Virus Variants Research Articles

From the "One-Molecule, One-Target, One-Disease" Concept towards Looking for Multi-Target Therapeutics for Treating Non-Polio Enterovirus (NPEV) Infections.

Non-polio enteroviruses (NPEVs), namely coxsackieviruses (CV), echoviruses (E), enteroviruses (EV), and rhinoviruses (RV), are responsible for a wide variety of illnesses. Some infections can progress to life-threatening conditions in children or immunocompromised patients. To date, no treatments have been approved. Several molecules have been evaluated through clinical trials without success. To overcome these failures, the multi-target directed ligand (MTDL) strategy could be applied to tackle enterovirus infections. This work analyzes registered clinical trials involving antiviral drugs to highlight the best candidates and develops filters to apply to a selection for MTDL synthesis. We explicitly stated the methods used to answer the question: which solution can fight NPEVs effectively? We note the originality and relevance of this proposal in relation to the state of the art in the enterovirus-inhibitors field. Several combinations are possible to broaden the antiviral spectrum and potency. We discuss data related to the virus and data related to each LEAD compound identified so far. Overall, this study proposes a perspective on different strategies to overcome issues identified in clinical trials and evaluate the "MTDL" potential to improve the efficacy of drugs, broaden the antiviral targets, possibly reduce the adverse effects, drug design costs and limit the selection of drug-resistant virus variants.

Disulfide Bond Engineering of Soluble ACE2 for Thermal Stability Enhancement.

Although the primary pandemic of SARS-CoV-2 is over, there are concerns about the resurgence of the next wave of related viruses, including a wide range of variant viruses. The soluble ACE2 (sACE2) inhibits the SARS-CoV-2 spike protein ACE2 interaction and has potential as a variant-independent therapeutic against SARS-CoV-2. Here, we introduce novel disulfide bonds in the wild-type sACE2-Fc by structure-guided mutagenesis, aiming to improve its stability. The stability of each mutant was assessed by a thermal shift assay to screen mutants with increased thermal stability. As a result, we identified a mutant sACE2-Fc with a significantly increased melting temperature. X-ray crystal structure determination of the sACE2 mutant confirmed the correct formation of the designed disulfide bond, and there were no significant structural disturbances. We also proved that the thermostable sACE2-Fc preserved the spike protein binding affinity comparable to the wild-type sACE2-Fc in both molecular and cellular environments, suggesting its therapeutic potential.

Circulating dengue virus serotypes, demographics, and epidemiology in the 2023 dengue outbreak in Chittagong, Bangladesh.

Dengue is a serious epidemic for Bangladesh affecting thousands of lives. This study aimed to identify and determine the prevalence of the circulating variants of dengue virus (DENV) and their association with demographics and clinical manifestations among the dengue-infected patients. A total of 711 participants with NS1 antigen positivity were enrolled, followed by viral RNA extraction from the collected blood samples and a multiplex real-time reverse transcription-polymerase chain reaction (RT-PCR) assay to determine the dengue virus serotypes. Of 711 dengue-infected patients, 503 (70.7%) were male. Among different age groups, most of the patients were 21-30 years old (n = 255, 35.9%). The DENV2 (n = 483, 67.9%) serotype was more prevalent than the DENV3 (n = 144, 20.3%) and DENV1 (n = 84, 11.8%). The duration of fever was highest in the DENV-1 patients (4.79 ± 1.84 days) in contrast to DENV-3 (4.48 ± 1.68 days) and DENV-2 (4.33 ± 1.45 days) (P = 0.039). Importantly, five highly populated areas were identified as dengue hotspots in Chittagong metropolitan city. Our results provide crucial insights into the patterns of dengue virus transmission and severity among southern Bangladeshi population, thereby aiding in the development of targeted public health interventions and management strategies to combat future outbreaks.

Antibodies targeting the spike protein of SARS-CoV-2 can be modified to increase their affinity for new virus variants

Antibodies targeting the spike protein of SARS-CoV-2 can be modified to increase their affinity for new virus variants

Fighting Mutations with Mutations: Evolutionarily Adapting a DNA Aptamer for High-Affinity Recognition of Mutated Spike Proteins of SARS-CoV-2.

