Non-enveloped Viruses Research Articles

Rab6a enables BICD2/dynein-mediated trafficking of human papillomavirus from the trans-Golgi network during virus entry.

Human papillomaviruses (HPVs) are small, non-enveloped DNA viruses that cause approximately 5% of human cancer. Like most other DNA viruses, HPV traffics to the nucleus during virus entry to successfully infect cells. We show here that HPV utilizes a cellular enzyme, Rab6a, during virus entry to engage the dynein molecular motor for transport along microtubules. Rab6a is required for complex formation between the HPV L2 capsid protein, dynein, and the dynein adaptor BICD2 in the trans-Golgi network (TGN). This complex is required for transport of the incoming virus out of the TGN as it journeys to the nucleus. Our findings identify potential targets for therapeutic approaches.

Non-lytic spread of poliovirus requires the nonstructural protein 3CD.

Non-enveloped viruses like poliovirus (PV) have evolved the capacity to spread by non-lytic mechanisms. For PV, this mechanism exploits the host secretory autophagy pathway. Virions are selectively incorporated into autophagosomes, double-membrane vesicles that travel to the plasma membrane, fuse, and release single-membrane vesicles containing virions. Loading of cellular cargo into autophagosomes relies on direct or indirect interactions with microtubule-associated protein 1B-light chain 3 (LC3) that are mediated by motifs referred to as LC3-interaction regions (LIRs). We have identified a PV mutant with a severe defect in non-lytic spread. An F-to-Y substitution in a putative LIR of the nonstructural protein 3CD prevented virion incorporation into LC3-positive autophagosomes and virion trafficking to the plasma membrane for release. Using high-angle annular dark-field scanning transmission electron microscopy to monitor PV-induced autophagosome biogenesis, for the first time, we show that virus-induced autophagic signals yield normal autophagosomes, even in the absence of virions. The F-to-Y derivative of PV 3CD was unable to support normal autophagosome biogenesis. Together, these studies make a compelling case for a direct role of a viral nonstructural protein in the formation and loading of the vesicular carriers used for non-lytic spread that may depend on the proper structure, accessibility, and/or dynamics of its LIR. The studies of PV 3CD protein reported here will hopefully provoke a more deliberate look at the presence and function of LIR motifs in viral proteins of viruses known to use autophagy as the basis for non-lytic spread.

Chemical inactivation of two non-enveloped viruses results in distinct thermal unfolding patterns and morphological alterations

BackgroundNon-enveloped viruses, which lack a lipid envelope, display higher resistance to disinfectants, soaps and sanitizers compared to enveloped viruses. The capsids of these viruses are highly stable and symmetric protein shells that resist inactivation by commonly employed virucidal agents. This group of viruses include highly transmissible human pathogens such as Rotavirus, Poliovirus, Foot and Mouth Disease Virus, Norovirus and Adenovirus; thus, devising appropriate strategies for chemical disinfection is essential.ResultsIn this study, we tested a mild, hypoallergenic combination of a denaturant, alcohol, and organic acid (3.2% citric acid, 1% urea and 70% ethanol, pH4) on two representative non-enveloped viruses – Human Adenovirus 5 (HAdV5) and Feline Calicivirus (FCV)– and evaluated the pathways of capsid neutralization using biophysical methods. The conformational shifts in the capsid upon chemical treatment were studied using Differential Scanning Calorimetry (DSC), while the morphological alterations were visualized concurrently using Transmission Electron Microscopy (TEM). We found that while treatment of purified HAdV5 particles with a formulation resulted in thermal instability and, large scale aggregation; similar treatment of FCV particles resulted in complete collapse of the capsids. Further, while individual components of the formulation caused significant damage to the capsids, a synergistic action of the whole formulation was evident against both non-enveloped viruses tested.ConclusionsThe distinct effects of the chemical treatment on the morphology of HAdV5 and FCV suggests that non-enveloped viruses with icosahedral geometry can follow different morphological pathways to inactivation. Synergistic effect of whole formulation is more effective compared to individual components. Molecular level understanding of inactivation pathways may result in the design and development of effective mass-market formulations for rapid neutralization of non-enveloped viruses.

