Journal of Clinical Virology Plus

Introduction: Herpes simplex virus type 1 (HSV-1) is highly prevalent worldwide, making accurate serological testing essential for both clinical diagnosis and epidemiological surveillance. Automated chemiluminescent immunoassays (CLIAs) offer operational advantages over enzyme-linked immunosorbent assays (ELISAs); however, their diagnostic performance relative to Western blot (WB) confirmation in high-prevalence settings remains insufficiently characterized. Hypothesis/Gap Statement: The comparative diagnostic accuracy of CLIA- and ELISA-based assays for HSV-1 IgG detection, when benchmarked against a WB reference standard in endemic populations, remains unclear. Aim: This study aimed to evaluate HSV-1 IgG seroprevalence and diagnostic performance of one CLIA and two ELISA platforms using Western blot as the reference method. Methodology: Four hundred archived serum samples from adult male craft and manual workers in Qatar were tested using the Mindray CL-900i CLIA, HerpeSelect ELISA, NovaLisa ELISA, and Euroimmun Western blot. Seroprevalence, diagnostic accuracy, and interassay agreement were assessed using WB as the reference standard, with equivocal and indeterminate results excluded from analysis. Results: HSV-1 IgG seroprevalence estimates were comparable across assays: HerpeSelect 72.5%, Mindray 70.5%, NovaLisa 66.3%, and Western blot 66.5%, with no statistically significant differences (all p > 0.05). The Mindray CLIA demonstrated the highest diagnostic performance (sensitivity 95.7%, specificity 88.9%, accuracy 93.4%) and strong agreement with Western blot ({kappa} = 0.85). HerpeSelect showed substantial agreement ({kappa} = 0.81), while NovaLisa exhibited lower specificity. Conclusion: CLIA- and ELISA-based assays produced comparable HSV-1 seroprevalence estimates in this high-prevalence population; however, diagnostic accuracy varied across platforms. The CLIA platform demonstrated the strongest agreement with Western blot, supporting its use in high-throughput settings, while confirmatory testing remains important to minimize misclassification.

BackgroundImmunocompromised (IC) individuals are at increased risk for persistent SARS-CoV-2 infections and can develop new viral mutations and lineages not seen in the community. In this case report, a persistent SARS-CoV-2 infection (330 days) in an IC patient is examined for viral mutations and mutations associated with cryptic lineages. Case PresentationThe patient was followed in a longitudinal study examining persistent SARS-CoV-2 in IC patients. The patient provided stool and nasal swab samples biweekly until 28 days post-enrollment, then monthly, and then quarterly after 12 month post enrollment until the participant was no longer positive for SARS-CoV-2. Staff performed RT-qPCR and viral sequencing on the samples. Viral mutations from the XBK lineage were already present in the initial sample. By the end of the infection period, there were 40 fixed consensus changes from XBK of which two mutations were typical for cryptic lineages. Mutations increased steadily over time, with most mutations fixed by day 253, including the cryptic typical mutations. ConclusionThis case demonstrates the potential for persistent SARS-CoV-2 infections to develop mutations and lineages in IC patients and highlights the need for continued SARS-CoV-2 monitoring and treatment in this vulnerable population.

BackgroundHepatocellular carcinoma (HCC) is a leading cause of cancer-related death in sub-Saharan Africa (SSA). Differentiating primary HCC from metastatic liver tumors remains a significant diagnostic challenge. Understanding the prevalence and clinical predictors of HCC is crucial for improving diagnosis and patient care. This study examined the prevalence of hepatitis B virus (HBV), hepatitis C virus (HCV), and HCC, and clinical predictors of HCC. MethodsWe used immunohistochemical markers on archived liver tumor biopsies and analyzed the data using descriptive and logistic regression analysis. ResultsAmong 58 liver carcinoma cases, 37.9% had HCC, and 62% had metastatic liver carcinoma (MLC). HCC was most common (61.5%) among middle-aged adults (50-59 years). HCC was more frequent in males (47.2%) than in females (22.7%). Over half of the patients (51.7%) tested positive for HBV. HCC was more prevalent in HBV-positive patients than HBV-negative ones (43.3% vs 32.1%). Hepatic fibrosis was identified in 27.6% of cases. HCC was more common in patients with fibrosis (56.2%) than in those without (31%). HCV infection was rare (6.9%) in this study. In multivariable logistic regression analysis, none of the examined predictors reached statistical significance (P>0.05). Patients aged 50-59 years, males, those with HBV infection, and hepatic fibrosis showed higher odds of HCC. Hepatocyte Paraffin-1 (Hep Par-1) demonstrated 97% specificity and a 95% positive predictive value (PPV) for differentiating HCC from MLC. The combined marker pattern of Hep Par-1 positive and AE1/AE3 negative was highly predictive of HCC (100% specificity, 100% PPV, and 93.2% diagnostic accuracy). ConclusionsOur findings indicate that while the assessed risk factors tend to show directional association with HCC, as expected, larger studies are needed to determine their independent effects. The combined Hep Par-1 AE1/AE3 immunophenotype is more accurate than either marker alone. Therefore, this combined test is a valuable diagnostic tool for confirming HCC in resource-limited settings.

