Dengue Virus NS1 Research Articles

Peptides targeting dengue viral nonstructural protein 1 inhibit dengue virus production

Viruses manipulate the life cycle in host cells via the use of viral properties and host machineries. Development of antiviral peptides against dengue virus (DENV) infection has previously been concentrated on blocking the actions of viral structural proteins and enzymes in virus entry and viral RNA processing in host cells. In this study, we proposed DENV NS1, which is a multifunctional non-structural protein indispensable for virus production, as a new target for inhibition of DENV infection by specific peptides. We performed biopanning assays using a phage-displayed peptide library and identified 11 different sequences of 12-mer peptides binding to DENV NS1. In silico analyses of peptide-protein interactions revealed 4 peptides most likely to bind to DENV NS1 at specific positions and their association was analysed by surface plasmon resonance. Treatment of Huh7 cells with these 4 peptides conjugated with N-terminal fluorescent tag and C-terminal cell penetrating tag at varying time-of-addition post-DENV infection could inhibit the production of DENV-2 in a time- and dose-dependent manner. The inhibitory effects of the peptides were also observed in other virus serotypes (DENV-1 and DENV-4), but not in DENV-3. These findings indicate the potential application of peptides targeting DENV NS1 as antiviral agents against DENV infection.

Genetic characterization of dengue virus serotype 2 isolated from dengue fever outbreaks in eastern Uttar Pradesh and western Bihar, India.

Dengue (DEN) is the most common cause of mosquito-borne endemicviral diseases in the tropical and subtropical countries. DEN outbreaks associated with multiple dengue virus (DV) serotypes have been regularly reported in different parts of India. This study was done during DEN outbreaks in 2015 to 2016 in UP and Bihar where DEN-2 was found as the only prevalent serotype. DV-2 was the only serotype amplified in serotype-specific reverse-transcription polymerase chain reaction from sera of 210 (65.21%) out of 322 DV NS1 antigen-positive patients. Further genetic analysis based on full-length envelope (E) protein sequence derived from patient's sera as well as DV isolate showed the circulation of lineages I and III of DV-2 cosmopolitan genotype during 2015 and lineage II during 2016. Finally, the phylogenetic analysis using the E gene sequence revealed that these DV-2 strains have a close genetic relationship with the recently reported DV-2 genotypes from DEN outbreaks reported from different parts of north India. These results showed the circulation of cosmopolitan genotype of DV-2 in eastern Uttar Pradesh and western Bihar, India. The genetic database generated on circulating DV strains in this study will be useful as reference for disease surveillance and strengthening laboratory diagnosis protocols.

Molecular phylogeny and distribution of dengue virus serotypes circulating in Nepal in 2017.

Dengue virus (DENV) infection is endemic in Nepal. Although infection rates are reported annually, little information is available about the circulating viral serotypes and genotypes. Here, we report the results of a multicentre cross-sectional study of DENV serotypes and genotypes sampled from individuals with suspected DENV infection in Nepal in 2017. Of the 50 patients sampled, 40 were serologically positive for DENV NS1, 29 for anti-DENV IgM, 21 for anti-DENV IgG and 14 were positive by qRT-PCR. The three serotypes DENV-1, 2 and 3 were detected and there was no DENV-4. Positive samples from serotyping were subjected to PCR amplification by envelope (E) gene specific primer and subsequent bidirectional sequencing of 5 samples. A time to most recent common ancestor phylogenetic tree was constructed from the new sequences obtained here together with historical DENV-1 and DENV-2 E gene sequences. The DENV-1 isolates (n = 2) from Nepalese individuals were closely related to Indian genotype V, whereas DENV-2 isolates (n = 3) belonged to Cosmopolitan genotype IVa, which is closely related to Indonesian isolates. Historical DENV isolates obtained between 2004 and 2013 clustered with Cosmopolitan IVb, Cosmopolitan IVa, and Asian II genotypes. All Nepalese isolates had different lineages with distinct ancestries. With the exception of isolates obtained in 2004, all other previously published isolates had ancestry to geographically distant part of the world. Molecular analysis revealed dengue epidemics to be comprised of different genotypes of serotype 1 and 2 raising concerns on potential role of different genotypes causing Dengue hemorrhagic fever. Also, our result indicated spread of DENV-2 in non-endemic area such as hilly region of Nepal which was considered to be free of dengue due to high altitude and cold weather.

