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Keywords = virus infection technology

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19 pages, 1243 KB  
Review
Host Cell Virus Interactions: Molecular Mechanisms, Immune Modulation, Viral Pathogenesis, and Emerging Therapeutic Targets
by Awadh Alanazi, Mohamed N. Ibrahim, Eman Fawzy El Azab and Mohamed A. Elithy
Viruses 2026, 18(1), 125; https://doi.org/10.3390/v18010125 - 18 Jan 2026
Viewed by 81
Abstract
Host–virus relationships regulate every phase of viral infection and critically influence course of illness and the effectiveness of treatment. Viruses utilize host receptors, intracellular trafficking routes, metabolic programs, and immunological signaling networks to introduce infection, while host cells use innate and adaptive immune [...] Read more.
Host–virus relationships regulate every phase of viral infection and critically influence course of illness and the effectiveness of treatment. Viruses utilize host receptors, intracellular trafficking routes, metabolic programs, and immunological signaling networks to introduce infection, while host cells use innate and adaptive immune responses that both limit viral replication and, in certain situations, cause tissue damage. Given the fast viral evolution and drug resistance linked to virus-directed therapy, there is growing proof that these host-dependent mechanisms are appealing and underutilized targets for antiviral treatment. Recent developments in single-cell technology, proteomics, and functional genomics have made it possible to systematically identify host dependency and restriction factors shared by different viral families, exposing common molecular vulnerabilities that might be targeted therapeutically. This review integrates current knowledge of virus–host interplay via a translational lens, highlighting processes that directly guide the formation of host-directed antivirals and immune-regulating treatments. We emphasize host processes involved in viral entry, replication, and immune signaling that have shown therapeutic significance, while illustrating the difficulties of balancing antiviral effectiveness with immunopathology. By framing host–virus interactions through a therapeutic lens, this review attempts to offer a targeted and translationally relevant viewpoint for next-generation antiviral research. Full article
(This article belongs to the Special Issue Host Cell-Virus Interaction, 4th Edition)
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34 pages, 3567 KB  
Review
Nanobiosensors: A Potential Tool to Decipher the Nexus Between SARS-CoV-2 Infection and Gut Dysbiosis
by Atul Kumar Tiwari, Munesh Kumar Gupta, Siddhartha Kumar Mishra, Ramovatar Meena, Fernando Patolsky and Roger J. Narayan
Sensors 2026, 26(2), 616; https://doi.org/10.3390/s26020616 - 16 Jan 2026
Viewed by 119
Abstract
The emergence of SARS-CoV-2 posed a great global threat and emphasized the urgent need for diagnostic tools that are rapid, reliable, sensitive and capable of real-time monitoring of SARS-CoV-2 infections. Recent investigations have identified a potential connection between SARS-CoV-2 infection and gut dysbiosis, [...] Read more.
The emergence of SARS-CoV-2 posed a great global threat and emphasized the urgent need for diagnostic tools that are rapid, reliable, sensitive and capable of real-time monitoring of SARS-CoV-2 infections. Recent investigations have identified a potential connection between SARS-CoV-2 infection and gut dysbiosis, highlighting the sophisticated interplay between the virus and the host microbiome. This review article discusses the eminence of nanobiosensors, as state-of-the-art tools, to investigate and clarify the connection between SARS-CoV-2 pathogenesis and gut microbiome imbalance. Nanobiosensors are uniquely advantageous owing to their sensitivity, selectivity, specificity, and reliable monitoring capabilities, making them well-suited for identifying both viral particles and microbial markers in biological samples. We explored a range of nanobiosensor platforms and their potential use for concurrently monitoring the gut dysbiosis induced by different pathological conditions. Additionally, we explore how advanced sensing technologies can shed light on the mechanisms driving virus-induced dysbiosis, and the implications for disease progression and patient outcomes. The integration of nanobiosensors with microfluidic devices and artificial intelligence algorithms has also been explored, highlighting the potential of developing point-of-care diagnostic tools that provide comprehensive insights into both viral infection and gut health. Utilizing nanotechnology, scientists and healthcare professionals may gain a more profound insight into the complex interaction dynamics between SARS-CoV-2 infection and the gut microenvironment. This could pave the way for enhanced diagnostic and prognostic approaches, treatment courses, and patient care for COVID-19. Full article
(This article belongs to the Special Issue Feature Review Papers in the Biomedical Sensors Section)
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15 pages, 1782 KB  
Article
Impact of Meteorological Conditions on the Bird Cherry–Oat Aphid (Rhopalosiphum padi L.) Flights Recorded by Johnson Suction Traps
by Kamila Roik, Sandra Małas, Paweł Trzciński and Jan Bocianowski
Agriculture 2026, 16(2), 152; https://doi.org/10.3390/agriculture16020152 - 7 Jan 2026
Viewed by 318
Abstract
Due to its abundance, bird cherry–oat aphid is the most important vector in Poland of the complex of viruses causing barley yellow dwarf virus (BYDV). These viruses infect all cereals. During the growing season, cereal plants are exposed to many species of agrophages, [...] Read more.
