Virus research最新文献

{"title":"Estimating the impact of direct acting antiviral therapy on the prevalence of hepatitis C virus infection using phylogenetics","authors":"Hossain M.S. Sazzad ,&nbsp;Hui Li ,&nbsp;Behzad Hajarizadeh ,&nbsp;Bethany A. Horsburgh ,&nbsp;Jason Grebely ,&nbsp;Gregory J. Dore ,&nbsp;Rowena A. Bull ,&nbsp;Andrew R. Lloyd ,&nbsp;Chaturaka Rodrigo","doi":"10.1016/j.virusres.2025.199566","DOIUrl":"10.1016/j.virusres.2025.199566","url":null,"abstract":"<div><h3>Introduction</h3><div>Australia has provided unrestricted subsidized access to direct-acting antiviral (DAA) treatment for hepatitis C virus (HCV) infection since 2016. Epidemiological surveillance estimates suggest prevalence of chronic HCV infection has declined since 2016, but these estimates are not separated by genotype and may not capture ‘hidden’ infected populations, notably the most marginalized groups affected, including people who inject drugs and people in prison. This study used phylogenetics to assess whether epidemiological estimates of declining HCV prevalence in the prisons of New South Wales, Australia due to DAA scale up could be reproduced.</div></div><div><h3>Method</h3><div>Near-full-length 280 HCV consensus sequences (GT1a: <em>n</em> = 140, GT3a: <em>n</em> = 140) sampled between 2006 – 2019 from two prison-based cohort studies in NSW were used for phylogenetic estimates. These included 110 acute infection sequences (GT1a: <em>n</em> = 48, GT3a: <em>n</em> = 62) which were considered in a separate sensitivity analysis given the differences in virus mutation rates in acute and chronic infection. Changes in the effective population size of infected people for each genotype were explored with BEAST software suite (v1.10) using a coalescent Bayesian skyline approach.</div></div><div><h3>Results</h3><div>Both the main and sensitivity analyses for GT3a showed a reduction in the effective population size with the latter showing a 36 % decline between 2011–2019 which is more concordant with the decline estimated from non-phylogenetic methods. A decline of similar magnitude was not demonstrated for GT1a. Overall, the analyses using acute infection sequences only were closer to the trends of independent epidemiological estimates.</div></div><div><h3>Conclusions</h3><div>An adequately powered Bayesian evolutionary analysis using acute stage infection sequences may reproduce the decline in HCV infections observed by traditional epidemiological methods during DAA scale up.</div></div>","PeriodicalId":23483,"journal":{"name":"Virus research","volume":"355 ","pages":"Article 199566"},"PeriodicalIF":2.5,"publicationDate":"2025-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143744098","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The association of cycle threshold value with clinical features in patients infected with Omicron variant","authors":"Wen Yang ,&nbsp;Tao Tao ,&nbsp;Jianping Zhang ,&nbsp;Yuting Yao ,&nbsp;Min Chen ,&nbsp;Mingming Liu ,&nbsp;Meiying Wu ,&nbsp;Wei Lei","doi":"10.1016/j.virusres.2025.199565","DOIUrl":"10.1016/j.virusres.2025.199565","url":null,"abstract":"<div><div>This study investigated the correlation between epidemiological and clinical characteristics of patients infected with omicron variants and the cycle threshold (Ct value) for RT-PCR detection. The study population consisted of 115 patients with Omicron infection and the most common symptoms included fever (43.5 %), cough (38.3 %) and sore throat (29.6 %). Laboratory abnormalities were mainly lymphopenia, elevated globulins and elevated blood glucose. Univariate analysis found that older age (<em>P</em> &lt; 0.001) and unvaccinated (<em>P</em> = 0.003) were associated with low Ct values (high viral load). Multivariate analysis showed that an elevated monocyte count (OR: 3.556; 95 % CI: 1.330–9.503) was associated with low Ct values, whereas being vaccinated (OR: 0.209; 95 % CI: 0.051–0.854) and lower serum sodium (OR: 0.137; 95 % CI: 0.051–0.367) were negatively associated with low Ct values. Studies have shown that factors such as monocyte count, vaccination status and serum sodium correlate with Ct values, suggesting the potential of Ct values as a clinical predictor, which could also provide a valuable reference for clinical decision-making.