ObjectiveThis study investigates the prevalence of human pegivirus (HPgV) in SARS-CoV-2-positive patients within the context of viral co-infections that may modulate COVID-19 outcomes and assesses whether HPgV co-infection is associated with COVID-19 severity. HPgV is a widely circulating but rarely monitored human virus with documented immunomodulatory effects in other viral infections, including HIV and Ebola. While HPgV prevalence is low in the general U.S. population (1-2%), it rises markedly in the setting of chronic viral co-infections, particularly HIV (15-40%). Given its immunologic effects and persistence, HPgV represents a biologically plausible but unexplored viral co-factor SARS-CoV-2 infection. MethodsWe analyzed four cohorts: SARS-CoV-2-positive individuals, ICU patients with respiratory symptoms but SARS-CoV-2-negative, HIV-positive individuals as a positive control for HPgV detection, and uninfected controls. ResultsHPgV prevalence in COVID-19 patients was low (2.1%) and comparable to population estimates. As expected, HPgV prevalence was substantially higher in the HIV cohort (34%), validating assay performance and cohort stratification. Among HPgV-positive COVID-19 cases, most experienced mild disease, with directional trends toward reduced severity despite high baseline risk factors. Healthcare workers in the control group showed unexpectedly elevated HPgV prevalence (9.6%). ConclusionsHPgV is an unmonitored but widely circulating viral co-infection in humans that may influence host responses to SARS-CoV-2. Although limited by small numbers, our findings support further investigation of HPgV and other immunomodulatory viral co-infections in COVID-19. This study suggests that HPgV co-infection may influence COVID-19 outcomes, warranting further investigation. HighlightsO_LISystematic screening of human pegivirus (HPgV), an unmonitored viral co-infection, in COVID-19 (n = 634). C_LIO_LIHPgV prevalence in COVID-19 mirrored population estimates but was markedly enriched in HIV (positive control). C_LIO_LIHPgV-positive COVID-19 cases showed trends toward milder clinical outcomes. C_LIO_LIFindings highlight the potential relevance of immunomodulatory viral co-infections in SARS-CoV-2 infection. C_LI Executive SummaryThis study evaluated the prevalence of human pegivirus (HPgV) co-infection among SARS-CoV-2-positive patients as part of a broader effort to understand how concurrent viral infections may influence COVID-19 severity. HPgV is a largely unmonitored, persistent human virus with well-described immunomodulatory effects in other viral infections, yet it has not been systematically evaluated in COVID-19. We therefore screened HPgV prevalence across COVID-19 cases, comparator cohorts, and an HIV-positive cohort as a positive control due to the well-established high prevalence of HPgV in HIV infection. Our findings indicate that HPgV prevalence in SARS-CoV-2-positive and -negative hospitalized individuals are consistent with the general U.S. population range (approximately 1-5%). Healthcare professionals exhibited a higher HPgV prevalence ([~]10%), suggesting that repeated occupational viral exposures may influence infection rates. While limited by small numbers, HPgV co-infection in COVID-19 cases was associated with directional trends toward reduced disease severity, warranting further longitudinal and mechanistic investigation. Editors summaryThis study identifies human pegivirus as a widely circulating, unmonitored viral co-infection in COVID-19 with potential relevance to disease severity.

BackgroundIn 2004, South Africas public health system faced the dual challenge of rapidly scaling up antiretroviral therapy (ART) while reducing the cost of laboratory monitoring. At the time, conventional CD4 testing methods were expensive, labour-intensive, and impractical for sustaining a national testing network. This study aimed to assess the financial impact and cost savings associated with the implementation of the PanLeucogated CD4 (PLG/CD4) enumeration method between 2004 and 2024 in the public-sector in South Africa. MethodsA longitudinal cost analysis was conducted using annual test volumes and state tariffs for PLG/CD4 testing and the 4-colour CD3/CD4/CD8/CD45 T-cell enumeration reference method. Annual cost savings were calculated in United States Dollars (USD) by applying historical South African Rands (ZAR) to United States Dollars (USD) exchange rates. The state prices for tariff codes PLG/CD4 and the reference method were provided by calendar year in ZAR and converted to USD based on the prevailing exchange rate. The USD test prices were multiplied by annual test volumes. Cost savings were calculated by multiplying annual test volumes and the difference in test prices in USD (difference between PLG/CD4 and the reference method). ResultsThere were 50,745,848 PLG/CD4 tests performed over 20-years. The cost-per-test of PLG/CD4 was consistently lower than the reference method, ranging from $4,06 to $9,40, compared to $13,06 to $28,21. Cumulative national savings amounted to USD 626 million. The peak annual savings of $64,6 million occurred in 2011, coinciding with the height of ART enrolment. Cost savings persisted despite a doubling in the exchange rate over the study period. ConclusionThe PLG/CD4 implementation enabled cost-efficient, scalable, quality-assured CD4 testing as part of the national HIV response, reducing reliance on complex/costly technologies while improving coverage. These findings support the critical role of context-specific diagnostic innovation to strengthen health system resilience.

