Ludovica Ingletto

Accurate hepatitis B virus (HBV) DNA quantification is essential for assessing viral replication and monitoring antiviral therapy. This retrospective method-comparison study evaluated the Bosphore HBV Quantification Kit against the Aptima HBV Quant Assay using 130 residual anonymized EDTA-plasma specimens, comprising 110 comparator-positive and 20 comparator-negative samples. Categorical agreement was assessed using positive, negative, and overall percent agreement (PPA, NPA, and OPA), while quantitative agreement was evaluated using a predefined ±0.5 log₁₀ IU/mL criterion, Bland–Altman analysis, Passing–Bablok regression, and Lin’s concordance correlation coefficient (CCC). PPA was 100% (110/110; 95% confidence interval [CI], 96.70–100.00%), NPA was 95.0% (19/20; 95% CI, 75.13–99.87%), and OPA was 99.23% (129/130; 95% CI, 95.79–99.98%). One confirmed qualitative discordance was observed. Overall, 109/110 paired numerical results (99.1%; 95% CI, 95.04–99.98%) were within ±0.5 log₁₀ IU/mL. Bland–Altman analysis showed a mean bias of +0.124 log₁₀ IU/mL, Passing–Bablok regression yielded Y = 0.107 + 1.019X, and Lin’s CCC was 0.991. The Bosphore assay demonstrated high categorical and quantitative agreement with the comparator across the HBV DNA concentration range evaluated, supporting its potential suitability for quantitative HBV DNA testing within the analytical workflow assessed in this study.

Background/Objectives: Modifiable dietary and lifestyle factors may contribute to gut microbiota variation but are often evaluated separately. This pilot study examined four exposures across complementary microbiome domains and used association magnitude and uncertainty to inform longitudinal study design. Methods: Twenty healthy adults (10 females and 10 males) provided a fresh-stool specimen and completed the exposure questionnaire on the day of specimen production. Exposures comprised Mediterranean diet adherence, ultra-processed food consumption frequency, physical activity, and sleep characteristics. Microbiome outcomes comprised independently recalculated OTU-level Shannon diversity (primary outcome) and Pielou’s evenness, genus-level Bray–Curtis and Aitchison distances, centered log-ratio coordinates of eight prespecified genera, and the natural-log-transformed Prevotella/Bacteroides ratio. Associations were evaluated using age- and sex-adjusted linear regression and permutational multivariate analysis of variance, with false-discovery-rate control, influence diagnostics, and robustness analyses. Scenario-based sample-size calculations were restricted to age- and sex-adjusted cross-sectional linear regression models. Results: Complete exposure and microbiome data were available for all participants. No association was supported after false-discovery-rate correction. However, the observed effect estimates were not uniformly negligible: maximum partial R² values were 0.140 across the alpha-diversity models, 0.210 for the UPF–Akkermansia association, and 0.108 for the Prevotella/Bacteroides log-ratio, whereas the maximum marginal R² for overall community composition was 0.0649. Estimates were imprecise and participant-sensitive, and robustness analyses did not alter the overall interpretation. Conservative scenarios for age- and sex-adjusted cross-sectional regression models, based on assumed partial R² values of 0.03–0.05, yielded planning benchmarks of 275–465 analyzable participants, or 306–582 enrolled participants assuming 10–20% unusable observations. Conclusions: This pilot does not establish the absence of diet–microbiota or lifestyle–microbiota relationships. Instead, it provides a reproducible framework for prioritizing hypotheses and planning adequately powered longitudinal studies using repeated, temporally aligned exposure and microbiome measurements.

Pre-analytical variables, including specimen transport media, may influence the performance of molecular assays used for respiratory pathogen detection. Universal Biomolecular Medium (UBM™) has been developed to support nucleic acid preservation and molecular testing, but evidence regarding its analytical compatibility with routinely used molecular platforms remains limited. We conducted a retrospective paired comparison of UBM™ against conventional transport and lysis media across multiple molecular platforms to evaluate the post-collection analytical compatibility of UBM. Residual respiratory specimens positive for SARS-CoV-2, influenza A virus, influenza B virus, respiratory syncytial virus, human rhinovirus, human metapneumovirus, Haemophilus influenzae, Mycoplasma pneumoniae, and Bordetella pertussis were analyzed. Paired aliquots generated from the same specimen were tested using the Allplex™ SARS-CoV-2/FluA/FluB/RSV Assay, Xpert^® Xpress CoV-2/Flu/RSV plus, Panther Fusion^® SARS-CoV-2/Flu A/B/RSV Assay, and selected Allplex™ respiratory panels. Analytical agreement and cycle threshold (Ct) values were compared using paired statistical analyses and Bland–Altman methods. UBM demonstrated complete qualitative concordance with comparator media across all evaluated targets and platforms, yielding 100% positive, negative and overall agreement. Although statistically significant Ct differences were observed for selected target–platform combinations, these shifts were generally small and did not affect qualitative result interpretation. Internal control performance remained stable across media. These findings support the post-collection analytical compatibility of UBM™ with the evaluated molecular workflows.

