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Emerging Infectious Diseases

Centers for Disease Control and Prevention (CDC)

Preprints posted in the last 30 days, ranked by how well they match Emerging Infectious Diseases's content profile, based on 103 papers previously published here. The average preprint has a 0.10% match score for this journal, so anything above that is already an above-average fit.

1
Genomic Epidemiology to Investigate the Origins and Zoonotic Implications of Antibiotic-Resistant Escherichia coli on Beef and Lamb Meat Sold by Independent Butchers in Wales

Sealey, J. E.; Peltonen, N.; Llamazares, B.; Moiseienko, Y.; Mounsey, O.; Taylor, J.; Wright, L.; Williams, P.; Avison, M. B.

2026-03-31 infectious diseases 10.64898/2026.03.30.26349758 medRxiv
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Many studies have identified antibiotic resistant (ABR) Escherichia coli on meat. Appropriate hand hygiene and cooking practices should minimise the risk of gastrointestinal colonisation with ABR E. coli found on meat, and the subsequent chance of causing resistant opportunistic extraintestinal infection. There are large gaps in our understanding of the prevalence, origins and zoonotic potential of ABR E. coli found on meat, however, and particularly for meat reared in extensive farming systems. Wales is a devolved nation within the United Kingdom having large populations of extensively-reared sheep and beef cattle. To help address knowledge gaps around ABR E. coli on extensively reared meat, therefore, beef mince and lamb loin/leg steaks/chops were purchased from 50 (beef) and 46 (lamb) independent butchers across Wales. Following enrichment culture, 200 g meat samples were found to be positive for E. coli resistant to amoxicillin (31% positivity), streptomycin (28%), spectinomycin (29%), amoxicillin-clavulanate (11%), 3rd generation cephalosporins (2%) and fluoroquinolones (5%). Phylogenetic analysis confirmed that Welsh lamb meat ABR E. coli isolates (n=79) are more closely related to those found in faecal samples collected around sheep (n=352) than around beef cattle (n=361) on Welsh farms. This suggests that faecal contamination at or around slaughter is their primary origin. We found no closely related meat/infection clones (<20 SNPs distant and the same antibiotic resistance genes) when comparing ABR E. coli from Welsh meat (n=92) and those causing extraintestinal infections in people (n=2387) in an English region bordering Wales. We conclude, therefore, that the wider zoonotic implications of finding ABR E coli on beef and lamb meat sold at independent butchers in Wales are small.

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Molecular Identification and Characterization of mobatvirus (Hantaviridae) in Lao PDR

Vanhnollat, C.; Dimitrova, K.; Vachouaxiong, L.; Audet, J.; Somlor, S.; Medina, S. J.; Bounmany, P.; Lakeomany, K.; Vungkyly, V.; Wong, G.; Xayaphet, V.; Paphaphanh, P.; Theppangna, W.; Douangboubpha, B.; Vongphayloth, K.; Safronetz, D.; Buchy, P.

2026-04-07 microbiology 10.64898/2026.04.06.713848 medRxiv
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Hantavirids, specifically the members within the genus Orthohantavirus, represent a significant global public health threat, with bat-associated lineages challenging traditional reservoir paradigms. To investigate the genetic diversity of hantavirids in Southeast Asia, we conducted an expanded surveillance program in Lao PDR from May 2023 to October 2025 in bat populations and wild animals from local wet markets. Using molecular screening and deep sequencing to characterize hantavirids from bat populations and wild animals from local wet markets, we identified 20 positive samples across four bat species, recovering coding-complete genomes for multiple novel variants. Phylogenetic analysis confirmed that these viruses form a monophyletic group within Mobatvirus, resolving into two major subclades. The first subclade clustered with Quezon and Robina viruses found in fruit-eating bats. The second subclade further split into two lineages corresponding to Thakrong and Xuan Son viruses, which are associated with trident and leaf-nosed bats, respectively. Despite the strong host specificity observed, the detection of these viruses in a wet market, a critical interface for human-wildlife contact, indicates a potential zoonotic risk. These findings significantly expand the known diversity of mobatviruses in Laos and highlight the urgent need for serological surveillance in at-risk human populations to assess the potential for spillover.

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Diagnostic Accuracy and Potential Resource Savings of Pooled Sputum Testing with Xpert MTB/RIF Ultra for Tuberculosis among adults in Vietnam: A Cross-Sectional Study

Nguyen, H. T.; Codlin, A. J.; Vo, L.; Nguyen, N.; Forse, R.; Dang, H.; Nguyen, L. H.; Hoa, N. B.; Dinh, L. V.; Doan, H. T.; Nguyen Van, H.; Creswell, J.; Garg, T.; Cubas Atienzar, A. I.; Byrne, R. L.; Iem, V.; Squire, B.; Forsman, L. D.; Wingfield, T.

