for rapid lineage assignment, to read depth normalization for generating high-quality consensus sequences. However, aggressive data reduction should be avoided in environmental surveillance, where maintaining a high baseline sequencing depth is critical for accurate mixed-variant deconvolution.

In resource-limited settings plasma is usually transported to centralized laboratories for HIV RNA quantification; often by motor vehicles using dry ice. Any decay in plasma HIV RNA when near the threshold used to define virologic failure could impact clinical management. However, maintaining a cold chain during transport can be costly and limit the frequency of shipments, which can delay clinical assessments and also negatively impact patient care. Transporting plasma at ambient temperatures could facilitate timely measurements of plasma HIV RNA viral loads (VL). We conducted a stability study of HIV RNA in plasma held for 1, 3, 5, and 7 days at a range of temperatures (4{degrees}C, 30{degrees}C, and 37{degrees}C) to simulate the durations and temperatures specimens could experience during transport. Thirteen large volume plasma specimens with VL ranging from 2.7-5.53 log10 copies/mL were evaluated. Mean changes in VL between Day-0 to Day-1, Day-3, Day-5 or Day-7 were calculated, with decays >0.5 log10 c/mL considered clinically relevant. Compared to Day 0, the VL of plasma stored at 4{degrees}C and 30{degrees}C remained stable across the Days tested. The VL in plasma stored at 37{degrees}C had a progressive decline at Day-1, Day-3, Day-5 and Day-7 (means of -0.04, -0.19, -0.33. -0.34 log10 c/mL, respectively), with a decay of >0.5 log10 observed in 3/13 (23%) specimens at Day-7. A linear regression analysis showed that plasma specimens stored at 37{degrees}C significantly decayed over time (slope -0.054, 95% CI -0.079, -0.029; p=0.01). Any decay observed among specimens stored at 4{degrees}C and 30{degrees}C was not clinically relevant nor statistically significant. Our observations suggest that HIV RNA levels in plasma should remain stable if transported to centralized laboratories at ambient temperatures of [&le;]30{degrees}C or chilled by cold packs over a period [&le;]7 days. Transporting specimens with cold packs instead of dry ice could reduce costs and facilitate the timely monitoring of VL used to assess and guide antiretroviral therapy.

Healthcare workers are occupationally exposed to blood and body fluids and require effective protection against hepatitis B virus infection. Evidence on hepatitis B surface antibody (anti-HBs) seroprotection among healthcare workers in Namibia remains limited. We estimated the prevalence of anti-HBs seroprotection and examined associated factors among healthcare workers tested at Eastgate Clinic in Windhoek, Namibia. We conducted a cross-sectional analysis of routinely collected anti-HBs test records from 1 January to 31 December 2025. The first chronological test per individual was included. Anti-HBs concentrations [&ge;]10 mIU/mL were classified as seroprotective. Results outside the assay reporting range were treated as censored but remained classifiable for the binary outcome. Prevalence estimates were reported with Wilson 95% confidence intervals (CIs). Modified Poisson regression with robust standard errors was used to estimate adjusted prevalence ratios (aPRs). The register contained 304 test records representing 299 unique individuals. After excluding three individuals with missing anti-HBs results, 296 were analysed; 163 (55.1%) were female. Overall, 148 individuals were seroprotected, corresponding to a prevalence of 50.0% (95% CI 44.3-55.7%). Seroprotection was lowest among participants aged 18-29 years (35.3%) and highest among those aged 50-59 years (71.1%). Each 10-year increase in age was associated with a higher prevalence of seroprotection after adjustment for sex (aPR 1.14, 95% CI 1.06-1.24; p=0.001), whereas sex was not independently associated with seroprotection (aPR 0.97, 95% CI 0.78-1.22; p=0.797). The age association remained consistent across sensitivity analyses. Half of the healthcare workers tested had anti-HBs concentrations below the operational seroprotection threshold. These findings support systematic vaccination verification, appropriately timed post-vaccination testing, and documented follow-up of non-protective results. The absence of vaccination history, occupational characteristics, testing indications, and additional hepatitis B serological markers limited causal and clinical interpretation.