</div></div>","PeriodicalId":23483,"journal":{"name":"Virus research","volume":"355 ","pages":"Article 199565"},"PeriodicalIF":2.5,"publicationDate":"2025-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143742581","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification of host specificity determinants in brome mosaic virus for rice infection","authors":"Yifan Zhang,&nbsp;Masanori Kaido ,&nbsp;Akira Mine,&nbsp;Yoshitaka Takano,&nbsp;Kazuyuki Mise","doi":"10.1016/j.virusres.2025.199564","DOIUrl":"10.1016/j.virusres.2025.199564","url":null,"abstract":"<div><div>Brome mosaic virus (BMV) is a tripartite positive-stranded RNA plant virus. The genomic RNA2 encodes the 2a protein, which has conserved RNA-dependent RNA polymerase motifs and is required for viral RNA replication. In this study, we have used two BMV strains, F and KU5, and identified two key amino acid residues, 776R and 784T, in the C-terminal non-conserved region of the 2a protein that are critical for systemic infection of BMV-F in rice. While KU5 strain was not able to systemically infect rice, the KU5 mutant strain with two codon changes for 776R and 784T in the 2a gene gained the ability to establish systemic infection in rice, which affects long-distance movement, but not replication or cell-to-cell movement. Through infection assays of KU5 synonymous mutant strains, we demonstrated that amino acids, rather than RNA sequences or secondary structures, are responsible for viral infectivity in rice. Computer predictions and yeast two-hybrid screening revealed that the C-terminal region of 2a functions as an intrinsically disordered region, capable of interacting with host proteins. These results provide molecular insights into the host specificity of BMV and advance our understanding of RNA virus evolution and host-pathogen interactions.</div></div>","PeriodicalId":23483,"journal":{"name":"Virus research","volume":"355 ","pages":"Article 199564"},"PeriodicalIF":2.5,"publicationDate":"2025-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143731624","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Analysis of polyclonal and monoclonal antibody to the influenza virus nucleoprotein in different oligomeric states","authors":"Mallory L. Myers ,&nbsp;Michael T. Conlon ,&nbsp;John R. Gallagher ,&nbsp;De'Marcus D. Woolfork ,&nbsp;Noah D. Khorrami ,&nbsp;William B. Park ,&nbsp;Regan K. Stradtman-Carvalho ,&nbsp;Audray K. Harris","doi":"10.1016/j.virusres.2025.199563","DOIUrl":"10.1016/j.virusres.2025.199563","url":null,"abstract":"<div><div>Influenza virus nucleoprotein (NP) is one of the most conserved influenza proteins. Both NP antigen and anti-NP antibodies are used as reagents in influenza diagnostic kits, with applications in both clinical practice and influenza zoonotic surveillance programs. Despite this, studies on the biochemical basis of NP diagnostic serology and NP epitopes are not as developed as for hemagglutinin (HA), the fast-evolving antigen which has been the critical component of current influenza vaccines. Here, we characterized the NP serology of mice, ferrets, and human sera and the immunogenic effects of NP antigen presented as different structural complexes. Furthermore, we show that the classical mouse anti-NP mAb HB65 could detect NP in some commercial influenza vaccines. MAb HB65 bound a linear epitope with nanomolar affinity. Our analysis suggests that linear NP epitopes paired with their corresponding characterized detection antibodies could aid in designing and improving diagnostic technologies for influenza viruses.</div></div>","PeriodicalId":23483,"journal":{"name":"Virus research","volume":"355 ","pages":"Article 199563"},"PeriodicalIF":2.5,"publicationDate":"2025-03-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143714585","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Regional dynamics and mechanisms behind SARS-CoV-2 XDV.1 prevalence in Chongqing via genomic surveillance and molecular insights","authors":"Jin Yan ,&nbsp;Fangyuan Liu ,&nbsp;Sihan Hu ,&nbsp;Junyi Pan ,&nbsp;Qi Yan ,&nbsp;Lu Yao ,&nbsp;Huhao Jin ,&nbsp;Xiaofeng Chen ,&nbsp;Jiuhong He","doi":"10.1016/j.virusres.2025.199562","DOIUrl":"10.1016/j.virusres.2025.199562","url":null,"abstract":"<div><div>The evolution of SARS-CoV-2 has led to the emergence of numerous variants driven by genetic mutations and evolutionary pressures, posing significant challenges to public health. Understanding the molecular mechanisms and epidemiological