Due to its abundance, bird cherry–oat aphid is the most important vector in Poland of the complex of viruses causing barley yellow dwarf virus (BYDV). These viruses infect all cereals. During the growing season, cereal plants are exposed to many species of agrophages, which can limit their growth, development and yield. As observed for many years, global warming contributes to changes in the development of many organisms. Aphids (Aphidoidea), which are among the most important pests of agricultural crops, respond very dynamically to these changes. Under favorable conditions, their populations can increase several-fold within a few days. The bird cherry–oat aphid (Rhopalosiphum padi L.) is a dioecious species that undergoes a seasonal host shift during its life cycle. Its primary hosts are trees and shrubs (Prunus padus L.), while secondary hosts include cereals and various grass species. R. padi feeds directly on bird cherry tree, reducing its ornamental value, and on cereals, where it contributes to yields losses. The species can also damage plants indirectly by transmitting harmful viruses. Indirect damage is generally more serious than direct feeding injury. Monitoring aphid flights with a Johnson suction trap (JST) is useful for plant protection, which enables early detection of their presence in the air and then on cereal crops. To provide early detection of R. padi migrations and to study the dynamics of abundance, flights were monitored in 2020–2024 with Johnson suction traps at two localities: Winna Góra (Greater Poland Province) and Sośnicowice (Silesia Province). The aim of the research conducted in 2020–2024 was to study the dynamics of the bird cherry–oat aphid (Rhopalosiphum padi L.) population in relation to meteorological conditions as recorded by a Johnson suction trap. Over five years of research, a total of 129,638 R. padi individuals were captured using a Johnson suction trap at two locations (60,426 in Winna Góra and 69,212 in Sośnicowice). In Winna Góra, the annual counts were as follows: 5766 in 2020, 6498 in 2021, 36,452 in 2022, 5598 in 2023, and 6112 in 2024. In Sośnicowice, the numbers were as follows: 6954 in 2020, 9159 in 2021, 49,120 in 2022, 3855 in 2023, and 124 in 2024. The year 2022 was particularly notable for the exceptionally high abundance of R. padi, especially in the autumn. Monitoring crops for the presence of pests is the basis of integrated plant protection. Climate change, modern cultivation technologies, and increasing restrictions on chemical control are the main factors contributing to the development and spread of aphids. Therefore, measures based on monitoring the level of threat and searching for control solutions are necessary. Full article
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34 pages, 2079 KB  
Review
Propagation of Emerging and Re-Emerging Infectious Disease Pathogens in Africa: The Role of Migratory Birds
by Babatunde Ibrahim Olowu, Maryam Ebunoluwa Zakariya, Abdulhakeem Opeyemi Azeez, Abdullah Adedeji Al-Awal, Kehinde Samuel Adebayo, Nahimah Opeyemi Idris, Halima Idris Muhammad, Blessing Chizaram Ukauwa and Al-Amin Adebare Olojede
Bacteria 2026, 5(1), 2; https://doi.org/10.3390/bacteria5010002 - 4 Jan 2026
Viewed by 322
Abstract
Migratory birds have been implicated in the spread of diverse emerging infectious pathogens, including West Nile virus, Usutu virus, Avian influenza viruses, Salmonella, Campylobacter, antimicrobial-resistant (AMR) bacteria, and antibiotic resistance genes (ARGs). Beyond their roles as vectors and reservoirs, migratory birds [...] Read more.