BackgroundHuman Mastadenovirus B (HAdV-B) causes ocular and respiratory infections and is capable of causing outbreaks in healthcare settings. HAdV-B114, a recombinant genotype only formally recognised in 2023, has been reported predominantly from Germany, the USA, China and Japan; genomic data from UK clinical settings remain limited. This study analysed 68 HAdV-B whole genomes from ocular and respiratory-associated samples collected at a Nottingham hospital across the pre- and post-COVID-19 restriction periods (2017-2023). MethodsClinical samples typed positive for HAdV-B by partial hexon Sanger sequencing were selected for whole-genome sequencing using a novel overlapping PCR amplicon strategy, sequenced on the Oxford Nanopore Technologies PromethION platform. Phylogenetic and pairwise difference analyses were used to characterise genotype composition and genomic relatedness. SimPlot and amino-acid comparison characterised divergence between sub-lineages. ResultsWhole-genome phylogenetics confirmed that two species B genotypes were detected in Nottingham University Hospitals between 2017-23: HAdV-B114 (n=56) and HAdV-B7 (n=12). HAdV-B114 was detected before and after COVID-19 restrictions, with an increase in 2022-2023, consistent with post-restriction expansion of a pre-existing lineage. Pairwise difference analysis identified 24 genomically identical B114 sequences; associated metadata argued against a single nosocomial transmission chain interpretation, with the pattern more consistent with repeated detection of a widely circulating community strain, with the possibility of localised onward transmission. Within the B114 clade, multiple monophyletic groups were identified by whole-genome phylogenetics. ConclusionsThis study adds whole-genome resolution to the existing UK HAdV-B114 data, demonstrates the utility of WGS for clinical adenovirus surveillance and shows the benefits of combining epidemiological data with genomic analysis.

Macrolides remain the first-line antimicrobial treatment for pertussis, primarily caused by Bordetella pertussis. Emergence of macrolide-resistant B. pertussis in Australia and globally presents a growing concern for clinical management. As molecular diagnosis predominates for pertussis, limited numbers of cultures are available for antimicrobial susceptibility testing. The A2037G mutation in the 23S rRNA gene is most commonly associated with macrolide resistance in B. pertussis, yet no commercial molecular assay is available for its detection. In this study, we adapted and validated a previously published real-time PCR assay as a laboratory developed test (LDT) on the Panther Fusion(R) system using Open Access functionality to detect the macrolide resistance-associated mutation in B. pertussis from culture isolates and clinical specimens. Total turnaround time was ~2.5 h for the first result and 2-3 min for subsequent results in the same batch, with an estimated cost of AUD 18 per sample, excluding labour. A total of 30 samples were included in the validation panel to assess assay performance. All isolates and clinical specimens in the validation panel produced results concordant with the expected mutation status, demonstrating 100% accuracy, sensitivity and specificity. The assay demonstrated a limit of detection of 1.5 x 103 CFU/mL (22 CFU/reaction). This Panther Fusion(R) LDT provides a rapid and fully automated approach for molecular detection of the targeted mutation associated with macrolide resistance in B. pertussis. Its integration into routine molecular workflows as a reflex test following identification of B. pertussis may facilitate timely identification of resistant infections.

Broad-range RT-qPCR assays are essential for detecting highly diverse viruses within virus families, but designing suitable primers remains challenging because of extensive sequence diversity and biases in public sequence databases. Here, we present a semi-automated workflow that combines sequence clustering and varVAMP to streamline pan-RT-qPCR primer design. The workflow reduces sequence redundancy, improves the representation of viral diversity, and facilitates the identification of broadly reactive primer candidates. We demonstrate the workflow using the arboviral genera Orthoflavivirus and Alphavirus as proof-of-concept datasets, showing its applicability to large and genetically diverse viral sequence collections. The workflow provides a practical, scalable, and reproducible framework for developing broad-range primer candidates and may support surveillance, diagnostics, and preparedness for emerging or divergent viruses.