Molecular Diversity of Dengue Virus Serotypes 1–4 during an Outbreak of Acute Dengue Virus Infection in Theni, India

AbstractBackground: Dengue fever (DF) is caused by an arthropod-borne dengue virus (DENV), has four serotypes and several genotypes. Although having clinical and epidemiological significance, the information on the circulating serotypes/genotypes is scarce in India. Materials and Methods: Blood specimens were collected from the patients suspected of DF and they are tested for DENV NS1 antigen and DENV IgM by ELISA. Antigen-positive samples were further serotyped by reverse transcriptase polymerase chain reaction. Representative samples from each serotype were sequenced to identify the genotypes. Results: All the four DENV serotypes were detected with the pre-dominance of DENV-1 (n = 49; 41.9%). Cases with multiple DENV serotype infections were also identified. Genotyping showed that DENV-1 belonging to genotype I, DENV-2 cosmopolitan (IV), DENV-3 genotype III and DENV-4 genotype I were active in the circulation during the outbreak in 2017. Conclusion: Our study documents the molecular characteristics of DENV circulating in our geographical locality. The detection of heterologous DENV serotypes highlights the importance of regular molecular monitoring for the early recognition of any switch in pre-dominant serotype.

Structural Non-Structural Protein-1 Virus Dengue of Risk Factors on Functional Interference Hepar

The causes of hepatic damage to DHF are mixed. Dengue virus can infect the liver and cause damage.Histopathology of the liver in cases of fatal dengue shows that hepatocytes and Kupffer cells can be thetarget of viral replication and there is involvement of apoptotic mechanism. This study aims to determinethe association between dengue virus NS1 protein and degree of clinical manifestation of dengue infectionwith hepatic dysfunction. Blood samples after taking the serum, were examined for dengue virus NS1protein and SGOT and SGPT examinations at the Surabaya Health Laboratory Center. Data analysis usedChi Square, Fisher's Exact, Anava and Dunnet (a = 0.05). The results shows that the correlated variablesignificantly with dengue hemorrhagic manifestation degree was dengue virus NS protein1 (p = 0.047),with correlation strength of 26.8%. Variables significantly correlated with liver dysfunction were degreeof clinical manifestation of dengue fever (p = 0.037) with correlation strength of 33.5% and dengue virusNS protein1 (p = 0.023) with correlation strength of 29.3%. There was a significant relationship betweenthe presence of dengue virus NS1 protein and the incidence of DHF. There was a significant relationshipbetween the presence of dengue virus NS1 protein with hepatic dysfunction (SGOT and SGPT) in denguevirus infection.

Verification of dengue fever cases imported to the territory of the south Far East.

On biomaterial from 70 people, verification of imported cases of Dengue fever into the south of the Far East from 2012 to 2019 is shown. Serological and virological methods were used, as well as PCR. Using the immunochromatographic rapid test, the Dengue virus (DENV) NS1 antigen and antibodies to DENV (IgM and IgG) were detected in human blood. We examined 12 patients from the infectious diseases department with unknown fever and the blood of 58 people who applied to clinics in Vladivostok after returning from tourist trips. Dengue fever was diagnosed in 23 patients (32.8%), of which antigen was detected in 56%, IgM antibodies in 91.3% and IgG in 52.1%. In 2 cases (8.7%), only antigen was detected in patients. Three strains of the pathogen were isolated by virological methods from 18 blood samples, two of which turned out to be the DENV of the 1st genotype and one - of the DENV of the 2nd genotype. Using RT-PCR, 38 blood samples were tested positive in the immunochromatographic rapid test, of which in 16 cases (42.1%) a DENV marker was detected, in 11 cases it was genotype 1, in three cases genotype 2, and one each - genotypes 3 and 4. 1. The most reliable method of rapid verification (in 100%) the primary infection DENV was the comprehensive determination of antigen and antibodies of the IgM class; 2. With antigenemia, blood should be used to isolate the virus, as well as to diagnose the disease by PCR and to establish the genotype of the DENV; 3. When using only PCR to indicate Dengue virus, a significant proportion of the disease cases will not be diagnosed.