An on-going challenge with COVID-19, which has huge implications for future pandemics, is the rapid emergence of viral variants that makes diagnostic tools less accurate, calling for rapid identification of recognition elements for detecting new variants caused by mutations. We hypothesize that we can fight mutations of the viruses with mutations of existing recognition elements. We demonstrate this concept via rapidly evolving an existing DNA aptamer originally selected for the spike protein (S-protein) of wildtype SARS-CoV-2 to enhance the interaction with the same protein of the Omicron variants. The new aptamer, MBA5SA1, has acquired 22 mutations within its 40-nucleotide core sequence and improved its binding affinity for the S-proteins of diverse Omicron subvariants by >100-fold compared to its parental aptamer (improved from nanomolar to picomolar affinity). Deep sequencing analysis reveals dynamic competitions among several MBA5SA1 variants in response to increasing selection pressure imposed during in vitro selection, with MBA5SA1 being the final winner of the competition. Additionally, MBA5SA1 was implemented into an enzyme-linked aptamer binding assay (ELABA), which was applied for detecting Omicron variants in the saliva of infected patients. The assay produced a sensitivity of 86.5 % and a specificity of 100 %, which were established with 83 clinical samples.

Application of chimeric antigens to paper-based diagnostics for detection of West Nile virus infections of Crocodylus porosus – A novel animal test case

Laboratory-based diagnostics such as plaque reduction neutralisation tests (PRNT) and ELISA are commonly used to detect seroconversion to flavivirus infections. However, faster, qualitative screening methods are essential for quicker diagnosis and improved patient outcomes. Lateral flow assays (LFAs) offer rapid results (5–15 mins) at the point-of-care, but few commercial flavivirus antibody detection LFAs are available.We developed an LFA using novel chimeric viral antigens produced by genetically modifying the mosquito restricted Binjari virus (BinJV) to display the outer virion proteins of pathogenic viruses like West Nile virus (WNV). The BinJV chimeric platform offers several advantages for diagnostic assay development, including rapid construction of new chimeras in response to emerging viral variants, safe, scalable antigen manufacturing, and structural indistinguishability to the wild-type pathogenic virion.To demonstrate feasibility, we applied the chimeric WNV (BinJV/WNV) antigen to LFA as the capture/test line reagent for detecting seroconversion in crocodilians to WNV – a virus affecting crocodilians across multiple continents. We confirmed the antigenic conservation of the chimera on the LFA detection surface using monoclonal antibodies. Utilising well-characterised sera (n=60) from WNV-seropositive or flavivirus-naive Australian saltwater crocodiles (Crocodylus porosus), the assay exhibited 98.8 % sensitivity and 100 % specificity, with results obtained in under 15 minutes. The LFA also accurately detected seroconversion in animals experimentally infected with WNV.This qualitative screening method can be performed both inside and outside of a laboratory, and the assay design will guide the optimisation of similar tests for detecting vector-borne viral infections in humans and other animals.

Genomic surveillance of SARS-CoV-2 in Russia: insights from the VGARus platform

Introduction. In response to the COVID-19 pandemic in the Russian Federation, comprehensive response measures were taken. One of these measures was the development of a viral genome aggregation platform (VGARus) to monitor virus variability. The aim of this paper is to describe the role of the VGARus platform in tracking genetic variation in SARS-CoV-2. Materials and methods. VGARus utilizes sequencing data and bioinformatics tools to monitor genetic variations in SARS-CoV-2. The viral genomes were aligned using NextClade, which also translated them into amino acids and identified mutations. The viral variability over time was analyzed by counting the number of amino acid changes compared to the reference sequence. Results. The analysis of data within VGARus enabled the identification of new virus variants, contributing to improved diagnostic tests and vaccine development. The platform allowed for the prediction of epidemiologic trends, facilitating a rapid response to changes in the epidemiologic situation. For example, using VGARus, an increase in COVID-19 incidence was accurately predicted in the summer of 2022 and early 2023, which were associated with the emergence of Omicron subvariants BA.5 and XBB. Data from the platform helps validate the effectiveness of primers and DNA probes to ensure high diagnostic accuracy and reduce the risk of false negatives. Conclusion. VGARus demonstrates the growing role of genomic surveillance in combating COVID-19 and improving preparedness for future infectious disease outbreaks. The platform is a powerful tool for generating evidence-based solutions to combat a pandemic and mitigate its health, economic and societal impacts. It provides the ability to promptly obtain information on the epidemiologic situation in a particular region of the Russian Federation, use genomic data for phylogenetic analysis, compare the mutational spectrum of SARS-CoV-2 sequences with foreign samples. VGARus data allow for both retrospective analysis and predictive hypotheses. For example, we can clearly see the dynamics of the change of different virus variants: sequences belonging to the Alpha, Beta, Delta, Omicron lineages and many less common ones, clearly form the upsurges of morbidity, the interaction of which is reflected in the epidemiological picture. It is also currently being expanded to monitor other pathogens, increasing its public health relevance.