Migratory birds have been implicated in the spread of diverse emerging infectious pathogens, including West Nile virus, Usutu virus, Avian influenza viruses, Salmonella, Campylobacter, antimicrobial-resistant (AMR) bacteria, and antibiotic resistance genes (ARGs). Beyond their roles as vectors and reservoirs, migratory birds are also susceptible hosts whose own health may be compromised by these infections, reflecting their dual position in the ecology of pathogens. As facilitators of pathogen transmission during their long-distance migrations, often spanning thousands of kilometres and connecting ecosystems across continents, these birds can easily cross-national borders and circumvent traditional biosecurity measures, thereby acting as primary or secondary vectors in the transmission of cross-species diseases among wildlife, livestock, and humans. Africa occupies a pivotal position in global migratory bird networks, yet comprehensive data on pathogen carriage remain limited. Gaps in knowledge of pathogen diversity constrain current surveillance systems, resulting in insufficient genomic monitoring of pathogen evolution and a weak integration of avian ecology with veterinary and human health. These limitations hinder early detection of novel pathogens and reduce the continent’s preparedness to manage outbreaks. Therefore, this review provides a holistic assessment of these challenges by consolidating existing knowledge concerning the pathogens transmitted by migratory birds in Africa, while recognizing the adverse effect of pathogens, which potentiates population decline, extinction, and ecological imbalance. It further advocates for the adoption of a comprehensive One Health-omics approach that not only strengthens surveillance and technological capacity but also prioritizes the protection of avian health as an integral component of ecosystem and public health. Full article
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28 pages, 19507 KB  
Article
Parvovirus B19 and Cellular Transcriptome Dynamics in Differentiating Erythroid Progenitor Cells
by Erika Fasano, Niccolò Guglietta, Federica Bichicchi, Ilaria Gasperini, Elisabetta Manaresi and Giorgio Gallinella
Viruses 2026, 18(1), 39; https://doi.org/10.3390/v18010039 - 25 Dec 2025
Viewed by 433
Abstract
Parvovirus B19 (B19V) is a human ssDNA virus with ample pathogenic potential. It is characterized by a selective tropism for erythroid progenitor cells (EPC), exerting a cytotoxic effect with blockade of erythropoiesis. In our work, we investigated both viral and cellular expression profile [...] Read more.
Parvovirus B19 (B19V) is a human ssDNA virus with ample pathogenic potential. It is characterized by a selective tropism for erythroid progenitor cells (EPC), exerting a cytotoxic effect with blockade of erythropoiesis. In our work, we investigated both viral and cellular expression profile in the course of infection of EPCs cultures via mRNA high throughput sequencing technology (HTS) and a dedicated bioinformatic pipeline, reconstructing both the viral and cellular transcriptome and their variations. A productive infection was confirmed as restricted to EPCs expressing mature differentiation markers and the specific receptor for virus VP1u region. mRNA HTS reconstructed the viral transcriptome in terms of localization and abundance of the different mRNA species, detailing the differential expression profile of B19V among early or late times in the course of infection. Analysis of cellular transcriptome indicated that variation was mainly driven by the cellular differentiation process, with the virus impacting to a lesser level, but still clearly separating infected vs. non-infected profiles. At early times post-infection, variations were typical of cellular sensing of viral infection and aimed at the induction of an antiviral state. At later times in the course of infection, the cellular population showed induction of an inflammatory response, related to TNF and IL-10, and a transition to adaptive immunity with evidence of upregulation of genes involved in MHC-II presentation. This dual-transcriptome analysis on infected EPCs population can lay the ground for future research aimed at a better definition of the pathogenetic mechanisms of B19V. Full article
(This article belongs to the Collection Parvoviridae)
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31 pages, 4963 KB  
Review
Next—Generation Diagnostic Technologies for Dengue Virus Detection: Microfluidics, Biosensing, CRISPR, and AI Approaches
by Salim El Kabbani and Gameel Saleh
Sensors 2026, 26(1), 145; https://doi.org/10.3390/s26010145 - 25 Dec 2025
Viewed by 996
Abstract
Dengue fever remains a major mosquito–borne disease worldwide, with over 400 million infections annually and a high risk of severe complications such as dengue hemorrhagic fever. The disease is prevalent in tropical and subtropical regions, where population density and limited vector control accelerate [...] Read more.