Optimizing RT-qPCR Detection of Enveloped and Non-enveloped Viruses in Wastewater: A Comparative Study in Grenada

Abstract Introduction/Objective Wastewater-based epidemiology (WBE) is an effective methodology to monitor the community spread of pathogens and has successfully been used to identify potential outbreaks. Most standardized methodologies used in WBE for the detection of viruses have been validated for non-enveloped viruses; therefore, there is a need for the development of a standardized protocol for the detection of both enveloped (such as SARS- CoV-2) and non-enveloped viruses (such as Hepatitis A). Wastewater seeded with Pseudomonas syringae bacteriophage (Φ6), an enveloped virus surrogate, and coliphage MS2, a non-enveloped virus surrogate, were used to evaluate the efficiency of four different aluminium flocculation methods. Untreated wastewater spiked with known amounts of surrogates was used as a control for viral recovery. Methods/Case Report Four different viral concentration methods were used. Method 1: AlCl3 flocculation at pH 3.5 followed by centrifugation, resuspension with 10% beef extract and filtered with a 0.22 μm filter. Method 2: AlCl3 flocculation at pH 6.0 followed by room temperature agitation, centrifugation, and resuspension with 3% beef extract. Method 3: Mixed method protocol-AlCl3 flocculation at pH 3.5 followed by room temperature agitation, centrifugation, resuspension with 3% beef extract. Method 4: Mixed method protocol-AlCl3 flocculation at pH 6.0 followed by centrifugation, resuspension with 10% beef extract filtered with a 0.22 μm filter. Viral RNA was extracted after concentration, and reverse-transcription quantitative polymerase chain reaction (RT- qPCR) was performed. Results (if a Case Study enter NA) The average Ct reduction of untreated wastewater compared to spiked was calculated for each method. The highest recovery of both Φ6 and MS2 was obtained using Method 3 (mixed protocol) with a significant difference at 95% confidence in the recovery of Φ6 (p= 0.0029) and MS2 (p= 0.0225). Conclusion Previous results from our lab demonstrate the importance of pH during the flocculation step of viral concentration using different methods of concentration. Here, the results suggest that the increased contact time between virus and the flocs is also critical. This can facilitate the implementation of WBE in Grenada to monitor and track potential viral outbreaks of both enveloped and non-enveloped viruses such as SARS-CoV-2 and Hepatitis A.

Molecular Engineering of Virus Tropism

Engineered viral vectors designed to deliver genetic material to specific targets offer significant potential for disease treatment, safer vaccine development, and the creation of novel biochemical research tools. Viral tropism, the specificity of a virus for infecting a particular host, is often modified in recombinant viruses to achieve precise delivery, minimize off-target effects, enhance transduction efficiency, and improve safety. Key factors influencing tropism include surface protein interactions between the virus and host-cell, the availability of host-cell machinery for viral replication, and the host immune response. This review explores current strategies for modifying the tropism of recombinant viruses by altering their surface proteins. We provide an overview of recent advancements in targeting non-enveloped viruses (adenovirus and adeno-associated virus) and enveloped viruses (retro/lentivirus, Rabies, Vesicular Stomatitis Virus, and Herpesvirus) to specific cell types. Additionally, we discuss approaches, such as rational design, directed evolution, and in silico and machine learning-based methods, for generating novel AAV variants with the desired tropism and the use of chimeric envelope proteins for pseudotyping enveloped viruses. Finally, we highlight the applications of these advancements and discuss the challenges and future directions in engineering viral tropism.

High-Throughput Synthesis and Evaluation of Antiviral Copolymers for Enveloped Respiratory Viruses.

COVID-19 made apparent the devastating impact viral pandemics have had on global health and order. Development of broad-spectrum antivirals to provide early protection upon the inevitable emergence of new viral pandemics is critical. In this work, antiviral polymers are discovered using a combination of high-throughput polymer synthesis and antiviral screening, enabling diverse polymer compositions to be explored. Amphipathic polymers, with ionizable tertiary amine groups, are the most potent antivirals, effective against influenza virus and SARS-CoV-2, with minimal cytotoxicity. It is hypothesized that these polymers interact with the viral membrane as they showed no activity against a nonenveloped virus (rhinovirus). The switchable chemistry of the polymers during endosomal acidification was evaluated using lipid monolayers, indicating that a complex synergy between hydrophobicity and ionization drives polymer-membrane interactions. This new high-throughput methodology can be adapted to continue to engineer the potency of the lead candidates or develop antiviral polymers against other emerging viral classes.