Anticipating Viral Challenges: A Perspective on Phytochemicals Against Existing and Emerging Viruses

Abstract: Viral infections impact millions of individuals annually and in 2018, the WHO called for global preparedness to address potential high-mortality pathogens, referred to as "Pathogen X," which can include fungi, viruses, parasites, or prions. The constant evolution of RNA viruses leads to continually changing variants, challenging the effectiveness of vac-cines and drugs. In underserved healthcare regions, plant-based phytochemicals offer prom-ise in combating viral diseases due to their ready availability, proven effectiveness, and low toxicity. Amidst the evolving virus variants and recurring fatal outbreaks, especially in re-source-constrained regions, phytochemicals hold promise as potential anti-infective agents. This review delves into plant-based antivirals, aiming to update plant-derived antiviral com-pounds' status against existing and emerging viruses from 2019 to 2023. The study aimed to identify active components from medicinal plants with IC50 and EC50 values against human-infecting viruses. It utilized in vitro and in vivo methods to predict phytochemical mechanisms and enhance bioavailability. Among the phytochemicals studied as antivirals, Emodin, Quercetin, Myricetin, Resveratrol, and Silymarin demonstrated efficacy against multiple viruses. Notably, certain plant compounds were effective against multiple viruses and could serve as potential antiviral treatments. Overall, the review illustrates that harness-ing plant-derived compounds shows promise in combating current and evolving infectious threats.

Benefits of Repeated SARS-CoV-2 Vaccination and Virus-induced Cross-neutralization Potential in Immunocompromised Transplant Patients and Healthy Individuals.

Current COVID-19 vaccines primarily target the Spike protein of defined virus variants, offering limited protection against emerging variants in immunocompetent individuals. Similarly, protective immunity following natural SARS-CoV-2 infection is variable and of short duration, raising concerns about immunocompromised individuals' vaccination strategies. This prospective multicenter study examined 66 sera from 59 immunocompromised and 451 sera from 215 immunocompetent individuals from different pandemic periods. We establish and validate a live virus-based neutralization assay to determine the virus-inactivating potential against ancestral and current SARS-CoV-2 isolates. Our virus-based neutralization assay demonstrated superior performance over surrogate neutralization assays. We found strong but transient immunity after complete vaccination schemes, with single doses providing minimum neutralization, regardless of vaccine type. Combining vaccination-induced immunity with SARS-CoV-2 infection before or after vaccination yielded higher neutralizing titers than vaccination or infection alone, consistent across both study groups. Additional doses after a full vaccination course restored neutralization levels. Potentially protective SARS-CoV-2 neutralization is reliably induced in immunocompromised individuals by prior attenuation of immunosuppression. First-generation vaccines protect against various SARS-CoV-2 variants in immunocompetent individuals, with effective cross-neutralization demonstrated up to the Delta variant but largely absent for later Omicron variants. Continuous vaccine updates are necessary to address emerging SARS-CoV-2 variants.

Variability of VPU protein in HIV-1 sub-subtype A6 in patients with different stages of HIV infection

The aim of the study: to compare the genetic diversity of the Vpu protein in HIV-1 in the people living with HIV (PLWH) with different stages of the disease.Materials and methods. An analysis was carried out of 259 clinical samples of whole blood from HIV-infected patients with no experience of taking antiretroviral therapy, who were observed at the Center for the Prevention and Control of AIDS and Infectious Diseases, Moscow, Russia. The analysis included the following stages: extraction of proviral DNA, amplification of the region of the virus genome containing the vpu gene, sequencing of amplification products, genotyping, comparison of conservation and amino acid substitutions in the Vpu protein sequences in PLWH with different stages of the disease.Results and discussion. In 255 out of 259 (98.4%) clinical samples, a sub-subtype A6 virus variant was identified. The consensus sequence of the Vpu sub-subtype A6 protein was obtained, which contained 81 amino acids. No significant differences in the conservation of Vpu protein sequences were found between HIV-1 variants obtained from patients with different stages of the disease. Amino acid substitution P3A was significantly more common in PLWH with the second stage of HIV infection.Conclusion. The results obtained highlight the issue of the influence of non-structural proteins of HIV-1 on the course of the disease and indicate directions for possible research in the future.