Dengue fever remains a major mosquito–borne disease worldwide, with over 400 million infections annually and a high risk of severe complications such as dengue hemorrhagic fever. The disease is prevalent in tropical and subtropical regions, where population density and limited vector control accelerate transmission, making early and reliable diagnosis essential for outbreak prevention and disease management. Conventional diagnostic methods, including virus isolation, reverse transcription polymerase chain reaction (RT–PCR), enzyme–linked immunosorbent assays (ELISA), and serological testing, are accurate but often constrained by high cost, labor–intensive procedures, centralized laboratory requirements, and delayed turnaround times. This review examines current dengue diagnostic technologies by outlining their working principles, performance characteristics, and practical limitations, with emphasis on key target analytes such as viral RNA; nonstructural protein 1 (NS1), including DENV–2 NS1; and host antibodies. Diagnostic approaches across commonly used biofluids, including whole blood, serum, plasma, and urine, are discussed. Recent advances in biosensing technologies are reviewed, including optical, electrochemical, microwave, microfluidic, and CRISPR–based platforms, along with the integration of artificial intelligence for data analysis and diagnostic enhancement. Overall, this review highlights the need for accurate, scalable, and field–deployable diagnostic solutions to support early dengue detection and reduce the global disease burden. Full article
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13 pages, 819 KB  
Review
Key Challenges in Screening Blood Donors for Hepatitis B Virus
by Maha A. Badawi, Sahar Eldakhakhny, Mohamed Ali, Mazen Badawi, Jaffar Khiariy, Yagoub Bin-Taleb and Salwa I. Hindawi
Viruses 2026, 18(1), 23; https://doi.org/10.3390/v18010023 - 23 Dec 2025
Viewed by 628
Abstract
Technological advancements in blood donor screening have significantly improved blood safety. However, certain testing challenges and limitations continue to face blood banks in donor screening for the hepatitis B virus, resulting in occasional cases of transfusion transmission. These cases are mostly related to [...] Read more.
Technological advancements in blood donor screening have significantly improved blood safety. However, certain testing challenges and limitations continue to face blood banks in donor screening for the hepatitis B virus, resulting in occasional cases of transfusion transmission. These cases are mostly related to donors presenting within the window period and donors with occult hepatitis B infection. There are several other challenges that professionals in transfusion medicine, infectious diseases, gastroenterology, and public health must be aware of. Maintaining the highest test sensitivity is a key parameter for enhancing blood safety, and the review describes current recommendations in this regard, along with relevant advancements. The diversity of viral genotypes and the potential for mutations affecting the surface antigen may negatively affect the performance of both serologic and nucleic acid tests. Serologic tests may also be affected by several interferences, endogenous or exogenous to the sample. A clear understanding of these challenges is necessary to create effective policies and procedures and to properly manage atypical cases. Full article
(This article belongs to the Section General Virology)
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20 pages, 1661 KB  
Article
Structure-Guided Engineering of Protein VP2 from Epizootic Hemorrhagic Disease Virus Maximizes Production and Confers Complete Protection as Subunit Vaccine
by Samuel Jurado, Luis Jiménez-Cabello, María del Carmen Nuñez, Sergio Utrilla-Trigo, Eva Calvo-Pinilla, Iván Mazuecos-Aragonés, José Ramón Gutierrez, Ana Falcón, Javier Ortego and José M. Escribano
Vaccines 2026, 14(1), 7; https://doi.org/10.3390/vaccines14010007 - 20 Dec 2025
Viewed by 477
Abstract
Epizootic hemorrhagic disease (EHD) is an important livestock disease caused by Epizootic hemorrhagic disease virus (EHDV). The recent incursion and wide distribution of EHDV in Europe have increased the need for effective vaccine candidates. Background/Objectives: The VP2 protein of EHDV forms the outer [...] Read more.
