Parkinson’s disease (PD) is a complex neurodegenerative disorder characterized by dopaminergic neuronal loss, motor dysfunction, and a range of non-motor symptoms. While the etiology of PD remains elusive, emerging evidence suggests a significant role for latent herpesvirus infections in its pathogenesis. This study investigates the prevalence and viral loads of herpes simplex virus type 1 (HSV-1), varicella-zoster virus (VZV), Epstein-Barr virus (EBV), and cytomegalovirus (CMV) in PD patients compared to age- and sex-matched healthy controls. Using multiplex PCR and quantitative PCR, we demonstrate a higher prevalence of HSV-1 and VZV in PD patients, with their viral loads correlating significantly with disease severity and elevated levels of interleukin-6 (IL-6), a marker of systemic inflammation. Our findings reveal that active herpesvirus infections exacerbate neuroinflammation, potentially accelerating dopaminergic neurodegeneration. While CMV and HSV-2 showed no significant differences, the co-infection of HSV-1 and VZV was associated with more severe non-motor symptoms, such as cognitive decline and depression. These results underscore the potential of targeting herpesvirus reactivation and associated inflammation as a novel therapeutic approach for managing PD. Antiviral therapies and vaccination strategies, particularly for HSV-1 and VZV, warrant further investigation to mitigate PD progression.
A wide range of species contract a haemorrhagic disease caused by Bluetongue virus (BTV), which is transmitted by hematophagous midges of Culicoides species. The virus shows variation in its pathogenicity and virulence between the mammalian and insect hosts, implicating the role of host proteins in its life cycle. NS3, a versatile protein of BTV, performs a major role in viral egress by acting as perforin. Multifunctional roles of NS3 have also been reported in viral pathogenesis, but there are critical gaps in explaining the exact mechanisms of its crosstalk with the host, creating the space for NS3-host interaction studies. In this study, we report the screening of host proteins interacting with NS3 protein of BTV using yeast-two hybrid (Y2H) assay. An ovine cDNA library derived from sheep lung and the truncated NS3 (NS3t) bait were constructed and evaluated for competency parameters. Following stringent screening, 37 putative host interaction partners were identified, of which eight proteins were selected for validation using alternative yeast-based assays; seven of these interactions were confirmed as positive. NAP1L1/NS3 interaction was further verified with GST pull-down, co-immunoprecipitation and mammalian two-hybrid assays. The results indicate that NAP1L1, a protein involved in nucleosome assembly and cell cycle regulation, interacts with the BTV NS3 protein.
Introduction COVID-19 infection causes significant changes in certain hematological markers and increases inflammatory severity, such as NLR, LMR, PLR, and Ferritin. In contrast, the Pfizer-BNT162b2 mRNA-based vaccine does not induce high levels of inflammation.Methods A prospective cohort study, conducted at the University of Diyala from January 2022 to January 2023, assessed the immune response to the Pfizer-BNT162b2 mRNA vaccine in 45 healthy, unvaccinated medical students aged 18 to 23. The study evaluated Complete Blood Count, proinflammatory cytokines, ferritin, and anti-IgG antibodies at the time of the second dose of vaccination and 4 weeks after the second dose of the vaccine. Biomarkers were analyzed using CBC instrumentation, ELISA, and Cobas e411 assays, with ethical approval granted for the study.Results Hemoglobin levels remained stable, while red blood cell (RBC) volume and RDW-CV significantly decreased. RBC indices (MCH, MCHC, HCT) increased, and total leukocyte counts slightly declined with a notable granulocyte rise. The platelet count remained stable; however, the plateletcrit decreased, while MPV and PDW increased. NLR and PLR levels increased significantly, whereas LMR levels decreased. No significant change occurred in PHR, SIRI, or AIRI. Inflammatory markers (TNF-alpha, ferritin, CD256/APRIL) and anti-spike IgG levels significantly increased after 1 month. Anti-spike IgG levels showed no strong correlations with SIRI, AIRI, PLR, PHR, or MLR, though significant intercorrelations were found among hematological ratios (e.g., PLR with PHR, SIRI, AIRI; LMR with AIRI).Conclusion Pfizer-BNT162b2 mRNA vaccine had a significant impact on the organized redistribution of immune cells.