2026-04-01 infectious diseases 10.64898/2026.03.31.26349825 medRxiv
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Objectives: A pooled testing algorithm for tuberculosis (TB), in which sputum specimens from multiple individuals are tested in pools with individual testing of positive pools, can optimise diagnostic resources. This study evaluated the diagnostic accuracy and cartridge savings of pooled testing with the Xpert MTB/RIF Ultra assay (Xpert Ultra) relative to individual Xpert Ultra testing. Methods: We conducted a cross sectional study among 2,396 adults (aged above 15 years) with presumptive TB enrolled between July 2024 and February 2025, through facility based case finding (FBCF) and community based case finding (CBCF). Participants submitted two sputum specimens. The first underwent individual Xpert Ultra testing; remnant specimens were combined into four specimen pools and tested again with Xpert-Ultra. The second specimen was used to inoculate liquid culture (BACTEC MGIT). Data were used to simulate an up-front pooled testing strategy; sensitivity and specificity of this approach was estimated against culture, and cartridge use was compared with individual Xpert-Ultra testing. Results: Of 2,396 participants, 395 (16.5%) had a positive Xpert Ultra and/or culture, including 360/912 (39.5%) in FBCF and 35/1484 (2.4%) in CBCF. The pooled testing approach had sensitivity of 82.4% (95% confidence interval [CI], 77.9; 86.3) and specificity of 98.5% (97.8; 99.0) compared to culture, with lower sensitivity than individual Xpert-Ultra testing (86.5%, 82.4; 89.9) but high specificity (98.1%, 97.4; 98.7). Sensitivity of pooled testing was lower in CBCF (59.1%, 36.4; 79.3) than in FBCF (84.0%, 79.5;87%), whereas cartridge savings were greater in CBCF (69.1% vs 9.6%). The pooling strategy reduced Xpert-Ultra cartridge use by 46.5%, saving USD 14,447. Conclusions: Pooled Xpert-Ultra testing among adults appears resource-efficient for TB screening in Vietnam. As sensitivity is lower compared to individual Xpert Ultra testing, particularly for paucibacillary disease, these losses should be carefully weighed against gains in affordability and expand access to molecular testing. Careful, context-specific implementation is essential to maximise programmatic benefit while minimising missed persons with TB.

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One Health genomics of Acinetobacter baumannii reveals sector-specific lineages and permeable ecological barriers

Plantade, J.; Escobar, C.; Godeux, A.-S.; Poire, L.; Andre, A.; Deromelaere, V.; Cassier, P.; Rasigade, J.-P.; Nazaret, S.; Coluzzi, C.; Venner, S.; Laaberki, M.-H.; Charpentier, X.

2026-04-11 infectious diseases 10.64898/2026.04.09.26350516 medRxiv
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Acinetobacter baumannii is a major cause of severe hospital-acquired infections, with a steadily increasing global prevalence driven by a few clinically adapted lineages. Animals and natural environments also harbor A. baumannii populations, but assessing their connections to clinical lineages is limited by sparse genomic data and a lack of integrated sampling. We conducted a local One Health genomic epidemiology study, sampling, isolating, sequencing, and characterizing several hundred A. baumannii isolates from clinical, animal, and environmental contexts. Within a geographically restricted area, we recovered several globally distributed clinical lineages (international clones, ICs), as well as livestock- and environment-associated lineages shared across Europe, highlighting widespread dissemination beyond clinical settings. Isolates closely related to the emerging clinical lineage IC11 were found in livestock, but no other clinically associated lineages were detected outside clinical contexts. Among these, the epidemic superlineage IC2 was identified in both human and veterinary clinical settings, indicating that similar practices in human and animal medicine select for closely related opportunistic pathogens. We found that hospitals host distinct, antibiotic-sensitive endemic populations capable of causing infection. These populations belong to a diversifying clade spanning clinical and environmental contexts and carry a high load of insertion sequences. Strong plasmid conservation further suggests frequent horizontal gene transfer across ecological compartments. Overall, A. baumannii comprises diverse, context-adapted lineages with a high potential for global spread. Although intercontext transmission appears limited, plasmids may overcome these ecological barriers. Our findings underscore the need for integrated One Health surveillance to better understand transmission pathways and limit the emergence of clinically adapted strains.

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Comparative Evaluation of CLIA and ELISA Serological Assays for HSV-1 IgG with Western Blot Confirmation in a Clinical Cohort

Issa, F.; Trad, F.; Zein, N.; Abunasser, S.; Nizamuddin, P. B.; Salameh, I.; Ayoub, H.; Al-Abbadi, B.; Al-Hiary, M.; Abou-Nouar, Z.; Al-Subeihi, O.; Al-Zubi, Y.; Al-Manaseer, A.; Al-Jaloudi, A.; Nasrallah, D.; Younes, S.; Younes, N.; Abdallah, M.; Pieri, M.; Nicolai, E.; YASSINE, H. M.; Abu-Raddad, L. J.; Nasrallah, G.

2026-04-15 infectious diseases 10.64898/2026.04.14.26350849 medRxiv
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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.