BackgroundNeutralising antibody titres are widely used as key immunogenicity endpoints in Ebola virus (EBOV) vaccine and monoclonal antibody clinical trials. However, direct comparison of results across studies remains challenging due to the use of heterogeneous neutralisation platforms, ranging from pseudotyped viruses to live EBOV assays. These limitations restrict assay standardisation, validation, scalability, and compliance with good clinical laboratory practice (GCLP), particularly in outbreak-prone and resource-limited settings. There is an unmet need for neutralisation assays that combine biological authenticity with clinical-trial compatibility. MethodsWe developed and optimised a fluorescence-based microneutralisation assay using a biologically contained EBOV lacking the essential VP30 gene (EBOV{Delta}VP30), enabling multi-cycle viral replication under containment level 2 conditions. Using a defined panel of serum samples from Ebola virus disease survivors and EBOV-negative controls, we benchmarked EBOV{Delta}VP30 neutralisation titres against previously generated data obtained with wild-type EBOV and pseudotyped virus platforms. Assay performance was evaluated in terms of sensitivity, reproducibility, discrimination between positive and negative samples, and correlation with live virus neutralisation. Calibration was performed using the WHO International Standard for anti-EBOV immunoglobulin. FindingsThe EBOV{Delta}VP30 microneutralisation assay robustly distinguished EBOV survivor sera from negative controls (p < 0{middle dot}0001) and demonstrated a strong correlation with live EBOV neutralisation titres (Spearman {rho} = 0{middle dot}8725). This correlation exceeded that observed for HIV-1-based pseudotyped assays and for the vesicular stomatitis virus-based platforms. The fluorescence-based read-out showed comparable sensitivity to conventional immunostaining, supporting its suitability for high-throughput and standardised implementation. Importantly, assay conditions were compatible with BSL-2 laboratories and GCLP-aligned workflows. InterpretationBiologically contained EBOV{Delta}VP30 provides a clinically relevant and scalable alternative to existing neutralisation platforms, bridging the gap between pseudotyped assays and wild-type virus testing. By improving biological relevance while maintaining accessibility and standardisation, this assay has the potential to enhance comparability of immunogenicity data across EBOV vaccine and therapeutic antibody (pre-)clinical trials, aligning with global outbreak preparedness and trial harmonisation objectives. FundingStated in acknowledgement section of manuscript. Research in contextO_ST_ABSEvidence before the studyC_ST_ABSBefore starting this study, we reviewed published work on how neutralising antibodies against Ebola virus are measured in vaccine and monoclonal antibody research. We searched PubMed, Web of Science, and reference lists of key review papers for studies published up to mid-2025, without restricting by language. Search terms included "Ebola virus", "neutralising antibodies", "neutralisation assay", "pseudovirus", "live virus", and "clinical trials". We focused on studies describing neutralisation tests using wild-type Ebola virus as well as commonly used pseudotyped virus systems. From this body of evidence, neutralisation assays using wild-type Ebola virus are considered the most biologically relevant but can only be performed in biosafety level 4 laboratories. This limits their availability, scalability, and use in clinical trials. Pseudotyped virus assays can be performed under lower biosafety conditions and are widely used, but multiple studies have reported variable performance and inconsistent agreement with live virus results. Although biologically contained Ebola viruses have been developed and used in laboratory research, their application as neutralisation assays and their direct comparison with both live virus and pseudotyped systems using the same human serum samples had not been systematically studied. As a result, it remained unclear whether such systems could support reliable immunogenicity assessment in clinical trials. Added value of this studyThis study shows that a biologically contained Ebola virus lacking the VP30 gene can be used to measure neutralising antibodies in a robust and scalable way under biosafety level 2 conditions. By directly comparing this system with wild-type Ebola virus and widely used pseudotyped assays using the same set of human serum samples, we demonstrate that neutralisation results obtained with the biologically contained virus closely align with those of the wild-type virus reference assay. The assay reliably distinguishes samples from Ebola survivors and uninfected individuals and can be read using different detection methods, making it compatible with GCLP-aligned workflows and suitable for further qualification and validation in support of clinical development. This work provides clear evidence that biologically contained Ebola virus can combine biological relevance with practical usability. Implications of all the available evidenceTogether with existing evidence, our findings indicate that biologically contained Ebola virus offers a valuable new option for measuring neutralising antibodies in vaccine and monoclonal antibody clinical trials. By reducing reliance on high-containment laboratories while preserving key features of authentic virus infection, this approach can improve the consistency and comparability of immunogenicity data across studies and sites. Broader use of such assays could support better decision-making during clinical development and strengthen outbreak preparedness. More generally, this work highlights how biologically contained viruses can help advance research licensure of medical countermeasures for high-consequence pathogens in ways that are directly relevant to human health.