Background: Filoviruses, specifically Ebola virus (EBOV), Bundibugyo virus (BDBV), and Marburg virus (MARV), have been the etiologic agents of multiple outbreaks of severe disease in the Democratic Republic of the Congo (DRC). Two EBOV-specific vaccine regimens have been approved for use in outbreak scenarios, including the Ad26.ZEBOV/MVA-BN-Filo (Zabdeno/Mvabea) two-dose vaccine, where the MVA-BN-Filo booster dose contains EBOV, MARV, and Sudan (SUDV) virus glycoproteins (GPs) and the nucleoprotein from Tai Forest virus. While long-term immunogenicity of up to two years has been found in clinical trial settings, limited real-world data exists. Methods: Using a multiplexed immunoassay, samples (n=698 at baseline) collected in Mbandaka, Equateur province, DRC, between September 2023 and October 2024 were assessed for seroreactivity to six filovirus antigens (1 BDBV, 2 EBOV, 2 MARV, and 1 SUDV) at three distinct periods: baseline/pre-vaccination, 6-8 weeks post-first dose (Zabdeno), at booster administration (Mvabea), and one-year post-boost dose. Results: Substantial filovirus seroreactivity was found at baseline, with 48.7% (n=340/698) of individuals being seroreactive to at least one virus antigen. High baseline seroreactivity was found against BDBV GP (31.1%, n=217/698), with lower baseline seroreactivity to EBOV GP (15.5%, n=106/698), comparatively. Overall EBOV GP seroreactivity increased to 66.8% (n=376/563) at one year-post vaccine boost and 57.6% (n=250/434) of participants were seroreactive. Despite the booster dose inclusion of SUDV GP and MARV GP, there were no substantial increases in reactivity against either antigen at subsequent timepoints. Overall filovirus seroreactivity increased at all timepoints for the entire cohort, although filovirus reactivity decreased over the study period for those who were seroreactive at baseline. Findings: These data highlight detection of significantly elevated seoreactivity to EBOV GP following first-dose administration of Ad26.ZEBOV, which remained relatively stable one-year post-booster dose administration. Additionally, this study identifies high levels of baseline seroreactivity to BDBV GP despite no previous documented outbreaks in this region of DRC. These data suggest that vaccination with the Ad26.ZEBOV-MVA-BN-Filo vaccine induces stable seroreactivity against EBOV GP, but not to other filovirus antigens.

Bordetella pertussis is the main aetiological agent of pertussis (whooping cough), with a global resurgence of cases reported in 2024. As current pertussis diagnostics are predominantly based on molecular methods, cultured isolates are increasingly scarce, limiting traditional culture-based surveillance. To address these challenges, this study evaluated culture-independent approaches for B. pertussis surveillance and their feasibility for implementation in public health laboratory settings in Victoria, Australia. The Microbiological Diagnostic Unit Public Health Laboratory received 356 nasopharyngeal clinical specimens that were PCR-positive for Bordetella spp. from diagnostic laboratories for further characterisation during the 2024-2025 epidemic period. Culture-based approaches yielded only five B. pertussis isolates, largely due to limitations associated with specimen transport media. Four culture-independent approaches were evaluated for B. pertussis characterisation: real-time PCR assay for high-resolution Bordetella spp. differentiation (n=356), multiplex PCR amplicon sequencing assay for lineage typing (n=38), in-house laboratory developed test implemented on Panther Fusion(R) System for macrolide resistance screening (n=92) and shotgun metagenomic sequencing (n=5). The dataset was dominated by B. pertussis (330/356; 92.7%) with most strains harbouring ptxP3 (35/38; 92.1%) allele and the macrolide resistance rate was reported as 4.3% (4/92). Shotgun metagenomics was able to detect B. pertussis directly from clinical samples, but the genomic recovery was limited. Evaluation of operational performance and workflow integration highlighted method-specific strengths and limitations, informing suitability for implementation in public health laboratories. This study demonstrates the potential to integrate culture-independent approaches into public health laboratory settings to strengthen B. pertussis surveillance and inform feasible workflow design amid limited culture availability.