In this study, a combination of bioinformatics tools was employed to design a multi-epitope peptide-based vaccine against the Crimean-Congo hemorrhagic fever virus (CCHFV). The vaccine construct incorporates immunodominant epitopes derived from the envelopment polyprotein and the RNA-dependent RNA polymerase (L protein) of the Nigerian strain of CCHFV. Two potent adjuvants were included in the design to enhance immunogenicity. The structural and immunological characteristics of the vaccine were thoroughly evaluated. Both secondary and tertiary (3D) structure predictions were performed. The results indicated that the vaccine is antigenic and non-allergenic. Three-dimensional B-cell epitope prediction revealed that 108 residues within the construct are conformational (discontinuous) B-cell epitopes. Molecular docking and dynamic simulations with Toll-like receptors (TLRs) 2, 3, and 8 demonstrated stable interactions between the vaccine and these immune receptors. In silico cloning and mRNA stability analyses suggested that the construct is suitable for expression in Escherichia coli. Additionally, immune simulation results indicated that the vaccine could elicit a robust adaptive immune response following administration. Overall, the designed multi-epitope vaccine candidate demonstrates high structural quality and favorable immunological properties, supporting its potential to induce protective immunity against CCHFV.
The family Amalgaviridae comprises monopartite double-stranded RNA viruses that encode two overlapping open reading frames (ORFs), ORF1 and ORF2. A programmed ribosomal frameshifting (PRF) mechanism facilitates the translation of an ORF1+2p fusion protein. Among the three recognized genera (Amalgavirus, Unirnavirus, and Zybavirus), Zybavirus remains poorly characterized, with only one approved species, Zygosaccharomyces bailii virus Z (ZbV-Z), and a few unclassified proposed members. In this study, we identified four novel zybavirus-like viral genome sequences, tentatively named Zygosaccharomyces bailii virus Z2 (ZbV-Z2), Cryptops hortensis-associated virus Z1 (ChaV-Z1), Drosophila suzukii-associated virus Z1 (DsaV-Z1), and Sand Creek Marshes virus Z1 (SCMV-Z1), from publicly available transcriptome datasets. Phylogenetic analysis placed ZbV-Z2, ChaV-Z1, and DsaV-Z1 in a well-supported clade with ZbV-Z and Xisha Islands zybavirus, supporting their classification within Zybavirus. SCMV-Z1 clustered with seven known viruses in a distinct lineage, which may represent a novel genus within the family Amalgaviridae. Comparative analysis of PRF sites in members of Zybavirus, Amalgavirus, and related clades revealed that UUU_CNN may represent a broader and ancestral consensus +1 PRF motif in this group of viruses. Our study highlights the utility of mining public transcriptome data for novel viral genome discovery and contributes to the refinement of both taxonomic classification and conserved genomic features within this viral family.
Coxsackieviruses (CVs) belong to the genus Enterovirus and family Picornaviridae. Mutants can arise within the replication cycle of RNA viruses. The prototype CVB2 Ohio-1 (CVB2/O) strain adapted to rhabdomyosarcoma (RD) cells induced cytolytic infection and showed three-point mutations in the genome (CVB2/O/RD). The aim of this experimental study was to compare the effect of one or multiple viral mutations in viral protein 1 (VP1) or region 2C of CVB2/O on pathogenesis in two different mouse models. Male A/J and CD1 (10–12 g) mice were infected intraperitoneally with CVB2/O and its mutants or mock infected (control mice). Mice were sacrificed on days 0, 5, 7, 10, and 55 post infection. Blood, heart, and pancreas were collected for virological and histopathological analysis. The presence of viral RNA in the heart and pancreas of infected mice was studied. Different cytokines were detected in the serum. Pathological changes were absent in the hearts of infected mice. Maximum pathological changes were identified in the pancreas of infected A/J mice. Infiltration of pancreatic cells was observed depending on the mouse strain and mutants. CD1 mice were less susceptible to CVB2 infection. CVB2/O/VP1-Q164K mutant induced maximum changes in the pancreas of A/J-infected mice. We suggest that the single altered amino acid in the VP1 protein was related to the virulence factor and was associated with the pathology and presence of viral RNA in the pancreas of infected A/J mice.