BackgroundDengue virus infection remains a significant public health concern in India, with changing serotype dynamics influencing disease epidemiology. Understanding local serotype distribution and clinical characteristics is crucial for effective disease management and surveillance. ObjectivesTo determine the prevalence of dengue virus serotypes and analyze their clinical characteristics among NS1-positive patients at a tertiary-care hospital in Gujarat, India. MethodsA cross-sectional study was conducted on NS1-positive dengue patients admitted to AIIMS Rajkot from September 2023 to November 2024. Real-time reverse transcription polymerase chain reaction (RT-PCR) was performed for serotype identification. Clinical and demographic data were collected and analyzed. ResultsNS1-positive patients (70) were confirmed by RT-PCR. DENV-2 was the predominant serotype (53 cases, 75.7%), followed by DENV-1 and DENV-3 (7 cases each, 10.0%), and DENV-4 (2 cases, 2.9%). One co-infection case (DENV-2 + DENV-3) (1.4%) was identified. The mean age was 27.7 {+/-} 14.4 years, with male predominance (58.6%). Young adults (19-35 years) were most affected (45.7%), followed by pediatric patients [&le;]18 years (32.9%). Severe dengue occurred in only one case (1.4%), while hospitalization was required in 25 cases (35.7%). All patients presented with fever, chills, headache (50%), rashes (56%), and malaise (56%), being the most common associated symptoms. ConclusionsDENV-2 showed clear predominance in the Rajkot region during the study period, with low rates of severe disease. The significant pediatric and young adult involvement highlights the need for targeted prevention strategies. These findings contribute to the understanding of regional dengue epidemiology and support evidence-based surveillance and control measures.

Background: Although the relaxation of COVID-19 containment measures in China has altered the transmission dynamics of respiratory pathogens, regional data on post-pandemic epidemiological characteristics remain limited.Objective: This study aimed to investigate the pathogen spectrum and epidemiological characteristics of acute respiratory infections (ARIs) in Quzhou City from 2023 to 2024, providing a scientific basis for local prevention and control strategies.Methods: A total of 2,800 respiratory specimens were collected from November 2023 to July 2024, comprising 1,960 influenza-like illness (ILI) cases from outpatient/emergency departments and 840 severe acute respiratory infection (SARI) cases from inpatient departments. All samples were tested for 13 common respiratory pathogens using multiplex fluorescence quantitative PCR. Etiological and epidemiological analyses were performed based on detection results and case information. Results: The overall ARI positivity rate was 59.28% (1,660/2,800), with a male-to-female ratio of 1.07:1 (1,447/1,353). The three most prevalent pathogens were influenza virus (Flu, 23.21%, 650/2,800), Streptococcus pneumoniae (SP, 13.14%, 368/2,800), and adenovirus (ADV, 8.39%, 235/2,800). Single pathogen infections accounted for 73.55% (1,221/1,660) of positive cases, while co-infections with two or more pathogens accounted for 26.45% (439/1,660), yielding an overall co-infection rate of 15.68% (439/2,800). No significant gender difference was observed in detection rates. However, significant differences were found across case types, temporal periods, age groups, and geographic regions (P < 0.01). Children aged [&le;]5 years exhibited the highest positivity rate (78.00%, 378/525), while adults aged [&ge;]65 years showed the lowest (34.53%, 144/417). Among surveillance regions, Kaihua County had the highest positivity rate (72.47%), and Changshan County the lowest (40.55%). Conclusions: Multiple respiratory pathogens and co-infections are prevalent in Quzhou City, with distinct age-specific and seasonal patterns. These findings underscore the need for continuous multi-pathogen surveillance and integrated prevention strategies for influenza and other respiratory infectious diseases in the post-pandemic era.

Molecular and microscopy-based diagnostic capacity is often insufficient or unavailable in places where infectious disease burdens are highest, such as in West Africa. Rapid diagnostic testing (RDT) can provide quick and affordable diagnoses of common infections but is an imperfect solution due to limitations around detecting and dealing with false negative and false positive results. An alternative to RDT is unbiased metagenomic sequencing for pathogen surveillance. Here, we present data from unbiased metagenomic sequencing used to identify causes of undiagnosed febrile illness in Jos, Plateau State, Nigeria. Proof of concept for this approach has been demonstrated by several groups who have identified epidemic and endemic viral diseases like Lassa fever, yellow fever, and Chikungunya. Here, we show that unbiased deep sequencing and metagenomic analysis can be used to identify RNA viruses in clinical samples. We sequenced RNA from sera of patients (n = 343), many of whom were acutely febrile (76 %) in a survey of clinics in Jos. We detected five human-infecting viruses in 39 (11 %) specimens. Among these were hepatitis B virus, human pegivirus, and several anelloviruses. While most of the viruses identified are unlikely to cause clinical symptoms in the patients we sampled, their presence demonstrates the validity of our approach. Additionally, our sequencing data allowed us to identify genetic material from potentially pathogenic bacteria, another possible etiological agent of febrile illness.