Mosquito-borne orthoflaviviruses pose a significant threat to human and animal health worldwide. In Europe, West Nile (WNV), Usutu (USUV) and Bagaza (BAGV) viruses have expanded their geographic distribution and are detected with increasing frequency. Following the emergence of BAGV in 2021, we investigated the circulation dynamics and genomic diversity of orthoflaviviruses in southern Portugal, where their epidemiology remains poorly understood. Active Orthoflavivirus surveillance was conducted in birds and mosquitoes during 2021-2023 using serological and molecular assays, complemented by viral genome sequencing and phylogenomic analyses. Viral RNA was detected in 3.5% (41/1,171) of birds, while seroneutralization assays identified an overall seroprevalence of 55.3% (503/910). WNV RNA (lineage-1, strains WMed-1.2 and WMed-1.3) was detected annually in resident birds, including Alectoris rufa, Cyanopica cooki and Garrulus glandarius (1.5%, 18/1,171), and in Culex univittatus mosquitoes (0.8%, 2/255). Together with a WNV-specific seroprevalence of 28.5% (259/910) in birds, these findings support sustained local circulation. USUV RNA (Africa 3.1 sub-lineage) was detected in A. rufa in 2021 and 2023 (0.2%, 2/1,171), but seroprevalence was low (1.3%, 8/623), consistent with intermittent circulation. BAGV RNA (genotype-2) was detected only during the 2021 outbreak in A. rufa and Emberiza calandra (1.8%, 21/1,171), although BAGV-specific antibodies (6.6%, 60/910) were detected for up to two years after the outbreak, suggesting continued exposure or low-level circulation. Overall, our findings reveal contrasting circulation dynamics of orthoflaviviruses in southern Portugal. Sustained WNV circulation implies the need to strengthen avian and human surveillance in Portugal to better assess its public health risk, while the sporadic detection of USUV and BAGV underscores the value of long-term, integrated One Health surveillance for improving early warning and preparedness for emerging mosquito-borne diseases.

Background: Hepatitis E virus (HEV) is highly endemic in several African countries with high mortality rate among pregnant women. The prevalence of HEV in Niger among pregnant women is unknown. We hereby aim to report the prevalence and risk factors of HEV among pregnant women in Niamey, Niger. Methods: Blood samples were collected from pregnant women attending antenatal consultation in three selected Health centers in Niamey. Data were collected for socio demographic characteristics using a structured questionnaire. Samples were tested using ELISA and qRT-PCR for anti- HEV IgM and RNA detection respectively. HEV associated risk factors were determined using multivariate logistic regression models. Results: A total of 491 pregnant women were enrolled with mean age of 26.5 years. The prevalence of anti-HEV IgM antibodies was 17.1%, while HEV RNA was detected in 3.46% of participants, indicating recent and active HEV infection. Anti-HEV IgM seropositivity was more frequent among women aged 26-30 years and those in the third trimester of pregnancy. Gravidity was significantly associated with HEV RNA positivity (p = 0.032). In multivariable analysis, the use of well water was independently associated with HEV infection (aOR = 75.70; 95% CI: 1.23-4643.20; p = 0.039). Conclusion: In view of the high seroprevalence of HEV among pregnant women, preventive measures including hygiene sensitization should be prioritized. Further studies should be conducted to assess the real impact of HEV infection on maternal and fetal health.