Due to the successful eradication of smallpox worldwide and the cessation of smallpox vaccination campaign in 1980, the human population seems to be more susceptible to poxvirus infection. In the last years, an increased detection of zoonotic orthopoxviruses (OPXVs) has also been observed. In particular, in the past 50 years, a high incidence of monkeypox virus (MPXV) disease (MPOX) in both Central and Western Africa was reported. MPXV is not as lethal as variola virus (VARV), the etiological agent of smallpox, but it represents a threat to public health. The global events of MPOX in May 2022, and the ongoing outbreaks in Central and Western Africa in August 2024, have prompted the World Health Organization (WHO) to declare MPXV a Public Health Emergency of International Concern. Preventive vaccination remains the most effective control against MPXV. Smallpox vaccines of the second and third generations have been suggested for high-risk groups, in spite of several limitations, such as some adverse events, reduced immunogenicity, and manufacturing issues. The emerging threat of MPXV highlights the urgent need for the development of vaccines that can effectively control MPOX and potentially prevent diseases caused by other related OPXVs. Our study aims at introducing basic information on the biology of poxviruses, and on factors that may contribute to the reemergence of zoonotic poxviruses. It also summarizes the evolution of vaccinia-based vaccines and strategies that may control and prevent future outbreaks.
Murid gammaherpesvirus 4 (prototype strain MHV-68) is a natural pathogen of murid rodents that is able to transform lymphocytes and induce lymphoproliferative diseases and tumor transformation in their host cells. Considering that this virus can infect and multiply in various human cell lines, we think it could also play an important role in the pathogenesis of some human types of cancer. We investigated and confirmed the presence of the replication and transcription activator rta (ORF 50) from MHV-68 virus in four peripheral blood samples from patients with lymphomas and gliomas.
Grapevine yellow speckle viroid 1 (GYSVd-1) is a widespread subviral pathogen affecting grapevine (Vitis vinifera L.), with potential implications for viticulture. This study investigates the prevalence, genetic variability, intrahost population polymorphism and secondary RNA structure of GYSVd-1 in Slovakia, where its molecular characteristics have been largely unexplored. Leaf samples from 49 grapevine plants across western and southern Slovakia were analyzed using RT-PCR, followed by sequencing and phylogenetic analyses. GYSVd-1 was detected in 73% of the samples, demonstrating high prevalence rates across vineyards in Slovakia. Sequence analysis revealed a high degree of genetic similarity among Slovak isolates, with nucleotide identity ranging from 96.2% to 100%, consistent with low variability reported globally. Phylogenetic analyses showed no specific grouping of Slovak isolates. Intrahost variability, assessed via subcloning of RT-PCR products, demonstrated the presence of random mutations among viroid clones, consistent with quasispecies nature of viroids in their hosts. These findings suggest that both interhost and intrahost viroid populations evolve under similar pressures to maintain structural stability essential for viroid function. Secondary RNA structure analysis of Slovak GYSVd-1 isolates revealed that all unambiguous sequences exhibited the characteristic rod-like conformation. In contrast, a significant proportion of sequences from mixed populations failed to form proper rod-like structures, suggesting that these variants may lack biological viability. Overall, this study unveils the prevalence of GYSVd-1 in Slovak vineyards and provides valuable insights to guide future phylogenetic studies and structural analyses of viroids.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), an enveloped, single-stranded RNA virus, is causative of coronavirus disease 2019 (COVID-19). The viral entry process is facilitated by the highly antigenic spike protein of SARS-CoV-2, which binds to the angiotensin-converting enzyme 2 (ACE2) receptor. However, research involving live SARS-CoV-2 is limited to biosafety level (BSL)-3 facilities, hindering the progress in vaccine and therapeutic development. Pseudotyped viruses have emerged as valuable tools for studying entry inhibitors, fusion inhibitors, and neutralizing assays in BSL-2 facilities. Several modifications to the spike protein have been identified to enhance the yield of pseudotyped viruses. Notably, the removal of the last 19 amino acids from the spike protein and the D614G modification have been shown to increase pseudovirus titers. In this study, we aimed to identify an effective expression vector for pseudotyping by inserting SARS-CoV-2 S Δ19 into the intron-containing pcDNA3.1 vector. The inclusion of the β-globin intron sequence led to a significant augmentation in spike protein expression and a threefold improvement in pseudotyped virus production. Furthermore, we examined whether the woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) could enhance spike protein expression by subcloning the spike protein gene into a lentiviral vector containing WPRE. Our results demonstrated that vectors containing the WPRE exhibited increased spike protein expression compared to vectors lacking the WPRE in HEK293-hACE2 cells. This study demonstrates that the inclusion of a β-globin intron and WPRE enhances the production of SARS-CoV-2 spike protein pseudotyped lentiviruses, a valuable tool for COVID-19 research.