Background: Although the hemagglutination inhibition (HAI) titer remains the gold standard correlate of protection against influenza, it does not fully capture the broader antibody responses that contribute to immunity. Methods: We analyzed immune responses in paired pre-infection and convalescent sera from 306 RT-PCR-confirmed A/H3N2 infections from two household studies (2014-18) in Managua, Nicaragua. Antibody responses were measured by HAI and enzyme-linked immunosorbent assays (ELISAs) against full-length hemagglutinin (HA), the HA stalk, and neuraminidase (NA). Participants were classified as HAI responders ([&ge;]4-fold HAI rise), alternate responders (no HAI rise but [&ge;]4-fold boost in [&ge;]1 ELISA), or no-response individuals (no [&ge;]4-fold rise in any assay). We compared demographic, clinical, and pre-infection antibody characteristics across these groups. We also analyzed predictors of an NA response. Results: Overall, 77% of participants had HAI seroconversion or a 4-fold rise. Among the 23% HAI non-responders, 62% had alternate antibody responses. No-response individuals had the highest pre-infection HAI and full-length HA titers (p < 0.0001), the lowest viral loads, and the fewest fever or influenza like illness (ILI) symptoms (p < 0.01). An NA response was more common among symptomatic individuals (p = 0.0483) and those with low or high baseline NA titers. Conclusions: High baseline HAI titers can limit detectable 4-fold rises and are associated with milder illness. Evaluating additional immune responses may capture a more complete picture of the host response to infection, thereby improving surveillance and informing vaccine development. Keywords: Influenza A/H3N2; Hemagglutination inhibition (HAI); Neuraminidase antibodies; symptomatic vs asymptomatic infection; correlates of protection.

Objective To evaluate physician knowledge, attitudes, and practices regarding viral exanthem diagnosis and mandatory reporting requirements among practicing physicians in major metropolitan regions of Florida. Study Design An IRB-exempt cross-sectional survey was distributed via REDCap to licensed physicians and residents in family medicine, internal medicine, pediatrics, and infectious disease across Florida. The 19-question survey assessed demographic characteristics, knowledge of viral exanthem diagnosis (measles, rubella, roseola), reporting requirements, physician attitudes, and clinical practices. Knowledge scores were compared by specialty using ANOVA with Tukey post-hoc analysis. Multivariate analysis and linear regression assessed associations between physician confidence and knowledge scores. Results A total of 162 physicians responded, with 146 complete responses included in analysis. Participants included pediatrics (n=74), family medicine (n=48), and internal medicine (n=24). The overall mean knowledge score was 78.5% (SD 20.5). Pediatricians demonstrated the highest scores (82.7%) compared with internal medicine (76.4%) and family medicine (73.3%), with pediatricians scoring significantly higher than family physicians (p=0.04). Differences in vignette-based diagnostic knowledge and mandatory reporting knowledge were not statistically significant across specialties. Roseola was the most commonly diagnosed viral exanthem (66%), followed by measles (30%) and rubella (17%). Most physicians (91.4%) expressed interest in additional training. Conclusions Although overall physician knowledge of viral exanthem diagnosis and reporting was high, clinically meaningful gaps remain, particularly in differentiating similar rash presentations. Pediatricians demonstrated higher knowledge scores than family physicians. Enhanced physician education may improve diagnostic accuracy and public health reporting as vaccination rates decline and outbreaks of vaccine-preventable viral exanthems increase.

Cetacean pathology is a cornerstone for population and marine ecosystem health monitoring, allowing clear differentiation among natural and anthropogenic threats. Previous studies in the Canary Islands reported natural causes of death in 59.4% (1999-2005) and 81% (2006-2012) of stranded cetaceans, versus anthropogenic causes in 33.3% and 19%, respectively. This study aimed to determine the causes of death (CD), pathologic findings, and epidemiological patterns of 316 cetaceans stranded in the Canary Islands between 2013 and 2018. The CDs were classified in pathologic entities (PEs) emphasizing natural versus anthropic origins. Of 316 animals, 224 (70.9%) from 18 species were suitable for pathological investigations. Among natural PEE, natural pathology associated with good nutritional status (NP-GNS) and natural pathology associated with significant loss of nutritional status (NP-LNS) represented 43/224 (19.2%) and 36/224 (16%) cases, respectively. Natural pathology with undetermined nutritional status (NP-UNS) occurred in 19/224 (8.5%) animals. Intra- and interspecific traumatic interactions (ITI) represented 30/224 (13.4%) cases, followed by neonatal/perinatal pathology (NPN) 19/224 (8.5%) and live-stranding stress and/or capture myopathy (LS-CM) 18/224 (8%). Infectious and parasitic diseases predominated in natural PEs. Anthropogenic PEs included interaction with fishing activities (IFA) in 17/224 (7.6%) cases, vessel collisions (VC) in 9/22 (4%) cases, and foreign body-associated pathology (FBAP) in 3/224 (1.3%) animals. Overall, anthropogenic causes accounted for 12.9% of deaths, natural causes for 73.6%, and the CD could not be established in 30/194 (13.4%) cases. This study reaffirms the trends concerning recognized PEs (NP-GNS, NP-LNS, NP-UNS, ITI, NPN, LS-CM, IFA, VC, and FBAP), expands the body of knowledge on cetacean pathology in the Canary Islands, and reports novel findings including mixed infections, clostridiosis in uncommon species, uremic syndrome secondary to urethral nematodiasis, gas embolism in unusual species, epibiont stomatitis, congenital musculo-skeletal malformations, or neoplastic processes. These findings advance understanding of cetacean mortality patterns and support conservation and management strategies.