Rapid virus inactivation by nanoparticles-embedded photodynamic surfaces

The persistent threat of viral epidemics poses significant risks to human health, highlighting the urgent need for antiviral surfaces to mitigate viral transmission through bioaerosols and surface contamination. However, there is still a scarcity of readily accessible antiviral coatings to address this critical concern. In this study, we demonstrate that photodynamic nanoparticle-embedded surfaces can swiftly inactivate both enveloped and non-enveloped viruses. We prepared core–shell structured methylene blue (MB)-loaded SiO2 nanoparticles with a high reactive oxygen species (ROS) yield (0.47 ± 0.02). The superior ROS production was maintained after modifying these nanoparticles onto air filter fibers, likely due to the prevention of aggregation-caused quenching effects. Three viruses, including both enveloped and non-enveloped types, were rapidly inactivated within just 12 min (>6 log units) under medium light intensity (660 nm, 30 mW/cm2). Mechanistic studies revealed that envelope glycoproteins are the primary targets for this rapid inactivation. Thus, photodynamic nanoparticle-embedded surfaces offer a straightforward and adaptable strategy in the fight against viral epidemics.

Unveiling the inactivation mechanisms of different viruses in sludge anaerobic digestion based on factors identification and damage analysis

Despite anaerobic digestion having potential for pathogen reduction in sewage sludge, the behaviors of viruses as the primary health concern are rarely studied. This study investigated the inactivation kinetics and mechanisms of four typical virus surrogates with different structures in mesophilic (MAD) and thermophilic (TAD) anaerobic digestion of sludge. Virus inactivation in MAD was virus-type-dependent correspondingly to different function loss. Temperature drove the faster inactivation proceeding for enveloped Phi6, while temperature and ammonia were the critical inactivation factors for nonenveloped MS2, causing genome degradation and protein functional damage. Interaction with sludge solids played critical role in DNA viruses T4 and Phix174 inactivation via inducing host binding function damage. By comparison, TAD enhanced viral protein denaturation, bringing efficient inactivation with reducing heterogeneity among nonenveloped viruses. These insights into unique virus behaviors in anaerobic digestion systems can provide guidance for developing more effective disinfection protocols and improving sludge biosafety.

Mechanisms of AAV2 neutralization by human alpha-defensins.

AAVs are commonly used as gene therapy vectors due to their broad tropism and lack of disease association; however, host innate immune factors, such as human alpha-defensin antimicrobial peptides, can hinder gene delivery. Although it is becoming increasingly evident that human alpha-defensins can block infection by a wide range of nonenveloped viruses, including AAVs, their mechanism of action remains poorly understood. In this study, we describe for the first time how two types of abundant human alpha-defensins neutralize a specific AAV serotype, AAV2. We found that one defensin prevents AAV2 from binding to cells, the first step in infection, while both defensins block a critical later step in AAV2 entry. Our findings support the emerging idea that defensins use a common strategy to block infection by DNA viruses that replicate in the nucleus. Through understanding how innate immune effectors interact with and impede AAV infection, vectors can be developed to bypass these interventions and allow more efficient gene delivery.