Discovery of Natural Product Compounds as Dengue Virus NS5 Methyltransferase Inhibitor Candidate through in Silico Method

Dengue is a global health problem which predominantly affected the tropical and subtropical region of Asia, Africa, and America. However, there are no available antiviral agents to treat dengue virus (DENV) infection. This study was conducted to utilize natural product compounds as an inhibitor of NS5 Methyltransferase, a viral protein which plays an essential role in the synthesis of DENV RNA. The natural product compounds were collected from the Universal Natural Product Database (UNPD), totaling 229,000 compounds. The in silico screening of the natural product compounds was performed by molecular docking simulation method, which simulates the interaction of the compounds on the active site of the NS5 methyltransferase. From the molecular docking simulation, about 51 compounds showed better affinity and interaction compared to the standard compound, S-Adenosyl-L-Homocysteine (SAH). Then, a series of pharmacological tests were performed to find the best drug candidates by employing DataWarrior and SwissADME software. Finally, three natural product compounds, namely UNPD98966, UNPD183023, and UNDP104952, were regarded as the best inhibitor against NS5 methyltransferase based on its molecular affinity and interaction. These compounds also showed potential as drug candidates due to their desirable pharmacological properties.

Structural Non-Structural Protein-1 Virus Dengue of Risk Factors on Functional Interference Hepar

The causes of hepatic damage to DHF are mixed. Dengue virus can infect the liver and cause damage.Histopathology of the liver in cases of fatal dengue shows that hepatocytes and Kupffer cells can be thetarget of viral replication and there is involvement of apoptotic mechanism. This study aims to determinethe association between dengue virus NS1 protein and degree of clinical manifestation of dengue infectionwith hepatic dysfunction. Blood samples after taking the serum, were examined for dengue virus NS1protein and SGOT and SGPT examinations at the Surabaya Health Laboratory Center. Data analysis usedChi Square, Fisher's Exact, Anava and Dunnet (a = 0.05). The results shows that the correlated variablesignificantly with dengue hemorrhagic manifestation degree was dengue virus NS protein1 (p = 0.047),with correlation strength of 26.8%. Variables significantly correlated with liver dysfunction were degreeof clinical manifestation of dengue fever (p = 0.037) with correlation strength of 33.5% and dengue virusNS protein1 (p = 0.023) with correlation strength of 29.3%. There was a significant relationship betweenthe presence of dengue virus NS1 protein and the incidence of DHF. There was a significant relationshipbetween the presence of dengue virus NS1 protein with hepatic dysfunction (SGOT and SGPT) in denguevirus infection.

New Binding Site Conformations of the Dengue virus NS3 Protease Accessed by long-timescale Molecular Dynamics Simulations

New Binding Site Conformations of the Dengue virus NS3 Protease Accessed by long-timescale Molecular Dynamics Simulations

Implementation of a Multiplex rRT-PCR for Zika, Chikungunya, and Dengue Viruses: Improving Arboviral Detection in an Endemic Region.

Arboviral diagnosis has been complicated throughout the tropical and subtropical Americas by the recent co-circulation of Zika virus (ZIKV), chikungunya virus (CHIKV), and dengue virus (DENV). The aim of this study was to implement a multiplex real-time RT-PCR (rRT-PCR) for ZIKV, CHIKV, and DENV in Paraguay to test patients who were clinically suspected of having dengue. We tested 110 sera from patients who presented to the Hospital de Clínicas in 2016 and had testing for DENV nonstructural protein 1 (NS1; 40 positive and 70 negative). Using a composite reference standard, we confirmed 51 dengue cases (46.4%): 38/40 NS1 positive and 13/70 NS1 negative. Chikungunya virus and ZIKV were detected in one sample each, both were DENV NS1 negative. The NS1 test demonstrated good agreement with rRT-PCR for DENV. However, multiplex rRT-PCR identified a subset of dengue cases and additional arboviral infections that would not be detected if NS1 assays are relied upon for diagnosis.

Dengue virus non-structural protein 1 activates the p38 MAPK pathway to decrease barrier integrity in primary human endothelial cells.