Persistent infection with gammaherpesviruses, such as Epstein-Barr virus and Kaposi’s sarcoma-associated herpesvirus may lead to development of various lymphomas and carcinomas in humans. Murid gammaherpesvirus 4 (MuHV-4, also known as MHV-68) is widely used as a model to study all aspects of gammaherpesvirus pathogenesis. Similar to human gammaherpesviruses, MuHV-4 is known for its variability, as evidenced by the existence of multiple strains/variants that differ from each other in primary sequence and some pathogenetic features. Detailed knowledge of the dependence of pathogenic properties on genome structure and sequence would help to better understand the complex host-pathogen interactions. The main aim of this study was to analyze how various strains/variants of MuHV-4, mainly MHV-72 and MHV-4556 induce latency and reactivation. In this study, we characterized the ability of MHV-72 and MHV-4556 to establish latency in spleen, lungs and thymus of immunocompetent mice after intranasal infection. Moreover, we described the potential of these strains to reactivate ex vivo using the method of tissue explantation and stimulation with inhibitor of histone deacetylases, trichostatin A. Our results clearly demonstrate that both MHV-72 and MHV-4556 exhibit a significant deficit in the establishment of deep latency in spleen. Moreover, MHV-72 has a reduced ability to establish latency in lungs as well as thymus. Regarding reactivation, we found that both MHV-72 and MHV-4556 have a significant deficit in ex vivo reactivation from latency in lungs as well as thymus in comparison to MHV-68. Moreover, MHV-72 also possessed a deficit in the ex vivo reactivation from spleen. In contrast, MHV-4556 was able to respond to the trichostatin A-mediated reactivation stimuli by increasing the DNA copy number but had a defect in the release from tissue during the early and deep latency in spleen. Our results contribute to molecular characterization of differences among the MuHV-4 strains in the ability to establish latency and reactivation and could also serve to better understanding of strain-dependent variances among human gammaherpesviruses.
The epidemic of Monkeypox virus (MPXV), an emerging zoonotic Orthopoxvirus, in beginning OF May 2022, has drawn global attention owing to its increasingly reported cases by the World Health Organization (WHO) in multiple countries. Due to absence of any validated treatments for MPXV infections, the preventive strategies hold significant importance. The current study proposes potential vaccine targets against MPXV by deploying immunoinformatic method. The monkeypox virus encodes 190 different proteins. These proteins were shortlisted on the basis of antigenicity, surface expression, allergenicity, and toxicity. The shortlisted ten proteins were subjected to identification of B- and T-cell epitopes using IEDB, ABCpred, NetMHCpan4.0, and NetMHCIIpan4.0. All the epitopes were further screened for antigenicity, allergenicity, and toxicity properties using VaxiJen, Allertop, and Toxinpred server. All the epitopes have 97.5% worldwide population coverage. Overlapping B-cell, CTL and HTL epitopes were used to design three vaccine construct using suitable linkers and three different adjuvants. Molecular docking was performed for all the three vaccine constructs with TLR4. Based on docking scores and physicochemical profile the vaccine construct V1 with beta defensin adjuvant was selected for further analysis. In silico cloning of V1 into pBAD-DEST49 vector showed maximum expression in bacterial system. Immune simulations also confirmed the high immune responses of V1 within the host cell. The study results may lead towards identification of clinically effective vaccines against MPXV with better safety and potency profile.
Some plant RNA viruses in the family Betaflexiviridae encode a nucleic acid-binding protein (NABP) that facilitates infection by suppressing the host RNA silencing response. Previously, no members of the genus Tepovirus within this family were known to possess a NABP homolog. In this study, we identified the genome sequences of 21 novel Betaflexiviridae viruses: 17 represent new members of Tepovirus, and four may be founding members of a new genus closely related to the genus Vitivirus. Notably, five of these newly identified tepoviruses contain a NABP-like open reading frame (ORF). Sequence comparison and phylogenetic analysis of NABP homologs suggest these tepoviruses independently acquired a NABP-like ORF from diverse sources. The identification of 17 novel viruses substantially enhances our understanding of the genetic diversity within the genus Tepovirus. This study further highlights the role of recombination in the genome evolution and diversity of Betaflexiviridae.