advantages of variants like XDV.1 remains incomplete. This study analyzed SARS-CoV-2 samples collected in Chongqing from January to August 2024 through genomic surveillance and molecular dynamics simulations. Whole-genome sequencing identified dominant variants, and all-atom simulations assessed the effects of key mutations in the receptor-binding domain (RBD) on ACE2 receptor interactions, including changes in binding free energy. Genomic analysis identified XDV.1 as the dominant variant, characterized by RBD mutations L455S and F456L. These mutations disrupted conserved hydrophobic interactions and caused structural rearrangements. Simulations revealed that these changes increased binding free energy (ΔG = -4.57 kcal/mol) but reduced binding affinity compared to BA.2.86 and JN.1. XDV.1 exhibits structural features suggestive of potential immune evasion mechanisms, including conformational shifts and novel hydrogen-bond networks that could interfere with antibody recognition. These observed structural modifications, rather than increased receptor-binding affinity, may contribute to its widespread prevalence, though direct experimental validation of antibody interactions remains to be investigated. These findings offer valuable insights for vaccine development and epidemiological studies, highlighting the importance of interactions between structural and non-structural proteins in variant adaptation.</div></div>","PeriodicalId":23483,"journal":{"name":"Virus research","volume":"355 ","pages":"Article 199562"},"PeriodicalIF":2.5,"publicationDate":"2025-03-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143731580","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Regulation of inflammatory cytokines and activation of PI3K/Akt pathway by Yiqi Jiedu Formula in recurrent Herpes Simplex Keratitis: Experimental and network pharmacology evidence","authors":"Shuyu Xiao ,&nbsp;Wanhong Miao ,&nbsp;Leilei Wang ,&nbsp;Lei Wang ,&nbsp;Sisi Tang ,&nbsp;Huihui Xu ,&nbsp;Ying Yu","doi":"10.1016/j.virusres.2025.199561","DOIUrl":"10.1016/j.virusres.2025.199561","url":null,"abstract":"<div><h3>Objective</h3><div>This study investigates the therapeutic effects of the Yiqi Jiedu (YQJD) formula on Herpes Simplex Keratitis (HSK) induced by herpes simplex virus type 1 (HSV-1) and elucidates its mechanisms of action through experimental and network pharmacology approaches.</div></div><div><h3>Methods</h3><div>Active ingredients of the YQJD formula were identified using UPLC-HRMS. Network pharmacology was employed to predict shared targets between YQJD and HSK, focusing on the PI3K/Akt signaling pathway. Molecular docking was performed to assess the interaction between key ingredients and targets. <em>In vivo</em>, an HSK mouse model was used to evaluate the YQJD formula's impact on corneal lesions and inflammatory factors. <em>In vitro</em>, human corneal epithelial cells (HCECs) were infected with HSV-1 to assess the formula's effect on IL-4 expression.</div></div><div><h3>Results</h3><div>UPLC-HRMS identified 34 compounds in YQJD, with Isovitexin and Formononetin exhibiting high oral bioavailability. Network analysis revealed 97 intersecting targets, implicating the PI3K/Akt pathway in YQJD's mechanism. Molecular docking showed strong affinities between IL-4, IL-6, and YQJD compounds. <em>In vivo</em>, YQJD significantly improved corneal lesions and modulated the expression of IL-4, IL-6, and AKT. <em>In vitro</em>, YQJD-containing serum regulated IL-4 expression in HCECs post-HSV-1 infection.</div></div><div><h3>Conclusion</h3><div>The YQJD formula ameliorates Herpes Simplex Keratitis by regulating inflammatory cytokines and activating the PI3K/Akt pathway, offering a potential therapeutic strategy for HSK.