Dengue virus (DENV) causes an estimated 390 million infections worldwide annually, with severe forms of disease marked by vascular leakage. Endothelial cells (EC) are directly responsible for vascular homeostasis and are highly responsive to circulating mediators but are not commonly infected. DENV encodes seven non-structural (NS) proteins; with only one of those, NS1, secreted from infected cells and accumulating in the blood of patients. NS1 has been implicated in the pathogenesis of vascular permeability, but the mechanism is not completely understood. Here we used primary endothelial cells and an array of in vitro approaches to study the effect of NS1 in disease-relevant human ECs. Confocal microscopy demonstrated rapid NS1 internalization by ECs into endosomes with accumulation over time. Transcriptomic and pathway analysis showed significant changes in functions associated with EC homeostasis and vascular permeability. Functional significance of this activation was assessed by trans-endothelial electrical resistance and showed that NS1 induced rapid and transient loss in EC barrier function within 3 h post-treatment. To understand the molecular mechanism by which NS1 induced EC activation, we evaluated the stress-sensing p38 MAPK pathway known to be directly involved in EC permeability and inflammation. WB analysis of NS1-stimulated ECs showed clear activation of p38 MAPK and downstream effectors MAPKAPK-2 and HSP27 with chemical inhibition of the p38 MAP kinase pathway restoring barrier function. Our results suggest that DENV NS1 may be involved in the pathogenesis of severe dengue by activating the p38 MAPK in ECs, promoting increased permeability that characterizes severe disease.

Detection of dengue virus type 2 of Indian origin in acute febrile patients in rural Kenya.

Dengue virus (DENV) has caused recent outbreaks in coastal cities of Kenya, but the epidemiological situation in other areas of Kenya is largely unknown. We investigated the role of DENV infection as a cause of acute febrile disease in non-epidemic settings in rural and urban study areas in Kenya. Altogether, 560 patients were sampled in 2016–2017 in rural Taita–Taveta County (n = 327) and urban slums of Kibera, Nairobi (n = 233). The samples were studied for DENV IgM, IgG, NS1 antigen and flaviviral RNA. IgG seroprevalence was found to be higher in Taita–Taveta (14%) than in Nairobi (3%). Five Taita–Taveta patients were positive for flaviviral RNA, all identified as DENV-2, cosmopolitan genotype. Local transmission in Taita–Taveta was suspected in a patient without travel history. The sequence analysis suggested that DENV-2 strains circulating in coastal and southern Kenya likely arose from a single introduction from India. The molecular clock analyses dated the most recent ancestor to the Kenyan strains a year before the large 2013 outbreak in Mombasa. After this, the virus has been detected in Kilifi in 2014, from our patients in Taita–Taveta in 2016, and in an outbreak in Malindi in 2017. The results highlight that silent transmission occurs between epidemics and also affects rural areas. More information is needed to understand the local epidemiological characteristics and future risks of dengue in Kenya.

A Simple Method for the Design and Development of Flavivirus NS1 Recombinant Proteins Using an In Silico Approach.

Even in countries that are currently not facing a flavivirus epidemic, the spread of mosquito-borne flaviviruses presents an increasing public threat, owing to climate change, international travel, and other factors. Many of these countries lack the resources (viral strains, clinical specimens, etc.) needed for the research that could help cope with the threat imposed by flaviviruses, and therefore, an alternative approach is needed. Using an in silico approach to global databases, we aimed to design and develop flavivirus NS1 recombinant proteins with due consideration towards antigenic variation. NS1 genes analyzed in this study included a total of 6,823 sequences, from Dengue virus (DENV), Japanese encephalitis virus (JEV), West Nile virus (WNV), Zika virus (ZIKV), and Yellow fever virus (YKV). We extracted and analyzed 316 DENV NS1 sequence types (STs), 59 JEV STs, 75 WNV STs, 30 YFV STs, and 43 ZIKV STs using a simple algorithm based on phylogenetic analysis. STs were reclassified according to the variation of the major epitope by MHC II binding. 78 DENV epitope type (EpT), 29 JEV EpTs, 29 WNV EpTs, 12 YFV EpTs, and 5 ZIKV EpTs were extracted according to their major epitopes. Also, frequency results showed that there were dominant EpTs in all flavivirus. Fifteen STs were selected and purified for the expression of recombinant antigen in Escherichia coli by sodium dodecyl sulfate extraction. Our study details a novel in silico approach for the development of flavivirus diagnostics, including a simple way to screen the important peptide regions.