Tick-borne encephalitis virus (TBEV, Flaviviridae), a small enveloped flavivirus with an unsegmented positive-stranded RNA genome, is the most prominent member of the mammalian group of tick-borne flaviviruses. TBEV, originally isolated in 1937, is identified as Orthoflavivirus encephalitidis now. TBEV causes the most important arboviral disease of the human central nervous system (CNS) in Europe and Northeast Asia. It is transmitted to hosts primarily by ticks of the genus Ixodes and Dermacentor, but can also be acquired by ingestion of infected unpasteurized dairy products. Approximately one-third of all human TBEV infections are associated with severe clinical neurological disease. The remaining two-thirds are asymptomatic or present with mild clinical symptoms. In hosts, TBEV tend to induce different types of immune effector mechanisms. Components of innate immunity - natural killer cells, complement proteins, macrophages and dendritic cells usually provide rapid and intense protection in the acute phase of infectious diseases. In turn, cell-mediated immunity provided by T and B lymphocytes plays an important role in virus clearance and protective immunity, and thus influences the outcome of disease. The virus-host relationship is not passive. Therefore, viruses themselves respond actively to host immune defence activities. This is made possible by a number of mechanisms that ensure their escape from the host’s immune surveillance. The aim of this review is to summarize the history of the last 50 years as well as advances in research on the immunology of TBEV, specifically in the Central European area.
More than 20 years after the discovery of the first anelloviruses (AVs), they still remain a mysterious group of viruses. To date, there is no clear understanding of their impact on the host organism, although information is emerging about their participation in various pathologies. Unfortunately, the widespread distribution of AVs makes it difficult to determine their significance. However, it is well known that AVs are an important component of the human virome. We assessed the abundance of AVs in viromes from four individual nasal and pharyngeal samples from children with respiratory symptoms using a metagenomic approach. Three of samples were positive for bocavirus, influenza virus A and respiratory syncytial virus, while one sample was negative for any respiratory viruses in standard PCR diagnosis. AVs were detected in all samples, namely Torque teno mini virus (TTMV). The greatest abundance and diversity of AVs [Torque teno virus (TTV), TTMV and Torque teno midi virus (TTMDV)] were observed in a diseased patient who had an absence of respiratory viruses. It is suggested that an activated immune system to combat the main pathogens, against the background of which the number of AVs decreases, may explain the reduced diversity of anellome in other patients. We also compared the results of the metagenomic workflows and qPCR for major respiratory viruses, which were in agreement. In addition, metagenomic sequencing made it possible to obtain additional data on viral genomes in order to establish their taxonomic identification and characterize individual viral profiles in patients.
BackgroundIndia witnessed three COVID-19 pandemic waves, each with various degrees of severity and clinical signs. The coronavirus strain and immunization status have a significant impact on the severity of COVID-19 infections. The current study intends to evaluate and compare the symptoms, severity, and breakthrough infections in vaccinated and unvaccinated individuals over the three waves of the pandemic.MethodsThis was a retrospective survey study. A Google based questionnaire was used to collect data on demographics, severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection status, re-infections, associated symptoms, severity, and vaccination status over three waves, post-COVID-19 sequelae, and willingness to receive a COVID-19 booster dose in the Indian population. The replies of 3,404 Indian participants were analysed using STATA 11.ResultsEach wave showed a decrease in the number of symptomatic COVID-19 infections. However, fever and loss of smell/taste were identified as the most common symptoms in each wave. Clinical symptoms such as fever, weariness, and shortness of breath were shown to be considerably higher in vaccinated than unvaccinated individuals. The number of SARS-CoV-2 breakthrough infections increased between the second and third waves. Approximately 36.5% of people with protracted COVID-19 had previously received immunization after recovering from a natural COVID-19 illness. Overall, 34.8% of individuals were hesitant to take the COVID-19 booster dose.ConclusionIncreased symptoms in vaccinated individuals during the second wave, emphasizing the potential role of antibody-dependent augmentation. A considerable fraction (36.5%) of those with protracted COVID-19 infections had previously received vaccination after contracting the virus naturally. The fact that vaccine received after COVID-19 infection has been shown to be a risk factor for long-term COVID-19 emphasizes the need for vigilance in this specific subgroup.