Epizootic hemorrhagic disease (EHD) is an important livestock disease caused by Epizootic hemorrhagic disease virus (EHDV). The recent incursion and wide distribution of EHDV in Europe have increased the need for effective vaccine candidates. Background/Objectives: The VP2 protein of EHDV forms the outer capsid layer of the virion and is essential for viral assembly and host cell entry. Owing to its antigenic properties, VP2 represents a major target for vaccine development. However, the recombinant production of VP2 is limited by low stability and poor yields, representing a significant barrier for the generation of safe and effective subunit vaccines. Methods: To overcome these limitations, the VP2 protein from EHDV serotype 8 (EHDV-8) was rationally engineered with targeted modifications at both the amino and carboxyl termini of its coding sequence. Recombinant expression was performed using a baculovirus vector-mediated system in Trichoplusia ni pupae (CrisBio® technology), employed as living biofactories. Results: The engineering of VP2 resulted in up to a tenfold increase in protein yields compared with the wild-type sequence, while maintaining the trimeric structural integrity of the recombinant protein. Both wild-type and engineered VP2 protein variants were formulated and used to immunize IFNAR(−/−) mice, a model susceptible to EHDV infection. Both engineered and wild-type VP2 formulations elicited comparable neutralizing antibody responses in vaccinated animals. Furthermore, immunization with either formulation conferred full protection against lethal EHDV-8 challenge. Conclusions: In this work, we demonstrated that the rational engineering of the VP2 protein significantly improved recombinant expression yields in a baculovirus-based system without compromising structural integrity or immunogenicity. These findings additionally demonstrate the feasibility of producing high-quality VP2 antigens in T. ni pupae using CrisBio® technology and support their potential application in the development of subunit vaccines against EHDV. Full article
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20 pages, 913 KB  
Review
Post-Translational Modifications in Respiratory Virus Infection: Recent Insights into the Development of In Vitro Models
by Emna Ben Khlifa, Alessia Campese, Andrea Corsi, Cristina Bombieri, Maria Grazia Romanelli, Maria Teresa Valenti, Donato Zipeto, Matteo Castelli, Patricia Marie-Jeanne Lievens and Alessandra Ruggiero
Int. J. Mol. Sci. 2025, 26(24), 12174; https://doi.org/10.3390/ijms262412174 - 18 Dec 2025
Viewed by 479
Abstract
Post-translational modifications (PTMs) are crucial chemical alterations occurring on proteins post-synthesis, impacting various cellular processes. During viral infections, PTMs are shown to play a multitude of roles in viral replication, host interaction, and immune evasion. Thus, these modifications can influence infectivity, with direct [...] Read more.
Post-translational modifications (PTMs) are crucial chemical alterations occurring on proteins post-synthesis, impacting various cellular processes. During viral infections, PTMs are shown to play a multitude of roles in viral replication, host interaction, and immune evasion. Thus, these modifications can influence infectivity, with direct impact on the anti-viral host immune responses and potentially viral adaptation across species. This field is still scarcely explored, whilst understanding PTMs is not only important to advance the knowledge of virus pathology but also potentially to provide insights for vaccine development. In this review, we attempt to summarize the latest findings mainly published over the last 10 years, focusing on the roles of PTMs involved in virus infection and anti-viral immune responses, in the context of relevant human respiratory infections: influenza A virus (IAV), respiratory syncytial virus (RSV), and SARS-CoV-2. We decided to concentrate on these three viruses because they currently represent a global health problem due to recurrent outbreaks and pandemic potential. A deeper characterization of the PTMs may help in understanding virus–host interaction with possible implications on curative strategies. Further, we will report on cutting-edge technologies to study in vitro virus infection in different cellular-based systems. In particular, we describe and discuss the application of 2D and 3D lung organoid cell-culture systems as in vitro models to mimic respiratory environments and to study the PTMs in a controlled setting. Finally, we will discuss the importance of PTMs in the context of next-generation vaccine design, especially for their potential role to offer effective protection against respiratory viruses. Full article
(This article belongs to the Special Issue Viral Infections and Immune Responses)
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26 pages, 741 KB  
Review
A Crosstalk Between Periodontal Disease and Human Immunodeficiency Virus: Application of Artificial Intelligence and Machine Learning in Risk Assessment and Diagnosis—A Narrative Review
by Bhavyasri Gaddam, Leela Subhashini C. Alluri, Ihunna Amugo, Lemlem Berta, McKayla Butler, Shania Ferguson, Alexys Ferguson, Ethel Harris, Vladimir Berthaud, Siddharth Pratap, Qingguo Wang, Chethan Sampath, Zaid H. Khoury and Pandu R. Gangula
Dent. J. 2025, 13(12), 603; https://doi.org/10.3390/dj13120603 - 16 Dec 2025
Viewed by 443
Abstract
Periodontal disease (PD) is an inflammatory condition caused by multiple periodontal pathogens, particularly those belonging to the Red Complex. Various risk factors influence the development of PD, including age, sex, socioeconomic status, ethnicity, and underlying health issues. Numerous molecular and cellular processes govern [...] Read more.