Background: Pharmacogenomic point-of-care testing (PGx POCT) enables genotype-informed prescribing at the bedside, but implementation is limited by the lack of suitable reference materials and standardised external quality assessment (EQA). Methods: Fixed lymphoblastoid cell lines were formulated as swab-based materials at defined cellular inputs for MTRNR1 POCT. Materials were evaluated using Genedrive MTRNR1 assays, quantitative PCR, acceptance quality limit batch release, workflow perturbation testing and stability studies, then deployed in an international EQA programme. Results: Materials were homogeneous, workflow-compatible and reproducible, with 100% correct genotype calls at the lowest intended manufacturing cell load. Seven EQA surveys yielded 253 participant results, with full-mark rates between 87.5 and 100%, and six test failures. Most failures occurred with low cell-input materials near the assay limit of detection. The materials supported genotype concordance, performance monitoring and identification of workflow and sample-related variability. Conclusions: Workflow-compatible cell-based reference materials combined with EQA provide a scalable framework for quality-assured PGx POCT beyond specialist laboratories. International MT-RNR1 PGx POCT EQA programme deployment demonstrated feasibility in decentralised settings. The framework may provide a model for the implementation, verification and ongoing quality assessment of PGx POCT. Further evaluation across platforms, analytes, settings and participant groups is required to establish broader applicability.

Background Metagenomic next-generation sequencing (mNGS) is increasingly adopted as a rapid and unbiased alternative to culture-based pathogen detection. However, standardized approaches for analytical validation of mNGS workflows are lacking. Validation strategies based solely on comparison with culture are inherently limited due to low diagnostic yield of culture, and the inability to derive analytical parameters such as limit of detection (LoD) from clinical samples with unknown microbial loads. Systematic analytical approaches are therefore required to define mNGS workflow performance independently of clinical comparators. In addition to diagnostics, mNGS is widely applied to microbiome profiling, where preservation of microbial diversity at low DNA inputs is a key performance criterion. Materials and Methods The analytical performance of the PaRTI-Seq mNGS workflow (Micronbrane, Taiwan), comprising host depletion, DNA extraction, library preparation, and bioinformatics analysis, was evaluated. Library-level analytical sensitivity was assessed using quantified genomic DNA (gDNA) from Enterobacter hormaechei, Candida albicans, and Mycobacterium smegmatis. Workflow-level sensitivity was evaluated using enumerated suspensions of E. hormaechei, Staphylococcus aureus, M. smegmatis, C. albicans and Aspergillus brasiliensis. Microbiome performance at low DNA input was evaluated using stool, saliva and gut mock community samples at 50ng, 5ng, and 0.5ng gDNA input and compared with an alternative low-input library preparation workflow. Results The library preparation module demonstrated analytical sensitivity down to 0.1 pg of E. hormaechei and C. albicans gDNA, and 1pg for M. smegmatis gDNA. Workflow-level LoDs were determined to be 1 CFU for E. hormaechei, 272 CFUs for S. aureus, 39 CFUs for A. brasiliensis, 36 CFUs for C. albicans and 3,702 CFUs for M. smegmatis. Differences in extraction efficiencies and extracted gDNA qualities appeared to be important determinants of workflow-level LoDs. For microbiome applications, Micronbrane Unison Ultra-low DNA input library preparation kit maintained more consistent bacterial diversity indices down to 5ng DNA input for stool and saliva samples, and the gut mock community than the comparator workflow. Conclusion This study outlines practical and generalizable analytical validation approaches for mNGS library preparation and complete workflows using defined spike-in models, providing a foundation for informed clinical interpretation and implementation of mNGS in diagnostic and microbiome applications.