Orally bioavailable RORγ/DHODH dual host-targeting small molecules with broad-spectrum antiviral activity

Host-directed antivirals (HDAs) represent an attractive treatment option and a strategy for pandemic preparedness, especially due to their potential broad-spectrum antiviral activity and high barrier to resistance development. Particularly, dual-targeting HDAs offer a promising approach for antiviral therapy by simultaneously disrupting multiple pathways essential for viral replication.Izumerogant (IMU-935) targets two host proteins, (i) the retinoic acid receptor-related orphan receptor γ isoform 1 (RORγ1), which modulates cellular cholesterol metabolism, and (ii) the enzyme dihydroorotate dehydrogenase (DHODH), which is involved in de novo pyrimidine synthesis. Here, we synthesized optimized derivatives of izumerogant and characterized their antiviral activity in comparison to a recently described structurally distinct RORγ/DHODH dual inhibitor. Cell culture-based infection models for enveloped and non-enveloped DNA and RNA viruses, as well as a retrovirus, demonstrated high potency and broad-spectrum activity against human viral pathogens for RORγ/DHODH dual inhibitors at nanomolar concentrations. Comparative analyses with equipotent single-target inhibitors in metabolite supplementation approaches revealed that the dual-targeting mode represents the mechanistic basis for the potent antiviral activity. For SARS-CoV-2, an optimized dual inhibitor completely blocked viral replication in human airway epithelial cells at 5 nM and displayed a synergistic drug interaction with the nucleoside analog molnupiravir. In a SARS-CoV-2 mouse model, treatment with a dual inhibitor alone, or in combination with molnupiravir, reduced the viral load by 7- and 58-fold, respectively.Considering the clinical safety, oral bioavailability, and tolerability of izumerogant in a recent Phase I study, izumerogant-like drugs represent potent dual-targeting antiviral HDAs with pronounced broad-spectrum activity for further clinical development.

Molecular Mechanism of pH-Induced Protrusion Configuration Switching in Piscine Betanodavirus Implies a Novel Antiviral Strategy.

Many viruses contain surface spikes or protrusions that are essential for virus entry. These surface structures can thereby be targeted by antiviral drugs to treat viral infections. Nervous necrosis virus (NNV), a simple nonenveloped virus in the genus of betanodavirus, infects fish and damages aquaculture worldwide. NNV has 60 conspicuous surface protrusions, each comprising three protrusion domains (P-domain) of its capsid protein. NNV uses protrusions to bind to common receptors of sialic acids on the host cell surface to initiate its entry via the endocytic pathway. However, structural alterations of NNV in response to acidic conditions encountered during this pathway remain unknown, while detailed interactions of protrusions with receptors are unclear. Here, we used cryo-EM to discover that Grouper NNV protrusions undergo low-pH-induced compaction and resting. NMR and molecular dynamics (MD) simulations were employed to probe the atomic details. A solution structure of the P-domain at pH 7.0 revealed a long flexible loop (amino acids 311-330) and a pocket outlined by this loop. Molecular docking analysis showed that the N-terminal moiety of sialic acid inserted into this pocket to interact with conserved residues inside. MD simulations demonstrated that part of this loop converted to a β-strand under acidic conditions, allowing for P-domain trimerization and compaction. Additionally, a low-pH-favored conformation is attained for the linker connecting the P-domain to the NNV shell, conferring resting protrusions. Our findings uncover novel pH-dependent conformational switching mechanisms underlying NNV protrusion dynamics potentially utilized for facilitating NNV entry, providing new structural insights into complex NNV-host interactions with the identification of putative druggable hotspots on the protrusion.

Cell Extracts Derived from Cypress and Cedar Show Antiviral Activity against Enveloped Viruses.

The antiviral efficacy of cell-extracts (CEs) derived from cypress (Chamaecyparis obtusa (Siebold & Zucc.) Endl., C. obtusa) and cedar (Cryptomeria japonica (Thunb. ex. L.) D.Don, C. japonica) was assessed using phi6 and MS2 bacteriophages, which are widely accepted surrogate models for enveloped and non-enveloped viruses, in order to verify their potential use as antiviral agents. Our results indicate that CEs derived from C. obtusa are dominantly composed of terpinen-4-ol (18.0%), α-terpinyl acetate (10.1%), bornyl acetate (9.7%), limonene (7.1%), and γ-terpinene (6.7%), while CEs derived from C. japonica are dominantly composed of terpinen-4-ol (48.0%) and α-pinene (15.9%), which exhibited robust antiviral activity against phi6 bacteriophage. Both CEs successfully inactivated the phi6 bacteriophage below the detection limit (10 PFU/mL) within a short exposure time of 30 s (log reduction value, LRV > 4). Through exposure experiments utilizing CEs with content ratios prepared via 2-fold serial dilutions (ranging from 3.13% to 100%), we demonstrated that the antiviral effect could be sustained up to a concentration of 25% (C. obtusa LRV = 3.8, C. japonica LRV > 4.3 at a 25% CE content ratio for each species). However, CEs with content ratios below 12.5% did not produce a significant reduction in bacteriophage concentration and consequently lost their antiviral effects. Conversely, both CEs did not exhibit antiviral activity against MS2 bacteriophage, a non-enveloped virus. Our findings reveal for the first time the potential of CEs derived from C. obtusa and C. japonica for use as antiviral agents specifically targeting enveloped viruses.