Periodontal disease (PD) is an inflammatory condition caused by multiple periodontal pathogens, particularly those belonging to the Red Complex. Various risk factors influence the development of PD, including age, sex, socioeconomic status, ethnicity, and underlying health issues. Numerous molecular and cellular processes govern the inflammatory response, which affects the gums and tooth-supporting structures and ultimately leads to alveolar bone loss. Accumulating evidence suggests that Human Immunodeficiency Virus-1 (HIV-1) infection significantly impacts the initiation and progression of PD. While HIV-1 is treated with antiretroviral therapy, this treatment can also affect the course of periodontal disease and systemic health status. AI/ML and precision medicine integrates genomic and computational data to enable individualized disease prevention and treatment strategies. When applied responsibly, these technologies can assist clinicians in the timely detection of both PD and HIV-1. This review aims to discuss the factors that exacerbate PD and the available therapeutic options for persons living with (PLWH) and without HIV-1. Additionally, we emphasize the need for developing biomarkers for early diagnosis and intervention to manage PD effectively, ultimately improving the quality of life for those living with HIV. Full article
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13 pages, 495 KB  
Review
Advances in Novel Detection Technologies for Occult Hepatitis B Virus Infection: Building an Ultra-Sensitive Barrier for Transfusion Safety
by Meng Yi, Yuwei Hu, Bin Fan, Yiming Pan, Bo Pan, Jue Wang and Zhong Liu
Microorganisms 2025, 13(12), 2821; https://doi.org/10.3390/microorganisms13122821 - 11 Dec 2025
Viewed by 485
Abstract
Occult hepatitis B virus infection (OBI), characterized by extremely low viral loads and the persistent intrahepatic presence of cccDNA, poses a profound challenge to global public health security. With a prevalence ranging from 0.06% to over 15% in different donor populations, OBI maintains [...] Read more.
Occult hepatitis B virus infection (OBI), characterized by extremely low viral loads and the persistent intrahepatic presence of cccDNA, poses a profound challenge to global public health security. With a prevalence ranging from 0.06% to over 15% in different donor populations, OBI maintains a risk of transmission and can progress to hepatocellular carcinoma. Its prevention and control have long been limited by the sensitivity constraints of conventional detection methods, highlighting the urgent need for more sensitive diagnostic innovations. Emerging technologies offer distinct breakthroughs: ddPCR facilitates absolute quantification; CRISPR-Cas systems coupled with isothermal amplification enable rapid, point-of-care testing; third-generation sequencing resolves viral integration and mutations; and nanomaterials enhance the signal detection. This review synthesises advancements in OBI diagnostic technologies and provides a comparative overview of their strengths, limitations, and transfusion safety implications, as well as their potential applications in blood transfusion. Recommendations are also proposed to inform the advancement of OBI risk control in blood transfusion and to guide the development of novel diagnostic technologies, particularly relevant to regions with high HBV endemicity, such as China. Full article
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16 pages, 6867 KB  
Article
A Polycistronic tRNA-amiRNA System Reveals the Antiviral Roles of NbAGO1a/1b/2 Against Soybean mosaic virus Infection
by Wenhua Bao, Danyang Sun, Yan Qiu, Xiaoke Zhao and Hada Wuriyanghan
Plants 2025, 14(24), 3724; https://doi.org/10.3390/plants14243724 - 6 Dec 2025
Viewed by 494
Abstract
RNA interference (RNAi) is a crucial antiviral defense mechanism in plants, where Argonaute (AGO) proteins play a central role. However, the function of AGO proteins in the interaction between Soybean mosaic virus (SMV) and Nicotiana benthamiana remains unclear. In this study, SMV pathogenicity [...] Read more.