A magnetic immunoconjugate nanoplatform for easy colorimetric detection of the NS1 protein of dengue virus in infected serum.

In this work, as a proof of principle, the design and performance evaluation of a simple, cheap and efficient colorimetric test for the detection of the NS1 protein of dengue virus, assisted by an immunoconjugate of magnetite (Fe3O4) nanoparticles coupled to anti-NS1 antibodies is reported. A monoclonal antibody against the NS1 antigen was covalently immobilized on the surface of superparamagnetic iron oxide nanoparticles (SPIONs ∼ 20 nm) and used for the immunodetection of this protein. When the magnetic immuno-nanoplatform is added into infected serum, it conjugates with the NS1 protein and can then be easily separated using an external magnetic field; then, the recovered immunoconjugate is transferred into a well containing a second immobilized NS1-antibody to form an ELISA-type system. When the NS1 protein is present, a color change to blue is induced by reaction with the Perls reagent, which is consistent with the formation of a SPION-antibody-NS1 antigen-antibody conjugate that confirms infection. No false positives were found when NS1 was not present or a different antibody and the NS1 protein were added into the system. The experimental findings could be extrapolated and scaled up to lead to future developments of simple, quick, and inexpensive, in situ biomolecular diagnostic tests for emergent viral infections.

Serotype-specific differences in the laboratory parameters among hospitalized children with dengue and genetic diversity of dengue viruses circulating in Tamil Nadu, India during 2017.

In 2017, Tamil Nadu, a southern state, had the second highest number of dengue cases from India. In the present study, the serotype-specific differences in the clinical manifestations and laboratory parameters among hospitalized children with dengue were investigated and molecular characterization of the circulating dengue virus (DENV) serotypes during 2017 in Tamil Nadu was performed. Eighty children with dengue-like symptoms consecutively admitted to a tertiary care hospital and positive for DENV NS1 antigen were investigated for DENV serotype utilizing a real-time reverse transcriptase based polymerase chain reaction assay. Complete envelope (E) gene sequencing of the DENV strains was performed. Seventy samples were positive for serotyping (25 DENV-1, 17 DENV-2, six DENV-3, and 22 DENV-4). DENV-4 infections were associated with elevated levels of liver enzymes; Alanine aminotransferase (P = .021) and aspartateaminotransferase (P = .001). However, none of the serotype was associated with any specific clinical features and severe dengue. Asian and American/African genotypes of DENV-1 were cocirculating. The circulating genotype was cosmopolitan for DENV-2 with multiple lineages, genotype III for DENV-3 and genotype I for DENV-4. Unique mutations were present in the 2017 DENV-4 isolates. The present study suggests the association of DENV-4 with elevated liver enzymes in children hospitalized for dengue. Further, the study reports the genetic diversity of DENV circulating in Tamil Nadu during 2017. The study calls for continuous monitoring of the circulating serotypes and genotypes at regional level in India which might result in a region wise database useful in predicting future outbreaks.

Identification of Dengue Fever Markers by Dot Immunoasay

The aim . Development of diagnostic kit for identifying markers of dengue fever at all stages of the disease. Materials and methods . In blood serum from patients with suspected dengue fever, NS1 antigen and specific IgM and IgG were detected by immune chromatography, dot immunoassay, enzyme-linked immunosorbent assay, using commercial test systems, as well as the “Dengue Spectrum” experimental kit. Results and discussion . A diagnostic kit has been developed for the detection of dengue fever markers, based on the mechanism of simultaneous differential detection of the agent NS1 protein and IgM and IgG class antibodies with the formation of specific complexes between markers from the test sample and known capture immune reagents, in a certain order, discretely fixed on a dense substrate. It was found that the effective detection of specific IgG and IgM to dengue virus can be carried out according to the scheme in which IgG is captured on the total antigen of the virus with detection using labeled anti-human IgG antibodies, and IgM is detected by capture on anti-human IgM antibodies with detection using total viral antigen. Detection of dengue virus NS1 protein can be performed using a substrate with immobilized monoclonal antibodies to NS1 and a gold immune sol bound to antibodies against NS1. This protocol of the dot analysis provides the limit for determining the recombinant analogue of the NS1 protein equal to 100 ng/ml. Comparative testing of the kit against the panel of clinical samples showed a good agreement between the results and the data obtained using imported commercial tests. The developed kit can be used for screening clinical samples, both in laboratory and in the field.