Genome microsatellite signature analysis in Adenoviridae family of viruses

Simple sequence repeats (SSRs) are present across both coding & non-coding regions of the genomes of all organisms with diversity in incidence, complexity, and repetition. The present study is focused on Adenoviridae which are non-enveloped viruses consisting of linear double-stranded DNA genome. The members of Adenoviridae are known to cause respiratory illnesses like the common cold (cough, runny nose, mild fever), pneumonia (occasionally), keratoconjunctivitis (infection in the eye known as pink eye), croup and cystitis (inflammation of the bladder). Our investigation aims to extract and analyse SSRs from 68 Adenoviridae genomes. Virus genome sequences were retrieved from NCBI and SSRs extracted from MISA. ETE3 and iTOL were used for the phylogenetic tree construction, annotation and visualization. The genome length of Adenoviridae genomes ranged from 26163 bp to 45667 bp while GC content varied from 33.6 % to 66.9 %. Genome wide analysis revealed a total incidence of 9861 SSRs and 793 cSSRs. The minimum and maximum range of SSR incidence is 112 (A67) to 203 (A61) respectively. The most prevalent mono, di and tri-SSR motif is “A”, “GC/CG” and “GAG/CTC” comprised of 1177, 1859 and 201 occurrences respectively. About 78 % SSRs are present in the coding region in the studied genomes. In terms of protein specific distribution, DNA polymerase enzyme had the highest incidence of 485 SSRs. The presence of mono-SSRs in the A/T region is a marker for host determination and divergence. The average mono-SSRs present in A/T region is 67.14 % and it ranged from 16.22 % to 99.11 %. The high prevalence of mono-SSRs in the A/T region was associated with human and related species as hosts. Further, the clustering of viruses as per their hosts was observed in the phylogenetic tree suggesting the role of host in viral evolution. The presence of unique and conserved cSSRs as genome markers has also been highlighted.

A SARS-CoV-2 peptide antigen purified from bacteria and displayed in a high-density repetitive manner on a virus-like particle could generate anti-SARS-CoV-2 neutralizing antibodies unlike free peptide

SARS-CoV-2 is an enveloped virus. Proteins of the lipid based envelope are not rigidly arranged as compared to non-enveloped viruses lacking lipid based outer layer. Rigidly arranged multivalent display has been reported to be important for potent immune stimulation. We assembled himeric virus-like-particle (VLP) where the core of the particle is composed of the coat protein of Acinetobacter phage. Peptide-based antigen from receptor binding domain (RBD) of SARS-CoV-2 spike protein has been displayed on the coat based VLP matrix using spy-tag/spy-catcher split inteine conjugation system that allows spontaneous, irreversible isopeptide bond formation. Both the matrix as well as peptide have been purified from bacteria which is an easy platform for protein purification. We used the closed state of spike trimer for epitope mapping as this is the conformation of the spike that the immune system visualizes unlike the open state that appears when the virus tries to attach to receptor. Mice based immunization studies were used to test immunogenicity of the antigens. The work demonstrates that peptide-based antigens displayed in high densities can induce neutralizing antibody production unlike free peptide. Careful choice of peptides can deliver better candidates. Also, small size allows easy improvisation. Production in bacteria offers cheaper and robust purification option.