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Tongue swab Xpert MTB/RIF Ultra testing for tuberculosis in adolescents: a cross-sectional study of diagnostic accuracy and acceptability

MacLean, E. L.; Ma, T. T.; Chuong, L. H.; Minh, K. H.; Hoddinott, G.; Pham, Y. N.; Tiep, H. T.; Nguyen, T.-A.; Fox, G.; Nguyen, N. T.

2026-04-25 infectious diseases 10.64898/2026.04.17.26351119 medRxiv
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Introduction Improved diagnostics are needed for people at risk of tuberculosis, especially adolescents. Tongue swab (TS) molecular testing has emerged as a promising strategy for tuberculosis diagnosis. We evaluated diagnostic accuracy and acceptability of Xpert MTB/RIF Ultra (Xpert) using TS samples for tuberculosis detection among adolescents. Methods We conducted a cross-sectional diagnostic accuracy study with consecutive recruitment in Vietnam. Adolescents aged 10-19 who were recommended to undergo investigation for tuberculosis and had not received tuberculosis treatment in the past years were eligible. Participants provided TS and sputum samples and completed a structured survey regarding sampling experiences. TS was tested on Xpert, with sputum tested on Xpert and liquid culture. We utilised a composite reference standard of a positive result on sputum Xpert or sputum culture to define disease status. Sensitivity, specificity, and diagnostic yield were calculated for TS Xpert. Results From July to December 2025, we enrolled 225 adolescents from Can Tho and An Giang provinces in southern Vietnam. Fewer than half (96/225, 43%) the participants exhibited a tuberculosis -like symptom, and the majority (157/225, 70%) were close contacts of a person recently diagnosed with tuberculosis. TS were collected from all adolescents, while 116 (52%) could provide mucopurulent sputum. Tuberculosis prevalence was relatively low (12/225, 5.3%). TS Xpert sensitivity (90% CI) and specificity (90% CI) were 58.3% (35.6, 78.0) and 99.5% (97.9, 99.9), respectively. Diagnostic yield among all diagnosed was 58.3% (7/12). TS sampling was highly acceptable to adolescents; the short time and simplicity of collecting TS were considered favourably. Conclusions The sensitivity and diagnostic yield of TS Xpert was relatively low among adolescents recommended for tuberculosis investigation, which includes asymptomatic individuals who may not provide high quality sputum. Specificity was excellent, and everyone could provide a TS. TS high acceptability indicates it remains a promising sample for diagnostic algorithms.

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Quantitative interferon gamma responses to Mycobacterium tuberculosis in a community-based survey of adolescents and adults in Blantyre, Malawi

Phiri, M. D.; Rickman, H. M.; Mandalasi, C.; Chirambo, A.; Jambo, K. C.; Mwandumba, H. C.; Corbett, E. L.; Horton, K. C.; Henrion, M. Y. R.; MacPherson, P.

2026-04-04 infectious diseases 10.64898/2026.04.02.26349931 medRxiv
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Background: Binary interpretation of Mycobacterium tuberculosis (Mtb) interferon gamma release assay (IGRA) results discards information about recency of exposure and disease risk. We analysed quantitative IGRA responses to Mtb in a community--based survey to investigate associations with response magnitude and inform understanding of transmission dynamics. Methods: We included QuantiFERON--TB Gold Plus (QFT--Plus) results from 2,895 participants (10--40 years old) in Blantyre, Malawi. Bayesian regression models assessed the probability of a positive response ([&ge;]0.35 IU/mL), response magnitude, and associated factors. We also investigated associations with a TB2-TB1 differential >0.6 IU/mL (proposed to reflect recent transmission), and how hypothetical alternative IGRA positivity thresholds affected inference about age-- and sex--specific transmission. Results: 17.4% (503/2,895) of participants had positive TB1 or TB2 responses at the QFT--Plus positivity threshold (0.35 IU/mL). The distributions of TB1 and TB2 responses, among participants with positive QFT--Plus, were similar across age and sex. A TB2-TB1 differential >0.6 IU/mL occurred in 3.8% (109/2,895) of participants and was not associated with age or sex. However, participants with HIV had reduced odds of TB2-TB1>0.6 IU/mL (adjusted odds ratio 0.37 [0.14--0.93]). At higher hypothetical positivity thresholds, the mean predicted Mtb immunoreactivity prevalence among males exceeded that in females at an earlier age: at 19 years, predicted immunoreactivity prevalence ratios were 0.90 (0.83--0.99) and 1.02 (0.89--1.15) at 0.1 IU/mL and 0.5 IU/mL thresholds, respectively. Conclusions: Quantitative IGRA responses offer information about age-- and sex--specific immunoreactivity and transmission risks that dichotomisation obscures. In high-burden settings, quantitative IGRA responses may clarify Mtb transmission patterns and guide targeted public health strategies.

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SARS-CoV-2 neutralising antibody profiles reveal variant specific antibody dynamics and regional differences in infection histories in Malawi

McCormack, M. J.; Banda, L.; Kasenda, S.; Hughes, E. C.; Crampin, A. C.; Amoah, A. S.; Read, J. M.; Ho, A.; Willett, B. J.; Hay, J. A.