Acute infectious gastroenteritis (AIG) is caused by diverse bacterial and viral enteric pathogens with overlapping clinical presentations, limiting conventional pathogen-directed workflows. This retrospective before–after study evaluated the diagnostic, microbiological, operational, and economic impact of multiplex molecular syndromic testing for AIG within a regional hub-and-spoke laboratory network in Italy. Two 19-month periods were compared: a pre-implementation period based on conventional diagnostics and a post-implementation period using multiplex syndromic panels as first-line tests. Diagnostic investigations totalled 25,574 before and 24,509 after implementation. Overall test positivity increased from 5.43% to 11.59% (p < 0.001), with significant increases for bacterial and viral targets. Pathogen detection events increased from 1388 to 2839, including broader detection of Campylobacter spp., Shigella spp./enteroinvasive Escherichia coli (EIEC), Yersinia enterocolitica, Aeromonas spp., Astrovirus, Sapovirus, and Norovirus genogroups. Mean turnaround time decreased from 63 to 47 h from sample collection and from 56 to 41 h from laboratory check-in. Although direct diagnostic costs increased moderately, total laboratory costs decreased when personnel costs were included, and cost per diagnostic detection declined from €233.80 to €120.86. Multiplex syndromic testing improved microbiological detection, diagnostic efficiency, turnaround time, and laboratory organisation in routine AIG diagnosis.

Sexually transmitted infections (STIs) remain a major global public health concern, with more than one million new cases acquired daily and increasing antimicrobial resistance compromising effective control strategies. Rapid, accurate and multiplex molecular diagnostics are therefore essential to support timely clinical management and public health surveillance. This study evaluated the analytical and clinical performance of the cartridge-based STANDARD™ M10 STI Panel (SD Biosensor, Republic of Korea), a fully automated, random-access real-time PCR assay capable of detecting eight STI-related pathogens within approximately 64 min. A total of 150 residual clinical specimens were retrospectively analysed, including vaginal, rectal, urethral and oropharyngeal swabs, as well as seminal fluids, and results were compared with those obtained using the Allplex™ STI Essential Assay (Seegene, Republic of Korea), which shares six common targets. The STANDARD™ M10 STI Panel demonstrated high diagnostic accuracy, with sensitivity ranging from 89.5% to 100%, specificity from 98.2% to 100% and overall accuracy between 98% and 100%. Agreement between the two assays was almost perfect, with Cohen’s κ values ranging from 0.91 to 1.00. Analytical sensitivity was further confirmed through verification of the limits of detection using quantified reference standards. Although validated for urine samples, the assay also showed robust performance on alternative clinical matrices, particularly vaginal swabs. Overall, these findings indicate that the STANDARD™ M10 STI Panel represents a reliable and practical tool for STI diagnosis, combining rapid turnaround time, minimal hands-on requirements and broad pathogen coverage in both centralized and near-patient testing settings.

Transboundary animal diseases (TADs), including zoonoses, can be introduced in non‐endemic areas through animal trade and uncontrolled movements during times of conflict and migration. Sheep and goat pox (SGPX) and peste des petits ruminants (PPRs) were, respectively, reported in Europe in 2018 and 2010. EFSA emphasised the risk of spread into Europe through informal trade and unmonitored migration routes that may involve the informal transport of small ruminants. Using a One Health conceptual framework, which considers the interconnectedness of human, animal, and environmental health, this study analysed the epidemic trends of the two selected diseases during 2014–2025, with particular focus on the role of human migrations in influencing their spread along major migratory routes in the Balkan Peninsula. The conceptualisation of animals in migration settings was investigated through the scrutiny of policies and semi‐structured interviews to experts and people who travelled the route. The presence of small ruminants, companion and pest animals was confirmed in formal and informal camps. The results underscore the relevance of animals in the migration contexts of the Balkan routes, as well as the risks associated with their presence in settlements in the absence of an adequate management. Our findings seem to suggest that human migration along the Balkan routes is not directly responsible for the spread of TADs, as there is no evidence of significant animal movements accompanying migrants. However, our results show a possible role of informal trade networks following human migration paths along the Balkan route, while a possible role of informal trade networks, following human migration paths along the Balkan route, deserves to be further investigated.

Strong epidemiological evidence suggests that onchocerciasis may be associated with epilepsy—hence the name onchocerciasis-associated epilepsy (OAE). However, the pathogenesis of OAE still needs to be elucidated, as recent studies have failed to detect Onchocerca volvulus in the central nervous system of persons with OAE. Therefore, it was suggested that a potentially neurotropic virus transmitted by blackflies could play a role in triggering OAE. To investigate this hypothesis, adult blackflies were collected in an onchocerciasis-endemic area with a high OAE prevalence in the Ntui Health District, Cameroon. A viral particle-based shotgun sequencing approach was used to detect viral sequences in 55 pools of 10 blackflies. A very high abundance of viral reads was detected across multiple (novel) viral families, including viral families associated with human disease. Although no genomes closely related to known neurotropic viruses were found in the blackfly virome, the plethora of novel viruses representing novel species, genera and even families warrant further exploration for their potential to infect vertebrates. These results could serve as a first step for studying the viruses associated with the haematophagous blackfly, which also could be present in their nematode host O. volvulus. Exploring the diversity of viruses in blackflies should be included in the active surveillance of zoonotic diseases.