BackgroundMpox has spread to 35 countries in Africa, yet many face challenges achieving nationwide PCR-based testing due to cost and limited access in remote and rural areas. Point-of-care antigen-based rapid diagnostic tests (AgRDTs) may help improve diagnostic access. ObjectivesTo evaluate the diagnostic accuracy of five mpox AgRDTs manufactured by Beijing Hotgen Biotech (China), Contipharma (Belgium), Hangzhou Testsea Biotechnology (China), Guangdong Wesail Biotech (China), and NG Biotech (France). MethodsDiagnostic accuracy was assessed using 190 lesion swabs from suspected mpox cases in the Democratic Republic of the Congo, with results compared against the RADI FAST Mpox PCR assay (KH Medical, South Korea). Analytical sensitivity was evaluated using a clade Ib MPXV isolate from the WHO Biohub held by the Geneva University Hospitals, Switzerland. ResultsThe best-performing assay, the Monkeypox Virus Ag Test Kit (Guangdong Wesail Biotech), demonstrated a sensitivity of 77.3% (95% CI: 68.0-84.5) and specificity of 93.5% (95% CI: 86.6-97.0). The Hangzhou Testsea assay showed comparable performance (sensitivity 72.2%, specificity 93.5%). Beijing Hotgen and Contipharma assays exhibited moderate sensitivity (59.8% and 50.5%, respectively) with high specificity (96.8% and 95.7% respectively), while the NG Biotech assay showed the lowest sensitivity (39.2%) but similarly high specificity (96.8%). Analytical testing revealed no major differences across assays, though Guangdong Wesail demonstrated the highest analytical sensitivity, detecting clade Ib virus at a Ct of 28.3. ConclusionSeveral AgRDTs show high positive predictive values for mpox screening in high-prevalence settings, where positive test results may support confirmation but negative results cannot rule out infection. Further multicentre prospective studies are needed to define appropriate use cases as countries transition to sustainable mpox control. Research in contextO_ST_ABSEvidence before this studyC_ST_ABSAccess to decentralized mpox diagnostics remains limited in many African countries due to the cost, infrastructure, and turnaround time associated with PCR, particularly in remote settings. Antigen-based rapid diagnostic tests (AgRDTs) could expand access to point-of-care testing, but data supporting their clinical performance has been limited. We searched PubMed and medRxiv on Dec 31, 2025, using the following search string: (mpox OR monkeypox) AND (diagnostic* OR point of care OR lateral flow assay OR rapid antigen test OR AgRDT). Early evaluations of mpox AgRDTs reported very low sensitivity. A large prospective, multicentre field study in Uganda and the Democratic Republic of the Congo (DRC) evaluating a single mpox AgRDT for research use only using lesion swabs reported an overall sensitivity of 70.4%, with substantial variation by country (81.9% in Uganda vs 55.1% in DR Congo), age, and viral load, and specificity of 89.3%. Independent comparative evaluations of multiple commercially available AgRDTs with regulatory approval--particularly in clade I-endemic African settings--have been lacking. Added value of this studyThis study provides the first head-to-head clinical and analytical evaluation of five commercially available mpox AgRDTs using skin or mucosal lesion swab specimens from suspected cases in DRC. By assessing all assays against a molecular reference test under identical conditions and incorporating analytical testing with a clade Ib reference virus isolate, we demonstrate substantial variability in performance between manufacturers tests kits. Two assays achieved sensitivity against PCR of above 70% with moderate specificity (93.5%), comparable to or exceeding results reported in prior field studies, while others showed moderate to poor sensitivity. Analytical testing identified meaningful differences in detection limits, with the best-performing assay detecting clade Ib virus at viral loads corresponding to a Ct value of 28.3. These findings provide critical data to inform further field-based validation projects, assay selection and procurement decisions for use of such AgRDTs in endemic settings. Implications of all the available evidenceAcross studies, mpox AgRDTs consistently demonstrate high specificity but variable and often insufficient sensitivity to function as standalone diagnostic tools. Prior studies have provided evidence that sensitivity is highest for samples with Ct values 25 or lower, and the present comparative evaluation show that positive AgRDT results may be able to reliably confirm mpox infection in high-prevalence settings, whereas negative results cannot safely exclude disease. The heterogeneity in performance between assays underscores the need for field-based independent evaluations of most promising tests before their large-scale deployment, and cautions against treating AgRDTs as a uniform diagnostic test category. As countries transition from emergency response to sustainable mpox control, AgRDTs may have a complementary role in decentralized screening, outbreak confirmation, and triage, provided confirmatory PCR remains available. Further multi-centre prospective studies and continued assay optimization are essential to ensure antigen-based diagnostics can be used to meaningfully strengthen mpox surveillance, preparedness and outbreak response.