Preeclampsia arises from early placental dysfunction that precedes clinical presentation, creating a need for molecular readouts that enable timely risk assessment. Placenta derived exosomes carry nucleic acid biomarkers with diagnostic potential, yet current analytical workflows rely on multistep isolation and extraction procedures that are slow and resource intensive. Here, we report Exo-LAMP, an extraction-free isothermal assay for detecting placental exosomal KISS1 mRNA, representing the first isothermal approach for exosomal mRNA detection. Placental alkaline phosphatase (PLAP) positive syncytiotrophoblast derived exosomes are selectively captured from biofluids using PLAP antibody conjugated magnetic beads within 20 minutes. Captured exosomes are treated with RNase inhibitor and lysed at 90C for 5 minutes, enabling direct RNA release while preserving transcript integrity and eliminating conventional extraction steps. The released RNA is analyzed in the same vessel by colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP), providing semi-quantitative readout within 25 to 30 minutes. The assay quantifies KISS1 mRNA across 103 to 108 exosomes per reaction, with an estimated limit of detection of about 2 exosomes and a limit of quantification of 103 exosomes. Stoichiometric analysis indicates approximately one KISS1 transcript per 27 exosomes, compared with one per 38 exosomes measured by reverse transcription quantitative polymerase chain reaction (RT-qPCR). In both the exosome-depleted serum spike-in and the plasma spike-in samples containing mixed vesicle populations, the assay detects target derived transcripts at 1% abundance and retains measurable signal at the quantification limit in the presence of up to 108 non-target exosomes. These results establish Exo-LAMP as a rapid and simplified platform for exosomal mRNA analysis that integrates selective capture, lysis, and amplification in a single workflow completed in less than 1 hour. The method provides a general framework for semi-quantitative detection of exosomal transcripts and can be adapted to other coding and non-coding RNA targets for liquid biopsy applications.

In 2024, Chicago experienced a measles virus outbreak. Air samples collected as part of routine respiratory virus surveillance from three hospitals that received measles patients were retrospectively analyzed. Measles virus was detected in air samples from one emergency department across five weeks when measles patients were present and another when no known cases were present. Two of three viral sequences derived from air samples were identical to measles patients. The source of one diverging sequence could not be identified and may represent a case not identified by traditional surveillance methods. Air sampling may supplement measles investigation, surveillance and response.

Roseoloviruses, notably human herpesviruses 6A and 6B (HHV-6A and HHV-6B), are neurotropic viruses implicated as agents in neurological disorders, including: epilepsy, multiple sclerosis, and chronic fatigue syndrome. However, the effects of roseolovirus infection on neuronal signaling and network activity are not characterized. This is, in part, due to the complexities of monitoring electrical activity in individual neurons and across neuronal connections during viral infection. This protocol describes a human induced pluripotent stem cell (iPSC)-derived neuronal culture system that employs multi-electrode array (MEA) recordings to study neurophysiological changes during viral infection. Two culture platforms are described: (a) NGN2-induced forebrain neuronal cultures; and, (b) progenitor cell-derived neuron-astrocyte mixed cultures. Cell composition in cultures is validated by immunofluorescence staining using neuronal, glial, and viral markers. Functional activity is assessed using extracellular recordings detected via the MEA2100 system. Mean firing rate, inter-spike interval, single-electrode bursting, and network burst activity are characterized between roseolovirus-infected versus uninfected (control) states. Pharmacological treatment with bicuculline, gabazine, and nicotine in conjunction with immunofluorescence is used to confirm functional responsiveness of defined neuronal neurotransmitter chemotypes. Here, we show that HHV-6A infection changes neuronal firing compared to uninfected controls. The methods/workflows described provide a strategy to study how virus infection alters neuronal excitability and may be adapted to compare the effects of different viruses on nerve cell function, infection time courses, different multiplicities of infection (MOI), and therapeutic interventions.