</div></div>","PeriodicalId":23483,"journal":{"name":"Virus research","volume":"355 ","pages":"Article 199561"},"PeriodicalIF":2.5,"publicationDate":"2025-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143693564","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of a pod pepper vein yellows virus-based expression vector for the production of heterologous protein or virus like particles in Nicotiana benthamiana","authors":"Lujia Wang ,&nbsp;Ge Zhang ,&nbsp;Shan Bu,&nbsp;Zina Lin,&nbsp;Jian Wu,&nbsp;Fei Yan,&nbsp;Jiejun Peng","doi":"10.1016/j.virusres.2025.199559","DOIUrl":"10.1016/j.virusres.2025.199559","url":null,"abstract":"<div><div>Plant viruses are emerging as a compelling alternative system for the heterologous production of pharmaceutical proteins, offering advantages in scalability, cost-effectiveness, and biological safety over traditional expression systems. They are increasingly recognized as effective platforms for biomedical applications, frequently used in the expression of human viral proteins and the display of peptides or proteins. The pod pepper vein yellows virus (PoPeVYV), classified within the genus <em>Polerovirus</em> of the <em>Solemoviridae</em> family, can substantially increase viral titers when co-infected with pod pepper vein yellows virus-associated RNA (PoPeVYVaRNA). This mixed infection methodology facilitates the formation of rod-shaped virus-like particles (VLPs), wherein modified green fluorescent protein (mGFP) is fused to the C-terminus of the coat protein (CP) from the pepper mild mottle virus (PMMoV). Notably, the expression of hepatitis B surface antigen (HBsAg) demonstrates a marked preference for plant viruses, allowing for enhanced expression via the PoPeVYV mixed infection system in <em>Nicotiana benthamiana</em>. Consequently, the PoPeVYV-based vector presents a promising alternative for the high-level production of heterologous proteins and rod-shaped VLPs in plants.</div></div>","PeriodicalId":23483,"journal":{"name":"Virus research","volume":"355 ","pages":"Article 199559"},"PeriodicalIF":2.5,"publicationDate":"2025-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143674565","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Combination therapy enhances the antiviral activity of IFN-λ against SARS-CoV-2 and MERS-CoV","authors":"Vahid Rajabali Zadeh ,&nbsp;Jocelyne M. Lew ,&nbsp;M. Atif Zahoor ,&nbsp;Deanna Santer ,&nbsp;Jordan J. Feld ,&nbsp;Darryl Falzarano","doi":"10.1016/j.virusres.2025.199560","DOIUrl":"10.1016/j.virusres.2025.199560","url":null,"abstract":"<div><div>Therapeutic options against pathogenic human coronaviruses remain limited. In a recent clinical trial, we demonstrated the therapeutic efficacy of pegylated-IFN-λ in COVID-19 outpatients. However, the emergence of variants that have the potential to evade IFN-mediated antiviral responses raises concerns regarding the continued efficacy of this approach. In this work, we compared the sensitivity of SARS-CoV-2 variants and MERS-CoV to IFN-λ treatment <em>in vitro</em> and explored the potential of combination therapy with other FDA-authorized or approved antiviral agents. We observed that in contrast to the ancestral strain, all other SARS-CoV-2 lineages showed varying, but increased resistance to IFN-λ treatment, from a 5.7-fold increase in EC₅₀ value for the P.1 strain to a 32.7-fold increase for the B.1.1.7 variant. We further show that combination treatment with remdesivir or nirmatrelvir enhanced the antiviral effect of IFN-λ against both SARS-CoV-2 and MERS-CoV. These findings justify the initiation of further <em>in vivo</em> testing that ultimately can help inform the development of more effective therapeutic guidelines against pathogenic coronaviruses.</div></div>","PeriodicalId":23483,"journal":{"name":"Virus research","volume":"355 ","pages":"Article 199560"},"PeriodicalIF":2.5,"publicationDate":"2025-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143670780","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The epidemiological analysis of respiratory virus infections in Children in Hangzhou from 2019 to 2023","authors":"Xinfeng Zhao ,&nbsp;Xiaoxiao Zhu ,&nbsp;Jie Wang ,&nbsp;Cuiying Ye ,&nbsp;Shiyong Zhao","doi":"10.1016/j.virusres.2025.199558","DOIUrl":"10.1016/j.virusres.2025.199558","url":null,"abstract":"<div><div>To investigate the infection of children with respiratory tract infection in Hangzhou from 2019 to 2023, and to explore the epidemiological characteristics of respiratory tract virus infection before and after the prevalence of novel coronavirus. A retrospective analysis was conducted on oral and pharyngeal swabs from 302,680 children with acute respiratory infections. Colloidal gold-based assays were used to detect viral antigens, including adenovirus (ADV), influenza A (FluA), influenza B (FluB), and respiratory syncytial virus (RSV). The detection rates of different viruses were statistically analyzed, and comparisons were made regarding infection status before and