A New Variant of Avian Encephalomyelitis Virus Associated with Neurologic Signs in Turkey Poults.

Avian encephalomyelitis (AE) is a disease caused by the avian encephalomyelitis virus (AEV) of the genus Tremovirus in the family Picornaviridae. Recently, cases of turkey poults showing neurological signs were submitted to the veterinary diagnostic laboratories at South Dakota State University and the University of Minnesota. The affected birds were showing nervous neurological signs such as tremors, inability to stand, torticollis, and wing drop. Clinical signs were observed by 3 weeks of age. Necropsy of birds revealed no significant gross lesions in the internal organs, including the brain. There was no significant bacterial growth in the brains. Microscopic examination of various sections of the brain revealed multifocal lymphocplasmacytic perivascular cuffs in the cerebellum and cerebral cortex. The brain samples were processed for detection and whole genome sequencing by next-generation sequencing. Three full-length polyprotein sequences (6405 nt) of AEV were assembled. All three sequences shared 99.9-100% nucleotide and 100% amino acid identities with each other. Only 77.7-78.5% of nucleotide and 90.3-92.5% of amino acid identities with AEV field strains and vaccine sequences were available in GenBank. This indicates that a new divergent variant of AEV is circulating in the field and causing AE outbreaks in the Midwest region.

Nanopore sequencing provides snapshots of the genetic variation within salmonid alphavirus-3 (SAV3) during an ongoing infection in Atlantic salmon (Salmo salar) and brown trout (Salmo trutta)

Frequent RNA virus mutations raise concerns about evolving virulent variants. The purpose of this study was to investigate genetic variation in salmonid alphavirus-3 (SAV3) over the course of an experimental infection in Atlantic salmon and brown trout. Atlantic salmon and brown trout parr were infected using a cohabitation challenge, and heart samples were collected for analysis of the SAV3 genome at 2-, 4- and 8-weeks post-challenge. PCR was used to amplify eight overlapping amplicons covering 98.8% of the SAV3 genome. The amplicons were subsequently sequenced using the Nanopore platform. Nanopore sequencing identified a multitude of single nucleotide variants (SNVs) and deletions. The variation was widespread across the SAV3 genome in samples from both species. Mostly, specific SNVs were observed in single fish at some sampling time points, but two relatively frequent (i.e., major) SNVs were observed in two out of four fish within the same experimental group. Two other, less frequent (i.e., minor) SNVs only showed an increase in frequency in brown trout. Nanopore reads were de novo clustered using a 99% sequence identity threshold. For each amplicon, a number of variant clusters were observed that were defined by relatively large deletions. Nonmetric multidimensional scaling analysis integrating the cluster data for eight amplicons indicated that late in infection, SAV3 genomes isolated from brown trout had greater variation than those from Atlantic salmon. The sequencing methods and bioinformatics pipeline presented in this study provide an approach to investigate the composition of genetic diversity during viral infections.

Integrated viral and immune monitoring in a prospective COVID-19 cohort from India.

In this study, we report on longitudinal kinetics of cellular immune subsets following SARS-CoV-2 infection in a cohort of hospitalized individuals and evaluate the interplay of these profiles with infecting viral variants, humoral immunity including neutralizing responses, vaccination history and clinical outcomes. A cohort of 121 SARS-CoV-2 infected individuals exhibiting varying disease states were prospectively evaluated for lymphopenic profiles, anti-viral humoral responses and infecting viral variants for a period of up to 90 days spanning the period, February 2021-January 2022 (2nd and 3rd waves of infection). A total of 51 participants received at least one vaccine dose of indigenous vaccines (Covishield or Covaxin) prior to recruitment. When stratified in terms of mortality, B and NK cells, in contrast to the T cell compartment, did not recover from nadir levels in non-survivors who were largely unvaccinated. No discriminatory signature was identified for non-survivors in terms of anti-NC or anti-S1-RBD IgG CLIA profiles including GenScript S1-RBD assays. Evaluation of sVCAM and sMAdCAM revealed opposing dynamics that correlated with disease severity and convalescence respectively. Viral variant analysis revealed delta and omicron variants to comprise majority of the infections which reflected national transmission kinetics during the period of recruitment. Our results demonstrate the importance of monitoring circulating biomarkers for convalescence as well as mortality in COVID-19 progression. Delta variants of SARS-CoV-2 clearly demonstrated increased pathogenicity and warrants sustained viral surveillance for re-emergence of these strains. Our findings with respect to vaccination advocate for continued vaccine development and administration of COVID-19 vaccines.