2026-04-17 infectious diseases 10.64898/2026.04.16.26351029 medRxiv
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Serological data provide important insights into SARS-CoV-2 transmission and immunity, particularly in regions with limited routine surveillance such as sub-Saharan Africa. However, antibody waning and boosting following reinfection or vaccination remain poorly characterised, complicating interpretation of serological measurements. Improved understanding of these dynamics is critical for accurate epidemiological inference. Modelling longitudinal serological data provides a means to quantify antibody kinetics and reconstruct infection histories. We analysed 15,679 neutralising antibody (nAb) titres from 1,675 unvaccinated, HIV-uninfected participants in urban (Lilongwe) and rural (Karonga) Malawi (February 2021 - April 2022). NAb titres against ancestral B.1, Beta, Delta, and Omicron (BA.1/BA.2) viruses were measured using an HIV-based SARS-CoV-2 pseudotyped virus neutralisation assay. A multi-level Bayesian model was used to reconstruct infection histories and antibody kinetics. The model identified 429 infections (95% credible interval 417-441), including 39 (9.1%) that had not been identified by traditional seroconversion-based thresholds. Antibody levels waned rapidly, with 48% (0.403-0.560) of the acute boost remaining after three months and only 5% (0.027-0.098) after one year. Pre-Omicron infections generated stronger antibody boosts than Omicron infections. Responses varied, with individuals clustering into low and high responders. Cross-reactive responses extended across substantial antigenic distances - Omicron infections induced broader immunity. Seroincidence was higher in Lilongwe than in Karonga (0.41 vs. 0.27 infections per person per three months), driven by the early 2022 Omicron wave. Reinfections were common, particularly among adults and urban residents. SARS-CoV-2 nAb responses following infection were heterogeneous and declined rapidly. This rapid waning underscores the importance of vaccination for sustained protection, while cross-reactivity suggests only partial immunity from prior variants. Identifying reinfections is essential for understanding transmission and finding populations at higher repeat infection risk, particularly where routine surveillance is limited.

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RNase HII-assisted amplification (RHAM) for rapid point-of-care monkeypox detection

Kamhieh-Milz, J.; Kamhieh-Milz, S.; Schwarz, F.; Michel, J.; Nitsche, A.; Puyskens, A.

2026-04-02 infectious diseases 10.64898/2026.04.01.26349928 medRxiv
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Mpox poses an ongoing global public health threat, with case numbers rising beyond traditionally endemic regions in Central and Western Africa. Rapid detection of the causative agent, the Monkeypox virus (MPXV), is critical for outbreak control, yet laboratory infrastructure and trained personnel remain scarce in many affected areas. Point-of-care molecular diagnostics offer a practical solution by enabling timely testing without specialized equipment or elaborate nucleic acid extraction. We evaluated the performance of an extraction-free RNase HII-assisted amplification (RHAM) assay for MPXV detection by Pluslife Biotech, a novel isothermal amplification technology providing results in under 30 minutes. The Pluslife RHAM test demonstrated pan-MPXV clade reactivity, detecting all four MPXV clades (Ia, Ib, IIa, IIb) with high analytical sensitivity and no cross-reactivity to other poxviruses or other clinically relevant pathogens. The assay proved compatible with diverse clinical specimen types, including lesion swabs, oropharyngeal swabs, rectal swabs, urine, semen, and wound exudate. As part of routine diagnostics at the German Consultant Laboratory for Poxviruses, in a comprehensive evaluation of 206 clinical specimens against diagnostic real-time PCR, the Pluslife RHAM test achieved a diagnostic sensitivity of 94.2% (95% CI: 85.8-98.4%) and a specificity of 100% (95% CI: 97.3-100%). Notably, samples with higher viral loads (Ct <30) showed 100% sensitivity. Time-to-result correlated significantly with viral load, enabling faster diagnosis in high-viral-load cases. The Pluslife RHAM test represents a practical, sensitive, and rapid point-of-care solution for MPXV detection in resource-limited settings, combining strong analytical performance with operational simplicity to support timely outbreak response and clinical decision-making.

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Pathologies and causes of death in stranded cetaceans in the Canary Islands (2013-2018)

Diaz Santana, P. J.; Arbelo, M.; Diaz-Delgado, J.; Groch, K.; Suarez-Santana, C.; Consoli, F.; Bernaldo de Quiros, Y.; Quesada-Canales, O.; Sierra, E.; Fernandez, A.

2026-04-05 pathology 10.64898/2026.04.01.715953 medRxiv
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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.

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Tracking cross-border transmission of Rwandas successful dominant rifampicin-resistant Mycobacterium tuberculosis clone using genomic markers

Cuella-Martin, I.; Mulders, W.; Keysers, J.; Hakizayezu, F.; Niyompano, H.; Runyambo, D.; de Rijk, W.-B.; Phelan, J.; Mucyo Habimana, Y.; Migambi, P.; Sawadogo, M.; Mambo Muvunyi, C.; C. de Jong, B.; Ngabonziza, J. C. S.; Rigouts, L.; Meehan, C.