Serological tests reveal significant cross-reactive human antibody responses to Zika and Dengue viruses in the Mexican population

Zika virus (ZIKV) is a mosquito-borne flavivirus that has caused recent large outbreaks in the Americas. Given its association with severe congenital defects including microcephaly, distinguishing infections caused by ZIKV from those caused by dengue virus (DENV) is of primordial importance. The objectives of this study were to evaluate the recombinant proteins rEIII-ZIKV (Envelope protein domain III) and rNS1ß-leader-ZIKV (non-structural protein 1) for the serological diagnosis of ZIKV in the Mexican population. We also evaluated potential cross-reactivity in commercial enzyme-linked immunosorbent assays (ELISA) based on the ZIKV NS1 and DENV NS1 proteins. rEIII-ZIKV and rNS1ß-leader-ZIKV proteins were tested with sera from 30 PCR-confirmed ZIKV cases, 50 ZIKV-naive, DENV-exposed subjects with no acute febrile disease, (asymptomatic subjects, AS), and 50 ZIKV-naive and DENV naive AS. Commercial ELISA tests were evaluated with sera from 57 ZIKV and 20 DENV PCR-confirmed cases, and 50 ZIKV-naive, DENV-exposed AS. In-house ELISA assays showed that IgM antibody levels against rEIII-ZIKV and rNS1ß-ZIKV were higher in ZIKV naive, DENV-exposed AS than in acutely infected ZIKV individuals. IgG reactivity was highest for rEIII-ZIKV, and indistinguishable between acutely infected ZIKV cases and DENV exposed AS. Positivity for the Euroimmun Zika IgM assay at 7–10 days was considerably higher in DENV-naive ZIKV patients (86%) than in DENV-exposed ZIKV patients (33%), while 39% of the latter had false-negative anti-ZIKV IgG before 7 days of onset. DENV-exposed ZIKV patients presented lower anti-ZIKV IgM and higher IgG responses similar to a secondary dengue response. Forty-four percent of DENV- exposed acute ZIKV patients were DENV IgM positive with the Panbio Dengue assay, and two (15%) of the DENV-naive ZIKV patients presented false DENV IgG conversion. Given the extensive cross-reactivity to both the NS1 and EDIII proteins in current serological methods, the development of sensitive and specific serological tests to distinguish ZIKV from DENV infections is an urgent priority.

Serine protease inhibitor AEBSF reduces dengue virus infection via decreased cholesterol synthesis

Dengue virus (DENV) infection has evolved into a major global health menace and economic burden due to its intensity and geographic distribution. DENV infection in humans can cause a wide range of symptoms including dengue fever (DF), dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). An antiviral agent that is effective against all four serotypes of DENV is urgently needed to prevent and to manage this condition. Reducing the viral load during the early phase of infection may minimize the chance of patients progressing to more severe DHF or DSS. In this study, we set forth to investigate the anti-viral effect of five commercially available protease inhibitors on DENV infection since both viral and host proteases can contribute to effective viral replication. Previously, the serine protease inhibitor AEBSF [4-(2-aminoethyl) benzene sulfonyl fluoride] has been shown to inhibit DENV NS3 protease activity. The results of the present study revealed that DENV genome replication and protein synthesis were significantly inhibited by AEBSF in a dose-dependent manner. AEBSF inhibited the expression of genes such as 3-hydroxy 3-methyl-glutaryl-CoA synthase (HMGCS), 3-hydroxy-3-methyl-glutaryl-CoA reductase (HMGCR), and low-density lipoprotein receptor (LDLR). Moreover, AEBSF significantly inhibited HMGCR activity and intracellular cholesterol synthesis after DENV infection. The anti-DENV effect of AEBSF was confirmed in all four DENV serotypes and in three different cell lines. These results indicate that AEBSF reduces DENV infection via both viral and host protease activities.