RNA interference (RNAi) is a crucial antiviral defense mechanism in plants, where Argonaute (AGO) proteins play a central role. However, the function of AGO proteins in the interaction between Soybean mosaic virus (SMV) and Nicotiana benthamiana remains unclear. In this study, SMV pathogenicity was confirmed using an SMV-GFP infectious clone, with typical symptoms and systemic GFP fluorescence observed 14 days post-inoculation. Real-time quantitative reverse transcription polymerase chain reaction analysis revealed dynamic regulation of multiple NbAGO genes upon infection. Notably, NbAGO1a, NbAGO1b, and NbAGO2 were significantly upregulated and positively correlated with viral accumulation, suggesting their critical roles in antiviral defense. Based on these findings, these three genes were selected as targets for artificial microRNA (amiRNA) silencing. Three amiRNAs were designed for each gene using the Arabidopsis miR1596 backbone, with the most effective sequences exhibiting silencing efficiencies ranging from 75.2% to 98.1%. A polycistronic tRNA-amiRNA (PTA) cassette was constructed using Golden Gate cloning technology to simultaneously target all three genes. Co-infection assays indicated that the PTA cassette enhanced SMV accumulation more effectively than single amiRNAs, as evidenced by increased GFP fluorescence (49.1–60.5%) and pronounced leaf necrosis. The PTA system downregulated the expression of NbAGO1a, NbAGO1b, and NbAGO2 by 18.4–26.7%. Furthermore, silencing NbAGO2 alone resulted in severe necrosis, underscoring its essential role in this antiviral defense mechanism. This study elucidates the importance of NbAGO1a, NbAGO1b, and NbAGO2 in antiviral immunity and demonstrates the utility of the PTA system for efficient multi-gene silencing, offering valuable insights for developing RNAi-based antiviral strategies. Full article
(This article belongs to the Special Issue Advances in Plant Molecular Biology and Gene Function)
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27 pages, 5391 KB  
Review
Pepino Mosaic Virus in Tomato: Challenges, Control Strategies, and Future Prospects for Resistance Breeding
by Marzena Nowakowska, Julia Minicka, Marcin Nowicki, Wojciech Szczechura and Beata Hasiów-Jaroszewska
Int. J. Mol. Sci. 2025, 26(23), 11749; https://doi.org/10.3390/ijms262311749 - 4 Dec 2025
Viewed by 652
Abstract
Pepino mosaic virus (PepMV) is a highly infectious potexvirus that poses a significant threat to tomato cultivation in greenhouses worldwide. The threat posed by this virus is attributed to by its genetic complexity, characterized by the presence of multiple genotypes in circulation, mixed [...] Read more.
Pepino mosaic virus (PepMV) is a highly infectious potexvirus that poses a significant threat to tomato cultivation in greenhouses worldwide. The threat posed by this virus is attributed to by its genetic complexity, characterized by the presence of multiple genotypes in circulation, mixed infections, and ongoing genotype turnover. Surveys of wild Solanum species have identified promising sources of resistance; however, this resistance is often incomplete, manifesting as symptomless, yet virus-positive, plants. When resistance is identified, introgressing of these traits into elite backgrounds is frequently impeded by reproductive barriers and linkage drag. Consequently, there are currently no commercially available cultivars with durable resistance to PepMV. Current control measures rely on stringent hygiene practices, seed health protocols, and the use of mild isolate cross-protection, which can mitigate fruit symptoms when carefully genotype-matched and closely monitored. Looking forward, achieving durable control will likely require host-centered strategies. Loss-of-susceptibility mutations and RNA interference-based approaches have demonstrated strong potential in experimental studies. Future solutions may involve the integration of genome editing with RNA-based technologies, supported by regulatory harmonization and socioeconomic viability considerations. Full article
(This article belongs to the Section Molecular Plant Sciences)
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18 pages, 5768 KB  
Review
Diagnostic Advances and Public Health Challenges for Monkeypox Virus: Clade-Specific Insight and the Urgent Need for Rapid Testing in Africa
by Caroline N. Sambo, Amanda Skepu, Nolwandle P. Nxumalo and Ketlareng L. Polori
Diagnostics 2025, 15(23), 2991; https://doi.org/10.3390/diagnostics15232991 - 25 Nov 2025
Viewed by 810
Abstract
Background: Monkeypox (MPX), caused by the Monkeypox virus (MPOX) of the Orthopoxvirus genus, has re-emerged as a significant global health threat. Once confined to Central and West Africa, the 2022–2025 multi-country outbreaks, predominantly caused by Clade IIb, demonstrated sustained human-to-human transmission and global [...] Read more.