Knowledge, Beliefs and Vaccination Status of Nursing Students about Human Papilloma Virus

Objective: Human papilloma virus (HPV-Human Papilloma Virus) is a small, non-enveloped double-stranded DNA virus from the Papilo mavirada family. This study was conducted to determine the effect of HPV on the knowledge, beliefs and vaccination status of first and fourth year nursing students. Method: Data were collected by personal information form, health belief model scale about human papillomavirus infection and vaccination, and human papillomavirus knowledge scale. Results: The mean perceived benefit score of the students participating in the study was 2.4±0.7 in the first grade and 3.1±0.7 in the fourth grade. The mean perceived sensitivity score was 2.3 ± 0.6 in the first grade and 2.8±0.7 in the fourth grade. The mean perceived severity score was 2.5±0.6 in the first grade and 2.8±0.7 in the fourth grade. The mean perceived barrier score was equal and 2.1±0.4 in all groups. The HPV knowledge scale scores of the students participating in the study consisted of 4 sub-dimensions. The mean score of the general HPV knowledge scale was 4.79±5.6 in the first grade and 16.71±7.9 in the fourth grade. The mean score of HPV screening tests was 3.3±3.7 in the first grade and 9.9±4.0 in the fourth grade. The mean HPV screening test score was 0.4±0.8 in the first grade and 2±1.7 in the fourth grade. The mean score for general HPV vaccination knowledge was 0.7±1.3 in the first grade and 2.8±1.7 in the fourth grade. The mean score for knowledge of HPV vaccination programs was 0.2±0.6 in the first grade and 1.8±1.7 in the fourth grade. Conclusion: In the fight against cervical cancers, which is an important public health problem, the level of HPV knowledge should be high in order for the nursing profession to fulfil all of its education, counselling, guidance, protection and immunisation functions.

Cholesterol-Dependent Membrane Deformation by Metastable Viral Capsids Facilitates Entry.

Nonenveloped viruses employ unique entry mechanisms to breach and infect host cells. Understanding these mechanisms is crucial for developing antiviral strategies. Prevailing perspective suggests that nonenveloped viruses release membrane pore-forming peptides to breach host membranes. However, the precise involvement of the viral capsid in this entry remains elusive. Our study presents direct observations elucidating the dynamically distinctive steps through which metastable reovirus capsids disrupt host lipid membranes as they uncoat into partially hydrophobic intermediate particles. Using both live cells and model membrane systems, our key finding is that reovirus capsids actively deform and permeabilize lipid membranes in a cholesterol-dependent process. Unlike membrane pore-forming peptides, these metastable viral capsids induce more extensive membrane perturbations, including budding, bridging between adjacent membranes, and complete rupture. Notably, cholesterol enhances subviral particle adsorption, resulting in the formation of pores equivalent to the capsid size. This cholesterol dependence is attributed to the lipid condensing effect, particularly prominent at an intermediate cholesterol level. Furthermore, our results reveal a positive correlation between membrane disruption extent and efficiency of viral variants in establishing infection. This study unveils the crucial role of capsid-lipid interaction in nonenveloped virus entry, providing new insights into how cholesterol homeostasis influences virus infection dynamics.

A self-amplifying RNA vaccine prevents enterovirus D68 infection and disease in preclinical models.

The recent emergence and rapid response to severe acute respiratory syndrome coronavirus 2 was enabled by prototype pathogen and vaccine platform approaches, driven by the preemptive application of RNA vaccine technology to the related Middle East respiratory syndrome coronavirus. Recently, the National Institutes of Allergy and Infectious Diseases identified nine virus families of concern, eight enveloped virus families and one nonenveloped virus family, for which vaccine generation is a priority. Although RNA vaccines have been described for a variety of enveloped viruses, a roadmap for their use against nonenveloped viruses is lacking. Enterovirus D68 was recently designated a prototype pathogen within the family Picornaviridae of nonenveloped viruses because of its rapid evolution and respiratory route of transmission, coupled with a lack of diverse anti-enterovirus vaccine approaches in development. Here, we describe a proof-of-concept approach using a clinical stage RNA vaccine platform that induced robust enterovirus D68-neutralizing antibody responses in mice and nonhuman primates and prevented upper and lower respiratory tract infections and neurological disease in mice. In addition, we used our platform to rapidly characterize the antigenic diversity within the six genotypes of enterovirus D68, providing the necessary data to inform multivalent vaccine compositions that can elicit optimal breadth of neutralizing responses. These results demonstrate that RNA vaccines can be used as tools in our pandemic-preparedness toolbox for nonenveloped viruses.