2026-03-31 infectious diseases 10.64898/2026.03.29.26349652 medRxiv
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Background: In Rwanda, genomic surveillance identified a dominant multidrug-resistant tuberculosis (MDR-TB) strain, the R3clone, responsible for approximately 70% of rifampicin-resistant TB cases. Its presence beyond Rwanda remains unexplored. Methods: Unique genetic signatures of the R3clone were defined using whole-genome sequencing of MDR-TB isolates from Rwanda. We developed a targeted qPCR assay detecting a clone-specific single-nucleotide polymorphism. With these tools, we screened isolates from neighbouring countries and public genomic repositories. Results: We identified 375 R3clone isolates, including 264 from historical Rwandan collections (1991-2021), 49 from recent Rwandan diagnostic routine (2021-2024), 25 from historical Burundi isolates (2002-2013), and 37 among public repositories from several countries. The R3clone-specific qPCR showed 100% specificity in distinguishing the R3clone from other MTBC (sub-)lineages. Transmission analysis revealed cross-border transmission of the R3clone within the Great Lakes Region. Conclusion: This study comprehensively assesses cross-border transmission of a dominant MDR-TB strain, highlighting the need for coordinated international surveillance.

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A multicopy transposase-targeted qPCR assay for highly sensitive diagnosis of scrub typhus

Kansuwan, M.; Tapaopong, P.; Anakerit, S.; Chotirat, S.; Tran, B. T.; Charunwatthana, P.; Wattanagoon, Y.; Thawornkuno, C.; Leaungwutiwong, P.; Ahantarig, A.; Nguitragool, W.

2026-04-02 infectious diseases 10.64898/2026.04.01.26349932 medRxiv
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Objectives: Scrub typhus, caused by the bacterium Orientia tsutsugamushi, is frequently underdiagnosed due to its non-specific clinical presentation and the frequent absence of eschar. Most molecular diagnostic assays target single-copy genes of O. tsutsugamushi, which can limit diagnostic sensitivity. We aimed to develop an ultra-sensitive quantitative PCR (qPCR) assay targeting a highly repetitive element in O. tsutsugamushi genome. Methodology: We developed a SYBR Green-based qPCR assay (TranScrub) targeting a multicopy transposase gene of O. tsutsugamushi and compared its performance with assays targeting the 56kDa (single-copy) and traD (multicopy) genes. Diagnostic performance was evaluated using clinical specimens and a panel of blood-borne pathogens. The limit of detection (LOD) was estimated using serial dilutions of quantified template. The assay was further applied to dried blood spot (DBS) samples from patients with acute febrile illness of unknown aetiology, with positives confirmed by Oxford Nanopore amplicon sequencing. Results: Targeting the multicopy transposase gene enabled highly sensitive detection of O. tsutsugamushi, outperforming the conventional 56-kDa assay and matching the traD assay. TranScrub achieved a 91% sensitivity (29/32) and 100% specificity (77/77) using blood-derived DNA, with no cross-reactivity. The LOD was 0.024 genome equivalents/L. Among 81 DBS samples from acute febrile patients of unknown aetiology, 6 (7.5%) tested positive, all confirmed by sequencing. Conclusions: The transposase gene represents a novel target that improves molecular detection of scrub typhus. TranScrub enables sensitive and specific detection from both blood and DBS, supporting its use in clinical diagnosis and field surveillance.

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Buffaloed in Brandenburg: Germany's first Brush with Foot-and-Mouth Disease after four Decades of Freedom

Eschbaumer, M.; Staubach, C.; Pfaff, F.; Gethmann, J.; Schulz, K.; Rogoll, L.; Bock, S.; Bock, W.-I.; Schulze, C.; Marquart, R.; Reinhardt, N.; Nickisch, S.; Kakerow, N.; Freter, S.; Rudovsky, A.; Albrecht, K.; Leo, S.; Haarmann, C.; Lenz, S.; Hoffmann, B.; Calvelage, S.; Höper, D.; Zitzow, P.; Breithaupt, A.; Cokcaliskan, C.; Parlak, U.; Karniely, S.; Abdulrasool, L. M. S.; Knowles, N.; Girault, G.; Romey, A.; Bakkali, L.; King, D. P.; Kühn, C.; Sauter-Louis, C.; Beer, M.

2026-03-31 microbiology 10.64898/2026.03.30.713672 medRxiv
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Foot-and-mouth disease (FMD) virus is one of the most feared and most consequential pathogens of livestock worldwide. It can be spread rapidly by the transboundary movement of animals, animal products and byproducts. In January 2025, Germany detected its first FMD outbreak since 1988 in extensively reared water buffalo on a small farm in the state of Brandenburg, directly outside Berlin, the federal capital. Immediate control measures including a standstill for movements of susceptible animals and pre-emptive culling were implemented by the veterinary authorities. Whole-genome sequencing identified the virus as serotype O, topotype ME-SA, lineage SA-2018 and revealed extensive recombination, but cross-neutralization assays suggested good heterologous protection by an O/PanAsia-2 vaccine strain. Epidemiological back-calculation placed the time of virus introduction in late December 2024. Although the entry route remains unresolved, human-associated introduction is most likely. Network analysis revealed minimal farm connectivity, and simulations predicted low potential for onward transmission, which is consistent with the outbreak being ultimately restricted to a single herd. This event underscores the constant and unpredictable risk of introduction of the virus. Early detection through increased awareness and comprehensive differential diagnostics as well as the international collaboration of veterinary services, laboratories and experts are essential in the face of the global presence of FMD.