A cross-sectional study of acute dengue infection in paediatric clinics in Cameroon

BackgroundDengue fever is the world’s fastest spreading mosquito borne viral infection. It is prevalent throughout both subtropical and tropical region, and affects over 128 countries. Dengue virus (DENV) infection poses a serious global public health challenge to three billion people, resulting in approximately 200 million cases of morbidity and 50,000 cases of mortality annually. In Cameroon like in most sub-Saharan African countries, DENV infection occur concurrently with other infectious diseases whose symptoms often overlap, rendering differential diagnosis challenging. This study aims at determining the frequency of acute dengue among febrile children under 15 years attending hospitals in some areas of Cameroon.MethodsA total of 961 children under the age of 15 were recruited in a cross-sectional study using systematic sampling technique and by selecting each subject out of the three. The study was conducted in 10 public health centers in Cameroon. Demographic data and risk factors of the subjects were obtained using well-structured questionnaires. Dengue virus NS1 antigen, IgM and IgG were analysed using a Tell me fast® Combo Dengue NS1-IgG/IgM Rapid Test. An in-house ELISA test for dengue specific IgM antibody was equally performed for confirmation. Descriptive statistical analysis was performed using Graph pad version 6.0.ResultsA prevalence of 6.14% acute dengue virus infection was observed among children with febrile illness with a significant difference (p = 0.0488) between males (4.7%) and females (7.7%). In addition, children who reportedly were unprotected from vectors, showed a comparatively higher prevalence of the disease seropositivity than those practicing protective measures.ConclusionDENV infection therefore is an important cause of fever among children in Cameroon. Thus, there is a need to include differential screening for DENV infections as a tool in the management of fever in children in the country.

Host-Specific NS5 Ubiquitination Determines Yellow Fever Virus Tropism.

The recent yellow fever virus (YFV) epidemic in Brazil in 2017 and Zika virus (ZIKV) epidemic in 2015 serve to remind us of the importance of flaviviruses as emerging human pathogens. With the current global flavivirus threat, there is an urgent need for antivirals and vaccines to curb the spread of these viruses. However, the lack of suitable animal models limits the research questions that can be answered. A common trait of all flaviviruses studied thus far is their ability to antagonize interferon (IFN) signaling so as to enhance viral replication and dissemination. Previously, we reported that YFV NS5 requires the presence of type I IFN (IFN-α/β) for its engagement with human signal transducer and activator of transcription 2 (hSTAT2). In this manuscript, we report that like the NS5 proteins of ZIKV and dengue virus (DENV), YFV NS5 protein is able to bind hSTAT2 but not murine STAT2 (mSTAT2). Contrary to what has been demonstrated with ZIKV NS5 and DENV NS5, replacing mSTAT2 with hSTAT2 cannot rescue the YFV NS5-STAT2 interaction, as YFV NS5 is also unable to interact with hSTAT2 in murine cells. We show that the IFN-α/β-dependent ubiquitination of YFV NS5 that is required for STAT2 binding in human cells is absent in murine cells. In addition, we demonstrate that mSTAT2 restricts YFV replication in vivo These data serve as further impetus for the development of an immunocompetent mouse model that can serve as a disease model for multiple flaviviruses.IMPORTANCE Flaviviruses such as yellow fever virus (YFV), Zika virus (ZIKV), and dengue virus (DENV) are important human pathogens. A common flavivirus trait is the antagonism of interferon (IFN) signaling to enhance viral replication and spread. We report that like ZIKV NS5 and DENV NS5, YFV NS5 binds human STAT2 (hSTAT2) but not mouse STAT2 (mSTAT2), a type I IFN (IFN-α/β) pathway component. Additionally, we show that contrary to what has been demonstrated with ZIKV NS5 and DENV NS5, YFV NS5 is unable to interact with hSTAT2 in murine cells. We demonstrate that mSTAT2 restricts YFV replication in mice and that this correlates with a lack of IFN-α/β-induced YFV NS5 ubiquitination in murine cells. The lack of suitable animal models limits flavivirus pathogenesis, vaccine, and drug research. These data serve as further impetus for the development of an immunocompetent mouse model that can serve as a disease model for multiple flaviviruses.