IntroductionDengue has been prevalent in a regular fashion in Bangladesh and Chattogram for the last 6-7 years and is showing some serotype twisting. So, the objectives of the present study were to explore the burden of dengue serotypes in Chattogram. MethodsIn this study, 223 Dengue RT-PCR positive patients were evaluated for serotyping. Gender and age group, along with cycle threshold (CT) values, were also collected. Data after collection were compiled, analyzed, and plotted in Microsoft Excel and GraphPad Prism 10.4. Ethical clearance was taken to conduct the study. ResultsAmong 223 patients analyzed, males and females were found near equal (113 and 110). Middle-aged patients were more than the extremes of age. The mean {+/-} SD of age was 33.55 {+/-} 13.67 years. Regarding serotype distributions, isolated Den 1, Den 2 and Den 3 were found 1.3%, 73.1% and 6.7%, respectively. Concurrent infections with multiple serotypes were observed in several patients, most notably the Den 2 and Den 3 combination, which accounted for 14.3% (n=32) of the cases. Other co-infections were less frequent: the Den 1 and Den 2 pairing appeared in 3.6% (n=8) of the cohort, while triple-serotype infections (Den 1, 2, and 3) and Den 3/Den 4 pairings were rare, each occurring in only 0.4% of patients. Statistical analysis of CT values revealed no significant sex-based differences for Den 2 and Den 3. However, significant variations in CT values were observed when comparing Den 1 against both Den 2 and Den 3 (p < 0.05). In contrast, the difference between Den 2 and Den 3 Ct values remained statistically insignificant. ConclusionIn the year 2025, Dengue serotypes 2 and 3 were found to be the most prevalent, both in isolated or in combinations and Den 1 and Den 4 were found minimum. Exposure to multiple serotypes and twisting from one serotype to another might influence the dengue outcome in future, which needs further exploration.

BackgroundRespiratory tract infections range from asymptomatic colonisation to an invasive disease. Recent studies suggest that nasopharyngeal microbiota may influence this variability. Emerging evidence points to Dolosigranulum pigrum, a nasopharyngeal commensal, as a potentially protective bacterium. This study aimed to identify variables associated with the presence of D. pigrum in the nasopharynx of children with varying respiratory health statuses. MethodsNasopharyngeal aspirates were collected from children <18 years who were asymptomatic (n=65), had banal viral infection (n=48), or Invasive Pneumococcal Disease (IPD) (n=27). The presence of D. pigrum was defined as >0.1% of total sequences obtained by 16S rRNA gene sequencing. Variables included sex, breastfeeding, delivery mode, S. pneumoniae carriage, respiratory viruses and clinical features. ResultsAmong 140 children (73 males, 67 females), D. pigrum was detected in 79 (56.4%): 44/65 in the healthy group; 26/48 of viral and 9/27 IPD cases. Multivariate analysis revealed significant associations with health status and sex. Healthy children were more likely to carry D. pigrum than IPD cases (44/79 vs. 26/79; p= 0.028). Males were more frequently D. pigrum carriers than females (48/79 vs. 31/79; p= 0.033). ConclusionD. Pigrum was associated with respiratory health, being more prevalent in healthy children, and showed potential sex-related differences.

BackgroundRespiratory syncytial virus (RSV) is a major cause of lower respiratory tract infections in children, often leading to hospitalisation in infants. In low-resource settings where routine RSV diagnostics are unavailable, clinical overlap with bacterial pneumonia frequently results in unnecessary antibiotic use, contributing to antimicrobial resistance. ObjectiveTo evaluate the frequency and clinical determinants of antibiotic use among RSV-positive children under two years at a tertiary hospital in Ghana. MethodsThis cross-sectional study was conducted from June to November 2023 at the Department of Child Health, Korle Bu Teaching Hospital. Children with acute respiratory illness were enrolled and tested for RSV using molecular point-of-care and reverse transcriptase-polymerase chain reaction methods. Antibiotic use and clinical characteristics were analysed among RSV-positive cases. ResultsOf 128 children enrolled, 72 (56.2%) tested positive for RSV. Among these, 48 (66.7%) received antibiotics. Antibiotic use was significantly associated with markers of disease severity, including hypoxia (p = 0.009), tachypnea (p = 0.015), dyspnea (p < 0.001), and hospital admission (p < 0.001). Only 11 (23%) had suspected or confirmed bacterial co-infections. ConclusionA substantial proportion of RSV-positive children received antibiotics. These findings underscore the importance of antimicrobial stewardship programs, rapid diagnostics, and preventive interventions, such as maternal RSV vaccination. Strengthening diagnostic capacity and clinical decision-making in pediatric care is crucial for reducing inappropriate antibiotic use and addressing antimicrobial resistance in low-resource settings.