after the COVID-19 pandemic, mixed infections, seasonal variations, and different age groups. Among the 302,680 samples, 65,493 tested positive for respiratory pathogens, resulting in a positive rate of 21.64 %. The highest positive rate was found for FluA single infections (12.77 %), while mixed infections were primarily observed with ADV combined with other respiratory viruses. There was a significant statistical difference in the overall detection rate of respiratory viruses before (2019), during (2020–2022), and after (2023) the COVID-19 pandemic (χ²=18,074.97, <em>P</em> &lt; 0.001). The positivity rates for FluA (χ²=31,866.75, <em>P</em> &lt; 0.001), FluB (χ²=255.407, <em>P</em> &lt; 0.001), RSV (χ²=338.76, <em>P</em> &lt; 0.001), and ADV (χ²=4110.093, <em>P</em> &lt; 0.001) also showed significant differences before, during, and after the pandemic. The epidemic seasons for FluA and FluB were winter and spring, while ADV did not exhibit a distinct seasonal pattern, and RSV had a peak incidence in winter. The highest positivity rate for ADV was observed in children aged 2–5 years (4.33 %), for FluA in the 5–15-year-old group (17.15 %), for FluB in those over 15 years (4.86 %), and for RSV in children aged 0–2 years (4.37 %). There were significant differences in virus infection positive rates across different age groups (χ²=2615.084, <em>P</em> &lt; 0.001). Additionally, the overall positive rate differed significantly by gender (χ²=87.317, <em>P</em> &lt; 0.001).The four common respiratory pathogens exhibit distinct epidemiological features in terms of age and seasonality. The COVID-19 pandemic has altered the epidemiology of respiratory viruses, particularly in terms of the peak seasons of infection. Clinically, attention should be given to the potential for co-infection with multiple pathogens.</div></div>","PeriodicalId":23483,"journal":{"name":"Virus research","volume":"355 ","pages":"Article 199558"},"PeriodicalIF":2.5,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143634349","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Is it time to consider west Nile and Usutu viruses endemic in central Italy?","authors":"Federico Romiti ,&nbsp;Maria Teresa Scicluna ,&nbsp;Francesco Censi ,&nbsp;Florindo Micarelli ,&nbsp;Silvia Puccica ,&nbsp;Andrea Carvelli ,&nbsp;Marcello Giovanni Sala ,&nbsp;Irene Del Lesto ,&nbsp;Riccardo Casini ,&nbsp;Claudio De Liberato ,&nbsp;Silvia Tofani","doi":"10.1016/j.virusres.2025.199557","DOIUrl":"10.1016/j.virusres.2025.199557","url":null,"abstract":"<div><div>West Nile (WNV) and Usutu (USUV) viruses co-circulated in a region of Central Italy (Lazio) in 2018, as evidenced by the detection of WNV in the nervous tissues of symptomatic horses and USUV in blood donors and mosquito pools. To assess whether these viruses were endemic in the region, we analysed: 1) diapausing <em>Culex pipiens</em> mosquitoes collected during the winter seasons 2022–2023 and 2023–2024, 2) <em>Cx. pipiens</em> mosquitoes collected during the adult activity period from April to November in 2022 and 2023 across 4 provinces, and 3) sera from 52 horses and tissues from 537 birds. Field-collected <em>Cx. pipiens</em>, including both diapausing and non-diapausing individuals, were tested in pools for WNV and USUV using real-time RT-PCR. Serum samples from horses were tested with two WNV ELISA assays, IgM and IgG, while bird tissues were tested for both viruses via real-time RT-PCR. A total of 18,834 <em>Cx. pipiens</em> females were collected, including 9,812 mosquitoes during the winter seasons and 9,022 during the adult activity periods. Mosquitoes were tested in 623 pools, with all pools of diapausing mosquitoes testing negative for both viruses and 12 pools of non-diapausing mosquitoes positive to USUV. The WNV IgG positivity of 7 horse sera, which were negative at the beginning of the study period, was not confirmed by the virus neutralization test. All tissue samples were negative for WNV and USUV. Since WNV and USUV were not detected in diapausing mosquitoes, there was no evidence of the two viruses endemicity in the study area.</div></div>","PeriodicalId":23483,"journal":{"name":"Virus research","volume":"355 ","pages":"Article 199557"},"PeriodicalIF":2.5,"publicationDate":"2025-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143621010","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}