Consumer-grade wearables can continuously and longitudinally measure health signals such as heart rate (HR), activity, and sleep. While changes in these signals have been implicated in the early detection of infection (e.g., influenza, SARS-CoV-2) and real-time alerting systems have been developed, evidence from prospective deployment in pediatric transplant households remains limited. Here, we study the application of a resting heart rate (RHR)-based real-time alerting algorithm to detect respiratory virus infection in a cohort of solid organ transplant recipients and their household members (n=146) in a Phase IV clinical trial (NCT06161454). Reliable and efficient alerting for infection events may be particularly relevant for this population, enabling isolation, clinical management, and/or treatment. Of 31 antigen-test outcomes, nearly all (29/31, 93.5%) correspond to either influenza, COVID, or other patient-reported symptoms. We describe RHR, HR, activity, and sleep/circadian rhythm features around clinical alarm events, identifying candidate features for further investigation and algorithm improvement.

Background/Aims: In medical practice, the rising prevalence of infections in kidney transplant recipients (KTRs) is a notable concern. However, understanding the incidence and risk factors associated with infections in KTRs remains limited. This study aims to assess the incidence and factors associated infection among KTRs. Additionally, we also investigate the median time of each infection among KTRs. Methods: A retrospective cohort study was conducted from 1 January 2018- 31 December 2022 at Thailand's tertiary hospital. We performed univariable and multivariable logistic regression analyses to identify potential risk factors related infection and investigated for the incidence and median time of each infection among KTRs. Results: A total of 179 participants were included in this study. Our findings reveal the kidney from living donor was protective factor for infection among KTRs (aOR 0.063, 95% CI 0.005-0.792; p = 0.032). Urinary tract infections (UTIs)showed as the predominant pathogenic manifestation among KTRs (43%). The median time of UTIs, cytomegalovirus (CMV) infection and pneumocystis pneumonia (PCP) were 16, 57 and 94.5 days, respectively. Conclusions: Our study establishes that receiving a kidney from a living donor is a protective factor against post-kidney transplant infections. Particularly urinary tract infections were most common of infection. Monitoring urinary tract infections post-transplantation is recommended. Moreover, awareness of UTI, CMV infection and PCP should be concerned during 2-3 months after transplantation. However, due to a small sample size; we recommend to increase the number of particular participants in further investigations.

Low pathogenic avian influenza A(H7N7) virus detected in a poultry worker in Taiwan, 2026 was related to viruses circulating in Asian wild birds and possessed PB2 E627K, a mammalian adaptation marker. Enhanced surveillance for H7 viruses in poultry and rapid characterization of human cases are essential to assess pandemic potential.

GeneCapture is developing a novel, point-of-care, rapid AST technique that, when fully automated, can accomplish AST within 2 hours directly from a raw sample, representing a significant advancement in diagnostic capabilities. For this study, a panel of uropathogen isolates was collected from clinical specimens positive for urinary tract infection from various laboratories in the southern US. A selection of these isolates were then spiked into fresh clean-catch urine samples to produce mock samples covering a variety of antibiotic responses. Testing was performed on 20 strains of Escherichia coli, the cause of over 80% of UTIs, and 6 strains each of Klebsiella pneumoniae and Proteus mirabilis, the next most common causes. Each strain was tested against the susceptible, intermediate and resistant (S-I-R) breakpoints of up to 5 appropriate antibiotics including Amoxicillin-Clavulanate, Cefpodoxime, Ciprofloxacin, Nitrofurantoin and Trimethoprim-Sulfamethoxazole. The new method uses a phenotypic assessment of bacterial growth, comparing responses from bacteria treated with antibiotics at S-I-R breakpoint concentrations to untreated controls as the metric for determining susceptibility. Exposure to the drugs for only 90 minutes gave a Categorical Agreement of 98% to gold standard culture results across the resulting 247 bacteria/antibiotic pairs. Testing on separate days of nine of these strains in triplicate gave a reproducibility of 97%. The sequential steps of this novel AST - sample processing, growth, labeling, and counting - can readily be performed on an automated cartridge that is currently under development. The presented work, which took place in a laboratory setting, demonstrates the efficacy of the assay.