Isothermal Technologies for HPV Detection: Current Trends and Future Perspectives.

The human papillomavirus (HPV) is a non-enveloped DNA virus transmitted through skin-to-skin contact that infects epithelial and mucosal tissue. It has over 200 known genotypes, classified by their pathogenicity as high-risk and low-risk categories. High-risk HPV genotypes are associated with the development of different types of cancers, including cervical cancer, which is a leading cause of mortality in women. In clinical practice and the market, the principal tests used to detect HPV are based on cytology, hybrid detection, and qPCR. However, these methodologies may not be ideal for the required timely diagnosis. Tests have been developed based on isothermal nucleic acid amplification tests (INAATs) as alternatives. These tests offer multiple advantages over the qPCR, such as not requiring specialized laboratories, highly trained personnel, or expensive equipment like thermocyclers. This review analyzes the different INAATs applied for the detection of HPV, considering the specific characteristics of each test, including the HPV genotypes, gene target, the limit of detection (LOD), detection methods, and detection time. Additionally, we discuss the tests available on the market that are approved by the Food and Drug Administration (FDA). Finally, we address the challenges and potential solutions for the large-scale implementation of INAATs, particularly in rural or underserved areas.

Unveiling the Broad-Spectrum Virucidal Potential of Purified Clinoptilolite-Tuff.

Due to its remarkable surface properties, natural clinoptilolite-tuff interacts with a variety of biochemical, pharmaceutical, chemical, and microbiological entities, including human viruses. In the present work, the virucidal activity of purified clinoptilolite-tuff (PCT) was investigated using a variety of viruses, differing in their structure and composition. Influenza A virus, Herpes Simplex virus, Rhinovirus, and Parvovirus were chosen to represent enveloped and non-enveloped viruses with RNA and DNA genomes. Beside human viruses, Canine Parvovirus and bacteriophages T4 and MS2 were used to represent animal and bacterial viruses, respectively. The virucidal activity of PCT was quantified by examining the residual viral activity on susceptible cell lines upon incubation with PCT. A wide range of antiviral efficiencies was observed, ranging from up to 99% for Herpes Simplex virus to no activity for Rhinovirus and both bacteriophages. This study reveals that the virucidal potential of PCT is not universal and depends on a complex set of factors including virus structure and medium composition. The environmental and medical implications of this research are discussed for uses such as wastewater treatment or wound healing.

Ensuring Transfusion Safety: Screening Blood Donors for Human Parvovirus B19.

Ensuring the safety of blood and blood products is a vital aspect of healthcare. The potential for transmission of pathogens through blood and blood products makes transfusion safety a significant concern. Despite advancements in testing methodologies, donated blood products still pose a risk for infection transmission. Human parvovirus B19 (B19V) is a small, single-stranded, non-enveloped DNA virus transmissible parenterally by blood transfusion. B19V causes a wide range of clinical manifestations, which is generally harmless in healthy individuals. B19V infection may cause severe complications, such as aplastic crises, as it affects erythrocyte progenitor cells in individuals with increased erythrocyte turnover. Additionally, B19V can be transmitted from pregnant women to their foetus, potentially causing hydrops fetalis and foetal death. The potential for transmission through blood and blood products makes B19V a significant concern for transfusion safety. In response to the growing recognition of B19V's impact on transfusion safety, various international health organisations have introduced guidelines to minimise its transmission through blood and plasma products. However, the implementation of these guidelines varies globally, with some regions, such as India, still lacking formal protocols for B19V monitoring. This review article explores the existing methodologies for screening blood donors for B19V, assesses the associated transfusion risks, and considers the implications for public health and clinical practice. By emphasising advancements in diagnostic techniques and the challenges of their implementation, this article provides a comprehensive overview of efforts to reduce the transmission of B19V through blood transfusions, thereby ensuring safer blood supplies and improved patient outcomes.