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SARS-CoV-2 Introductions into Lao PDR Revealed by Genomic Surveillance, 2021-2024

Panapruksachat, S.; Troupin, C.; Souksavanh, M.; Keeratipusana, C.; Vongsouvath, M.; Vongphachanh, S.; Vongsouvath, M.; Phommasone, K.; Somlor, S.; Robinson, M. T.; Chookajorn, T.; Kochakarn, T.; Day, N. P.; Mayxay, M.; Letizia, A. G.; Dubot-Peres, A.; Ashley, E. A.; Buchy, P.; Xangsayarath, P.; Batty, E. M.

2026-04-13 epidemiology 10.64898/2026.04.09.26349480 medRxiv
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We used 2492 whole genome sequences from Laos to investigate the molecular epidemiology of SARS-CoV-2 from 2021 through 2024, covering the major waves of COVID-19 disease in Laos including time periods of travel restrictions and after relaxation of travel across international borders. We identify successive waves of COVID-19 caused by shifts in the dominant lineage, beginning with the Alpha variant in April 2021 and continuing through the Delta and Omicron variants. We quantify a shift from a small number of viral introductions responsible for widespread transmission in early waves to a larger number of introductions for each variant after travel restrictions were lifted, and identify potential routes of introduction into the country. Our study underscores the importance of genomic surveillance to public health responses to characterize viral transmission dynamics during pandemics.

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Animal husbandry and environmental conditions are associated with cefotaxime-resistant Escherichia coli in yard soil in peri-urban Malawi

Budden, E.; Niven, C. G.; Clark, B.; Floess, E.; Chirwa, B.; Matekenya, M.; Cadono, S.; Chavula, J.; Chisamanga, V.; Dzinkambani, A.; Kaponda, C.; Ngondo, N.; Patterson, N.; Symon, S.; Chunga, B. A.; Holm, R. H.; Chigwechokha, P.; de los Reyes, F. L.; Workman, C. L.; Harris, A. R.; Ercumen, A.

2026-03-27 public and global health 10.64898/2026.03.24.26349245 medRxiv
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Soil is an important reservoir for antimicrobial resistance (AMR) and increasingly recognized as a pathogen transmission pathway. While studies have detected AMR in soil in various settings, dominant contributors to domestic soil contamination with antimicrobial-resistant organisms in low-income countries remain unidentified. We conducted a cross-sectional study with 237 households in southern Malawi, specifically peri-urban Bangwe near Blantyre, to identify factors associated with the abundance of cefotaxime-resistant E. coli in yard soil. Enumerators employed structured surveys and sampled 30 cm2 of yard soil per household. We used IDEXX Quanti-Tray/2000 with Colilert-18 and cefotaxime supplement to enumerate the most probable number (MPN) of cefotaxime-resistant E. coli per dry gram of soil. We conducted multivariable regression to assess associations between the abundance of cefotaxime-resistant E. coli and household sanitation, animal ownership and management, child health and antibiotic use, and weather. Of 228 soil samples, 68% harbored cefotaxime-resistant E. coli at a mean of 0.90 log10-MPN/dry gram. Compared to households without animals, households had approximately 0.50-log lower mean cefotaxime-resistant E. coli abundance in soil if animals were enclosed at night and 0.50-log higher abundance if they were not (p-values<0.005). Additionally, samples had approximately 0.70-log lower mean cefotaxime-resistant E. coli abundance if soil was dry at the time of collection and if it came from a household in the top wealth quintile (p-values<0.005). Daytime animal confinement, household sanitation, child health, antibiotic use, rainfall, temperature and ambient humidity were not associated with cefotaxime-resistant E. coli abundance. Findings suggest that animal husbandry and soil moisture had stronger associations with cefotaxime-resistant E. coli in soil compared to sanitation or antibiotic use. These results underscore the importance of a One Health approach and the relevance of domestic animals and environmental factors to AMR in soil. Studies should quantify soilborne AMR exposure and evaluate associations with specific animal management/enclosure practices.

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Genomic Evidence of Multidrug-Resistant Salmonella in Wild Waterbirds from High-Andean Lakes of Ecuador

Reyes, N.; Vinueza-Burgos, C.; Medina-Santana, J.; Ishida, M. L.; Sauders, B. D.; Anchatuna, D.; Luzuriaga-Neira, N.