IntroductionHemophagocytic lymphohistiocytosis (HLH) is a serious condition induced by Dengue virus which becomes fatal if not detected early and treated appropriately. So objectives of the present study are to observe the different patterns of presentations, clinical features and outcome of HLH induced by Dengue. MethodsIn this observational study, 14 patients admitted and diagnosed HLH as per diagnostic criteria, were included after informed written consent. Study conducted in a period of six months from 01/07/2025 to 31/12/2025. All patients were followed up till discharge. After collection, all data were analyzed by Microsoft Excel 2010. Ethical clearance was taken from Ethical Review Board of the Medical College. ResultsAmong 14 cases, male were more affected then the female (78.6% VS 21.4%) and majority were in between 20 to 50 years age groups. Clinical data showed, all 14 cases had fever for >7 days, joint pain 3(21.4%), headache 11(78.6%), skin rashes 10(71.4%), retro-orbital pain 2(14.3%), vomiting 11(78.6%),bleeding 10(71.4%), cough 4(28.6%), loose motion 9(64.3%), abdominal pain 7(50.0%), anorexia 2(14.3%), Melaena 2(14.3%), jaundice 4(28.6%) and spleenomegaly 9(64.3%). One(7.1%) case had history of Hypertension. Laboratory data showed different level of Bi or Pancytopenia, high ferritin, high TG, low fibrinogen, raised liver enzymes and low sodium. Dengue RT PCR and serology results showed 8(42.9%) cases were both IG M and Ig G dengue antibody positive, 6 cases were RT PCR positive, 2 cases were IgM and another 4 cases were IgG positive. Outcome of patients revealed, among all 14 cases12(85.8%) patients improved uneventfully and 2 were shifted to ICU where one improved and one died. ConclusionDengue is prevailing for long time and different complications are evolving and HLH is a relatively newer incident among the dengue patients. Infection by different serotypes at different time or multiple dengue serotype infection may be related with HLH and it might be a future subject to explore and to evaluate.

This report describes lethal Nannizziopsis-associated dermatomycosis in a breeding colony of the family Diplodactylidae (Correlophus ciliatus and Rhacodactylus auriculatus). After introducing one gecko from overseas, three with indirect contact history died due to severe skin lesions. Extensive lesions were observed on the toe pads and ventral surface, along with necrotic dermatitis and cellulitis associated with fungi forming hyphae. Subsequently, four geckos developed diarrhea, melena, emaciation, and fungal dermatitis of the toe pads and died. Histopathologically, the fungal morphologies observed in the skin lesions of the seven geckos were consistent, and Nannizziopsis arthrosporioides was isolated and identified in two of them. To our knowledge, this is the first report of a fatal outbreak of N. arthrosporioides in geckos.

Background: The global burden of dengue infection has rising, yet limited data exists on its impact in the Caribbean. We describe the incidence and associates of acute kidney injury in adults and children with dengue at a teaching hospital in Jamaica. Methods: A single-centre retrospective cohort study of admissions with laboratory confirmed dengue infection at University Hospital of the West Indies, Mona Jamaica between January 2023 to November 2024. AKI was defined using Kidney Disease Improving Global Outcomes definitions. Patients were included if aged >1year and had at least 2 creatinine values. Clinical, demographic and laboratory data were abstracted by chart review. Summary statistics were used to describe continuous and categorical data, and logistic regression to determine AKI associations. Stratified analysis was performed by age-group (adults-aged [&ge;] 16, and paediatric-aged <16 years). Results: Analyses included 167 persons, 62% (103) were male, mean age was 26.1{+/-}19.5 years. AKI occurred in 25.8%, 65.1% were KDIGO stage 1. AKI incidence was 30.2% and 18.0% among adults and children respectively. There were 3 in-hospital deaths. People with AKI were older 32{+/-}21.4 vs 24 {+/-}18.4 (p=0.021), and had longer duration of stay [6 vs 4 days (p <0.001)]. Male sex [OR 2.09 (95% CI:0.96-4.59), p=0.064], age per year [OR 1.02 (95% CI:1.01-1.04), p=0.015] symptom duration [OR1.11 (CI 0.99-1.24), p = 0.058], admission bilirubin [OR 1.02 (CI: 1.00-1.04), p = 0.022], NLR [OR 1.09 (CI 1.00-1.18), p = 0.037] were associated with AKI. In adults admission potassium was inversely associated with AKI [OR 0.46 (95% CI 0.21-1.01), p 0.056], while in children admission potassium [OR 3.00 (95% CI 0.88-10.6), p 0.088] was associated with AKI. Conclusion: AKI in dengue hospitalizations is higher than most reports at 25.8%. Targeted public health policy on vector control and early symptom recognition may be needed to improve outcomes.

Background: How post-COVID-19 condition (PCC) differs from post-acute infection syndromes (PAIS) caused by other respiratory viruses remains uncertain. Comparing these conditions may clarify whether post-acute symptoms reflect specific consequences of SARS-CoV-2 infection or broader post-viral mechanisms. Methods: We conducted a systematic review and meta-analysis of cohort studies comparing persistent symptoms or conditions in adults after SARS-CoV-2 infection with those following other acute respiratory viral infections. PubMed, Embase, and Scopus were searched. Random-effects models were used to estimate pooled risks. Results: Among 9,371 records screened, 22 studies were included and 14 contributed to the meta-analysis. Increased risk after SARS-CoV-2 infection was observed for pulmonary embolism, abnormal breathing, fatigue, hemorrhagic stroke, memory loss/brain fog, and palpitations; heart rate abnormalities showed borderline significance. For most other outcomes pooled estimates were inconclusive. Conclusions: Only a subset of outcomes appears more frequent after SARS-CoV-2 infection, suggesting many symptoms attributed to PCC may reflect broader post-viral syndromes.