Introduction and temporospatial tracing of piscine orthoreovirus-1 (PRV-1) in Norwegian farmed Atlantic salmon (Salmo salar) after local fallowing.

Piscine orthoreovirus-1 (PRV-1) is a prevalent agent in Atlantic salmon (Salmo salar) and the causative agent of heart and skeletal muscle inflammation (HSMI), an important disease in farmed Atlantic salmon. Investigations into the introduction and dissemination routes of PRV-1 in a field setting have been limited. This study aimed to better understand PRV-1 infections and HSMI-associated mortality under field conditions. We tracked introduction and spread of PRV-1 over one production cycle in a geographically isolated region in Norwegian aquaculture. From five sites, a total of 32 virus isolates were sequenced and genogrouped. The results indicated multiple introductions of PRV-1 to the area, but also revealed a high level of genetic homogeneity among the virus variants. The variants differed from that of the previous production cycle at two out of three sites investigated, suggesting that synchronized fallowing can be a useful tool for preventing dissemination of PRV-1 between generations of fish. Exposure to PRV-1 at the freshwater stage was identified as a potential source of introduction. A low level of HSMI-associated mortality was observed at all sites, with the onset of mortality showing some variation across PRV-1 genogroups. However, the study highlighted the complexity of associating viral genogroups with mortality in a field setting. Overall, this study contributes valuable insights into PRV-1 dynamics in a real-world aquaculture setting, offering potential strategies for disease management and prevention.

Genome sequencing of SARS-Co-V-2 reveals mutations including F559I and V781D in S protein and LI123-124L in the nsp6 in 21K and 21L clades.

Contagious and virulent virus variants like B.1.1.529 have complicated the 2019 global COVID-19 pandemic from Wuhan, China. Omicron, with extensive mutations and high transmissibility, is replacing Delta in some regions. Remarkably, Omicron exhibits reduced disease severity and resistance to certain vaccines and treatments. Our research sought to identify Egypt-specific variants of concern (VOCs) and their mutation patterns, aiming to provide critical insights for tailored public health strategies. We also looked at vaccine compatibility with these VOCs, as well as the efficacy of current treatments against new SARS-CoV-2 variants. We collected 103 PCR-confirmed COVID-19 cases from an Egyptian army hospital and used next-generation sequencing technology to sequence the entire viral genome. The viral genome was then assembled and reconstructed. Nextclade tools aided in clade assignment and Phylogenetic analysis, allowing classification, and understanding of these genomes' Phylogenetic relationships. Our findings reveal that the dominant VOCs in Egypt are the 21K clade, mainly Pango lineages BA.1 (34%), BA.1.1 (30.1%), and BA.1.17 (6.8%), and the 21L clade represented by Pango lineages BA.2. We also identified novel mutations, including F559I in the S protein (consistent in the 21K clade), V781D in the S protein (present in > 50% of both 21K and 21L clades), and LI123-124L in the nsp6 gene (found in both 21K and 21L clades). Finally, our research provides important insights into Egypt's evolving COVID-19 landscape, allowing for tailored responses and risk mitigation strategies for emerging variants in the region.

Antibodies utilizing VL6-57 light chains target a convergent cryptic epitope on SARS-CoV-2 spike protein and potentially drive the genesis of Omicron variants

Continued evolution of SARS-CoV-2 generates variants to challenge antibody immunity established by infection and vaccination. A connection between population immunity and genesis of virus variants has long been suggested but its molecular basis remains poorly understood. Here, we identify a class of SARS-CoV-2 neutralizing public antibodies defined by their shared usage of VL6-57 light chains. Although heavy chains of diverse genotypes are utilized, convergent HCDR3 rearrangements have been observed among these public antibodies to cooperate with germline VL6-57 LCDRs to target a convergent epitope defined by RBD residues S371-S373-S375. Antibody repertoire analysis identifies that this class of VL6-57 antibodies is present in SARS-CoV-2-naive individuals and is clonally expanded in most COVID-19 patients. We confirm that Omicron-specific substitutions at S371, S373 and S375 mediate escape of antibodies of the VL6-57 class. These findings support that this class of public antibodies constitutes a potential immune pressure promoting the introduction of S371L/F-S373P-S375F in Omicron variants. The results provide further molecular evidence to support that antigenic evolution of SARS-CoV-2 is driven by antibody mediated population immunity.