2026-04-11 microbiology 10.64898/2026.04.09.717384 medRxiv
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Salmonella spp. represents a leading cause of foodborne disease globally. Wild aquatic birds inhabiting ecosystems impacted by human activities may serve as reservoirs and dispersers of Salmonella and antimicrobial resistance genes (ARGs), posing significant public health risks. This study evaluated the prevalence, serovars, resistance genes, and genomic relationships of Salmonella in fecal samples from wild aquatic birds across three high-Andean lakes in Ecuador. Of 134 samples collected from 10 species, five (3.73%) tested positive, all from Yahuarcocha Lake, isolated from Fulica ardesiaca and Phalacrocorax brasilianus. Two serovars were identified: Salmonella Infantis (ST32, n=4) and Salmonella Newport (ST45, n=1). Three S. Infantis isolates exhibited multidrug resistance (MDR), mediated by a pESI-like plasmid carrying resistance genes against beta-lactams, aminoglycosides, tetracyclines, sulfonamides, trimethoprim, fosfomycin, and chloramphenicol. SNP-based phylogenetic analysis revealed low genetic divergence ([&le;]10 SNPs) between wildlife and poultry-associated isolates, indicating a shared transmission network. These findings support a likely spillover from poultry production systems into wild bird populations, and highlight the role of wild aquatic birds as ecological sentinels and potential disseminators of MDR Salmonella across interconnected human, animal, and environmental systems. These results underscore the need to incorporate human, animal, and environmental health factors within a One Health framework.

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Ocular findings in Northern Gannets following an outbreak of high pathogenicity avian influenza (H5N1)

Fontane, C.; Blacklock, B.; Kayes, D.; Parker, J.; Cunningham, E.; Ravenswater, H.; Jeglinski, J. W.; Mackley, E.; Franklin, K. A.; Tapia-Harris, C.; Philbey, A. W.; Wilson, L. A.; Santos, M.; Lane, J. V.

2026-04-18 physiology 10.64898/2026.04.15.718625 medRxiv
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BackgroundDuring 2021-2022, high pathogenicity avian influenza (HPAI) caused mass mortality in wild birds across Europe, with Northern Gannets (Morus bassanus) among the most affected. Following the outbreak, unusual alterations in the species characteristic pale iris were observed in some individuals. MethodsOpportunistically captured gannets on Bass Rock (n=52), selected to represent a range of iris pigmentation, were examined. Slit-lamp biomicroscopy, indirect ophthalmoscopy, rebound tonometry and photography were performed. Iris pigmentation was classified as normal, mottled or black. Eleven birds underwent avian influenza virus (AIV) serology. Histopathology was performed on two eyes. ResultsAbnormal iris pigmentation was found in 74% of adult and immature gannets, with 61% affected bilaterally. Additional signs consistent with uveitis were present in 77% of affected birds. Iris pigmentation abnormalities were positively associated with AIV H5 seropositivity (Fishers exact test, P=0.018). Histopathology from affected eyes showed increased melanin deposition and disorganisation, including loss of a distinct anterior layer of melanocytic cells and hypertrophy of melanocytes within the iris stroma. LimitationsField conditions limited uniform lighting and concurrent serology. ConclusionsIris pigmentation changes were associated with prior HPAI exposure and frequently accompanied by signs of uveitis, suggesting iris alterations may indicate past infection and potential chronic sequelae.

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Establishment of Contextually Appropriate Cut Offs for Orthopoxvirus Serologic Assays in an Mpox-Endemic Setting

Frederick, C.; Merritt, S.; Halbrook, M.; Mukadi, P.; Anta, Y.; Kompany-Kisenzele, J. P.; Tambu, M.; Makangara-Cigolo, J.-C.; Hasivirwe Vakaniaki, E.; Kenye, M.; Lunyanga, L.; Kacita, C.; Kalonji, T.; Kinanga, C.; Linsuke, S.; Hensley, L. E.; Bogoch, I. I.; Shaw, S. Y.; Hoff, N. A.; Mbala-Kingebeni, P.; Rimoin, A. W.; Kindrachuk, J.

2026-04-14 infectious diseases 10.64898/2026.04.10.26350607 medRxiv
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Mpox virus (MPXV) gained increased attention following the declaration of two Public Health Emergencies of International Concern (PHEICs) in 2022 and 2024. The rapid spread of MPXV and the increase in human-to-human transmission highlighted the need for improved diagnostic tools for characterizing infection patterns and transmission dynamics. While PCR is effective for detecting active infections, serological approaches can help identify previous or asymptomatic infections and support retrospective surveillance. However, many serological assays developed during recent outbreaks have not been evaluated in endemic settings such as the Democratic Republic of the Congo (DRC). This study aims to define antigen-specific serological cutoff values to differentiate MPXV-seroreactive individuals from those with other orthopoxvirus (OPXV) exposure or different vaccination histories, specifically for use in the DRC. Here, we analyzed 134 individuals, divided into six distinct cohorts with different exposures. Serum samples were tested using Mesoscale Discovery (MSD) to screen for five MPXV and vaccinia virus (VACV) orthologous antigens: A29L/A27L, A35R/A33R, B6R/B5R, E8L/D8L, and M1R/L1R. Receiver operating characteristic (ROC) analysis identified the best-performing antigens and established seroreactivity cutoff values. A binary composite rule was also evaluated to improve the classification of these results. We identified three MPXV antigens, E8L (cut-off=12.33 AU/mL), A35R (cut-off=5.22 AU/mL), and B6R (cut-off=9.77 AU/mL), that showed the strongest discriminatory performance in the dataset. Collectively, these three antigens form a significant panel that demonstrated clear separation between our mpox survivor cohort and other OPXV-exposed individuals.