BackgroundCurrently, there are approximately 240 million individuals worldwide suffering from chronic hepatitis B infection, yet a definitive cure remains elusive. In the context of pharmaceutical interventions for this patient population, the potential issue of Hepatitis B virus (HBV) reactivation must be taken into consideration. Nutlin-3 is a small molecule of imidazoline analog, which inhibits tumor growth by suppressing the interaction between human double minute 2 (HDM2) and p53. However, whether it affects HBV replication is not clear.MethodsHepG2.2.15 cells were used to assess the impact of Nutlin-3 on HBV replication. The expression of Hepatitis B surface antigen (HBsAg) and Hepatitis B e antigen (HBeAg) was detected by ELISA. The expression of pregenomic RNA (pgRNA) and core-DNA was detected by RT-qPCR. The activation of relevant signaling pathways was assessed through Western blotting and confocal microscopy detection of the expression of associated proteins.ResultsWe found that Nutlin-3 significantly enhances the expression of HBsAg and HBeAg, and it also promotes the expression of pgRNA and Core-DNA. Nutlin-3 promotes HBV replication independent of the HDM2. Instead, it activates the autophagy pathway through p53-mediated mTOR inhibition to promote HBV replication.ConclusionWhen using Nutlin-3 for cancer therapy, it’s important to be aware of the potential side effect of promoting HBV replication in individuals with chronic hepatitis B infection.
Enterovirus 71 (EV71) is a significant causative agent of hand, foot and mouth disease (HFMD). However, the precise mechanism by which EV71 infection leads to alterations in the immune response remain elusive. To address this knowledge gap, we conducted a study where we introduced the Inner Mongolia EV71 C33λ strain into Vero cells, derived from African green monkey kidney cells. Subsequently, we performed RNA sequencing (RNA-Seq) to investigate the changes in the transcriptome of these infected Vero cells. Our primary objective was to establish a foundational understanding that could inform future research on EV71-associated immune factors. In our study, we identified a total of 942 differentially expressed genes (DEGs) in Vero cells infected with Enterovirus 71 (EV71), with 568 gene exhibiting increased expression and 374 gene showing decreased expression. To elucidate the functional implications of these DEGs, we conducted a comprehensive functional enrichment analysis using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. This analysis revealed three genes that were significantly upregulated, which we subsequently validated using reverse transcription polymerase chain reaction technology (RT-qPCR). The RT-qPCR results were in strong agreement with our RNA-Seq data, confirming the reliability of our findings. This study represents the pioneering RNA-Seq analysis that delves into the cellular response of Vero cells to EV71 infection. Our results not only provide a foundational understanding of the molecular changes induced by EV71 but also offer crucial insights into the mechanisms by which EV71 modulates the host immune system. These insights are pivotal for future research endeavors aimed at developing effective therapeutic strategies against EV71 and related pathogens, as well as for understanding the broader implications of viral infections on host immunity.
The global pandemic of COVID-19 caused by SARS-CoV-2 has had a devastating impact. Although many survived the acute effects of the pandemic, a significant number of survivors, including those with only mild symptoms, are now experiencing a prolonged and debilitating post-viral syndrome known as LC/PASC (long COVID/post-acute sequelae of SARS-CoV-2). Typical symptoms of LC/PASC include fatigue, breathlessness, chest pain, impaired cognition, difficulty sleeping, fever and gastrointestinal symptoms. Anxiety and depression can also last for weeks to months and range from mild to disabling. The association between neuropsychiatric symptoms and SARS-CoV-2 infection raises questions about the possible routes of SARS-CoV-2 entry to the central nervous system (CNS) and long-term effects of the virus on the CNS, their molecular basis, and the potential risk of neuronal damage associated with the subsequent development of neurodegenerative diseases.
Enterovirus (EV) infections occur frequently in humans. In some geographical areas they are more common. These viruses cause diseases with varying degrees of severity, from a simple respiratory tract infection to severe diseases. Since EVs include more than 70 serotypes currently circulating in the population, a methodology that detects most of them is needed. ELISA is a rapid, sensitive, and economical diagnostic method for the identification of EV serotypes and can also be used as a retrospective diagnostic tool or in the investigation of outbreaks of infection. Commercial EV-ELISAs often appear and gradually disappear from the market supply. We have used the KTL-510 peptide, a synthetic viral protein of poliovirus VP1, as an antigen in a peptide-based ELISA for the detection of a broader spectrum of anti-EV antibodies. We aimed to design, optimize, and standardize this in-house ELISA with the peptide, and implement the method for routine detection of anti-EV IgG in human sera. For determining the cut-off value, we used 100 patients’ sera which were previously tested negative for IgG antibodies against EVs using a commercial ELISA kit available. We monitored patients’ sera samples sent for serological testing of anti-coxsackievirus antibodies to the National Reference Center for the Identification of Enteric Viruses between 2018–2022. These serum samples were examined using a standard virus neutralization test as well as the newly developed ELISA method.