COVID-19 and pregnancy: impact, outcomes, and potential therapy

The impact of the new coronavirus COVID-19 infection on the course of pregnancy, maternal and child health is largely unclear and controversial. The presented literature review analyzes changes in the immune, cardiovascular, and endocrine systems in women in physiological pregnancy. The peculiarities of the course of infection caused by SARS-CoV-2 virus in pregnancy, depending on the virus variant, are presented, the issues of infection pathways into immune and non-immune cells, including placental cells, as well as the issues of transplacental transfer of the virus – the key moment of infection of the embryo or fetus, on which the outcome of pregnancy also largely depends, are considered. Promising approaches to COVID-19 therapy are presented: use of inhibitors of transmembrane serine protease 2 (TMPRSS2), furin, angiotensin-converting enzyme (ACE2) and RNA-dependent RNA polymerase, amnion epithelial cells and their exosomes. At the same time, the potential use of mesenchymal stem cells in patients with severe COVID-19 pneumonia is reviewed. Difficulties and questions regarding the use of the above therapeutic methods in pregnant women are highlighted. Aspects of the use of surfactant preparations in newborns at risk of new coronavirus infection are considered.

Replication Kinetics and Infectivity of African Swine Fever Virus (ASFV) Variants with Different Genotypes or Levels of Virulence in Cell Culture Models of Primary Porcine Macrophages

Replication Kinetics and Infectivity of African Swine Fever Virus (ASFV) Variants with Different Genotypes or Levels of Virulence in Cell Culture Models of Primary Porcine Macrophages

Robust SARS-CoV-2-neutralizing antibodies sustained through 6 months post XBB.1.5 mRNA vaccine booster

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-neutralizing antibodies are substantially expanded 1month after a shot of XBB.1.5 monovalent mRNA vaccine (XBB.1.5 MV) booster, but the durability of this response remains unknown. Here, we address this question by performing neutralization assays on four viral variants (D614G, BA.5, XBB.1.5, and JN.1) using sera from participants obtained at ∼1month, ∼3months, and ∼6months post an XBB.1.5 MV booster. Our findings indicate that the resulting neutralizing antibody titers are robust and generally remain at stable levels for the study period, similar to those following XBB infection. Importantly, this durability of neutralizing antibody titers contrasts with the decline observed after a booster of the original monovalent or BA.5 bivalent mRNA vaccine. Our results are in line with the recent national data from the Centers for Disease Control and Prevention, showing that the efficacy against symptomatic SARS-CoV-2 infection is sustained for up to 4months after an XBB.1.5 MV booster.

Molecular characteristic, evolution, and pathogenicity analysis of avian infectious bronchitis virus isolates associated with QX type in China

Infectious bronchitis virus (IBV) is one of the major avian pathogens plaguing the global poultry industry. Although vaccination is the primary preventive measure for IBV infection, the emergence of virus variants with mutations and recombination has resulted in IBV circulating globally, presenting a challenge for IB control. Here, we isolated 3 IBV strains (CZ200515, CZ210840, and CZ211063) from suspected sick chickens vaccinated with IBV live attenuated vaccines (H120, 4/91, or QXL87). Phylogenetic analysis of the S1 gene sequence of the spike (S) revealed that the 3 isolates belonged to the QX-type (GI-19 lineage). Whole genome sequencing and recombination analysis indicated that CZ200515 and CZ210840 contained genetic material from 4/91 and Scyz3 (QX-type), possibly due to recombination between the circulating strain and the 4/91 vaccine strain, while no evidence of recombination was found in CZ211063. Pathogenicity analysis in 1-day-old specific pathogen-free (SPF) chickens demonstrated that all 3 isolates caused severe tissue damage and varying degrees of mortality. Virus cross-neutralization assay revealed decreased antigen relatedness between the isolates and the QX-type vaccine strain (QXL87). Amino acid sequence homology analysis of S1 revealed 5%-6.5% variances between the isolates and QXL87. Analysis of the S1 subunit structure revealed that mutations of amino acid residues in the hypervariable region (HVR) and the neutralizing epitope region resulted in antigenic variation in isolates by changing the antigen conformation. Our data indicate antigenicity variances between QX isolates and QXL87 vaccine strains, potentially resulting in immune evasion occurrences. Overall, these results offer crucial insights into the epidemiology and pathogenicity of QX-type IBV, facilitating improved selection and formulation of vaccines for disease management.