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Comparative genome analysis of Corynebacterium ulcerans Japanese isolates revealed domestically and globally diverse geographical distribution of the organism of different types.

Kimura, M.; Tsuyoshi, S.; Kuroda, M.; Senoh, M.; Takahashi, M.; Yamamoto, A.; Iwaki, M.

2026-04-08 microbiology 10.64898/2026.04.05.716610 medRxiv
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Corynebacterium ulcerans is a zoonotic pathogen causing infections in humans that are clinically indistinguishable from those of Corynebacterium diphtheriae. In many developed countries, companion animals such as dogs and cats, are the predominant sources of infection by C. ulcerans, as is the case in Japan. To date, we have collected 106 Japanese clinical and domestic animal isolates of C. ulcerans and a recently branched species, C. ramonii, most of which were isolated from humans and cats. In the present study, we performed a comparative analysis of whole-genome sequences from Japanese isolates and 597 published isolates from Japan and outside Japan. The Japanese isolates were divided into two distinct lineages: C. ulcerans and C. ramonii. The MLST types of C. ulcerans Japanese isolates exhibited a unique distribution pattern, with one major type (ST337) accounting for 69.0%, which is very rare in Europe. In addition to Japan, distinct MLST pattern compositions were observed across geographically distinct regions. The sequence types were associated with lysogenized prophage types that encode the diphtheria toxin (tox) gene and were partially associated with toxin production under certain conditions. Through SNV analysis, transmission from animals to humans has been suggested in some clinical cases. Significance StatementBy analyzing more than 700 genomes, we demonstrate striking geographic differences in multilocus sequence types and toxigenic prophage compositions between Japanese and European isolates. Notably, we identify the predominance of sequence type ST337 in Japan, a lineage that is rare in Europe, and show that prophage types encoding the diphtheria toxin are closely associated with specific sequence types and toxin production phenotypes. Our single-nucleotide variant analyses further provide genomic evidence supporting zoonotic transmission between companion animals and humans in several clinical cases.

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Reluctant Owners and Unwanted Guns: Exploring Motivations for Relinquishing Firearms at Gun Buybacks in Michigan

Humphreys, D. K.; Giraldi, D. M.; Solomon, S.; Trumbull, E.; Wiebe, D. J.

2026-03-31 public and global health 10.64898/2026.03.29.26349650 medRxiv
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Background: Firearms are frequently transferred through inheritance and other non-purchase pathways, leaving many individuals in possession of unwanted guns and limited options for safe disposal. This study examined the characteristics and motivations of individuals relinquishing firearms at community gun buyback and destruction events in Michigan to inform understanding of firearm divestment and disposal pathways. Methods: We conducted an explanatory sequential mixed-methods study of six faith-based gun buyback and destruction events held in southeastern Michigan between June and October 2024. Quantitative surveys (n = 109) captured participant demographics and firearm characteristics. Follow-up qualitative interviews (n = 7) explored participants' experiences and motivations using inductive-deductive thematic analysis. Results: Across six events, 151 individuals relinquished 318 firearms, most of which were handguns. Nearly one-third of participants disposed of firearms on behalf of others, and two-thirds of personally owned guns had been obtained through non-purchase transfers, most commonly inheritance. Participants frequently expressed anxiety about storing unwanted firearms and relief after safe disposal. The most common motivations were concern about misuse (59%) and fear of theft (54%). Interviews identified five intersecting themes: inheritance and unwanted firearms, safety and family protection, evolving views on ownership, barriers to legal disposal, and emotional relief and closure after relinquishment. Conclusions: Many individuals become firearm owners through inheritance or other non-purchase transfers rather than intentional acquisition. Their experiences reveal that unwanted firearms can generate sustained unease and moral responsibility, motivating voluntary divestment when safe, non-punitive options are available. Results: Across six events, 151 individuals relinquished 318 firearms, most of which were handguns. Nearly one-third of participants disposed of firearms on behalf of others, and two-thirds of personally owned guns had been obtained through non-purchase transfers, most commonly inheritance. Participants frequently expressed anxiety about storing unwanted firearms and relief after safe disposal. The most common motivations were concern about misuse (59%) and fear of theft (54%). Interviews identified five intersecting themes: inheritance and unwanted firearms, safety and family protection, evolving views on ownership, barriers to legal disposal, and emotional relief and closure after relinquishment. Conclusions: Many individuals become firearm owners through inheritance or other non-purchase transfers rather than intentional acquisition. Their experiences reveal that unwanted firearms can generate sustained unease and moral responsibility, motivating voluntary divestment when safe, non-punitive options are available.