Background: Monkeypox (MPX), caused by the Monkeypox virus (MPOX) of the Orthopoxvirus genus, has re-emerged as a significant global health threat. Once confined to Central and West Africa, the 2022–2025 multi-country outbreaks, predominantly caused by Clade IIb, demonstrated sustained human-to-human transmission and global spread. Objective: This review summarizes current knowledge on MPX virology, epidemiology, clinical presentation, and diagnostic technologies, with a focus on innovations supporting rapid and field-deployable detection in resource-limited settings. Methods: The recent literature (2019–2025), including peer-reviewed studies, WHO and Africa CDC reports, and clinical guidelines, was critically reviewed. Data were synthesized to outline key developments in diagnostic methodologies and surveillance approaches. Results: MPX comprises two genetic clades: Clade I (Congo Basin) and Clade II (West African), which differ in virulence and transmission. Clade IIb is associated with sexual and close-contact transmission during recent outbreaks. Clinical manifestations have shifted from classic disseminated rash to localized anogenital lesions and atypical or subclinical infections. RT-PCR remains the diagnostic gold standard, while emerging assays such as loop-mediated isothermal amplification (LAMP), recombinase polymerase amplification (RPA), and CRISPR/Cas-based platforms show promise for rapid point-of-care (POC) testing. Complementary serological tools, including ELISA and lateral flow assays, enhance surveillance and immune profiling. Conclusions: The resurgence of MPX highlights the urgent need for accessible, sensitive, and specific diagnostic platforms to strengthen surveillance and outbreak control, especially in endemic and resource-constrained regions. Full article
(This article belongs to the Section Diagnostic Microbiology and Infectious Disease)
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14 pages, 1330 KB  
Article
The First CRISPR-Based Therapeutic (SL_1.52) for African Swine Fever Is Effective in Swine
by Naveen Verma, Alison O’Mahony, Roky Mohammad, Dylan Keiser, Craig W. Mosman, Deric Holden, Kristin Starr, Jared Bauer, Bradley Bauer, Roypim Suntisukwattana, Waranya Atthaapa, Angkana Tantituvanont, Dachrit Nilubol and Douglas P. Gladue
Viruses 2025, 17(11), 1504; https://doi.org/10.3390/v17111504 - 14 Nov 2025
Cited by 1 | Viewed by 1327
Abstract
African swine fever virus (ASFV) is a high-consequence pathogen that causes African swine fever (ASF), for which mortality rates can reach 90–100%, with death typically occurring within 14 days. ASF is currently a highly contagious pandemic disease responsible for extensive losses in pig [...] Read more.
African swine fever virus (ASFV) is a high-consequence pathogen that causes African swine fever (ASF), for which mortality rates can reach 90–100%, with death typically occurring within 14 days. ASF is currently a highly contagious pandemic disease responsible for extensive losses in pig production in multiple affected countries suffering from extended outbreaks. While a limited number of vaccines to prevent ASF are in use in south-east Asia, vaccines are not widely available, are only effective against highly homologous strains of ASFV, and must be used prior to an outbreak on a farm. Currently, there is no treatment for ASF and culling affected farms is the only response to outbreaks on farms to try and prevent spreading. CRISPR/Cas systems evolved as an adaptive immune response in bacteria and archaea that function by cleaving and disrupting the genomes of invading bacteriophage pathogens. CRISPR technology has since been leveraged into an array of endonuclease-based systems used for nucleic acid detection, targeting, genomic cleavage, and gene editing, making them particularly well-suited for development as sequence-specific therapeutic modalities. The programmability of CRISPR-based therapeutics offers a compelling new way to rapidly and specifically target pathogenic viral genomes simply by using different targeting guide RNAs (gRNA) as an adaptable antiviral modality. Here, we demonstrate for the first time a specific CRISPR/Cas9 multiplexed gRNA system that targets the African swine fever viral genome, resulting in sequence-specific cleavage, leading to the reduction in the viral load in infected animals, and subsequent recovery from an otherwise lethal dose of ASFV. Moreover, animals that recovered had protective immunity to subsequent homologous ASFV infection. Full article
(This article belongs to the Special Issue Viral Diseases of Livestock and Diagnostics, 2nd Edition)
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