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genetically engineered synthetic miniaturized versions of plasmodium falciparum uvrd helicase are catalytically active.helicases catalyze unwinding of double stranded nucleic acids in an energy-dependent manner. we have reported characterization of uvrd helicase from plasmodium falciparum. we reported that the n-terminal and c-terminal fragments of pfuvrd contain characteristic atpase and dna helicase activities. here we report the generation and characterization of a genetically engineered version of pfuvrd and its derivatives. this synthetic uvrd (sud) contains all the conserved domains of pfuvrd but only the ...201424608129
dengue virus serotype 1 non-structural protein ns5 expression interferes with hiv replication in a cd4+ t-cell line. 201424470566
cpb1 of aedes aegypti interacts with denv2 e protein and regulates intracellular viral accumulation and release from midgut cells.aedes aegypti is a principal vector responsible for the transmission of dengue viruses (denv). to date, vector control remains the key option for dengue disease management. to develop new vector control strategies, a more comprehensive understanding of the biological interactions between denv and ae. aegypti is required. in this study, a cdna library derived from the midgut of female adult ae. aegypti was used in yeast two-hybrid (y2h) screenings against denv2 envelope (e) protein. among the man ...201425521592
identification and profiling of conserved and novel micrornas in laodelphax striatellus in response to rice black-streaked dwarf virus (rbsdv) infection.micrornas (mirnas) are small non-coding endogenous rna molecules that play important roles in various biological processes. this study examined microrna profiles of laodelphax striatellus using the small rna libraries derived from virus free (vf) and rice black-streaked dwarf virus (rbsdv) infected (rb) insects. a total of 59 mature mirnas (46 mirna families) were identified as conserved insect mirnas in both vf and rb libraries. among these conserved mirnas, 24 were derived from the two arms of ...201426484150
phospholipase a2 isolated from the venom of crotalus durissus terrificus inactivates dengue virus and other enveloped viruses by disrupting the viral envelope.the flaviviridae family includes several virus pathogens associated with human diseases worldwide. within this family, dengue virus is the most serious threat to public health, especially in tropical and sub-tropical regions of the world. currently, there are no vaccines or specific antiviral drugs against dengue virus or against most of the viruses of this family. therefore, the development of vaccines and the discovery of therapeutic compounds against the medically most important flaviviruses ...201425383618
investigating the efficacy of monovalent and tetravalent dengue vaccine formulations against denv-4 challenge in ag129 mice.dengue (den) is the most important mosquito-borne viral disease, with a major impact on global health and economics, caused by four serologically and distinct viruses termed denv-1 to denv-4. currently, there is no licensed vaccine to prevent den. we have developed a live attenuated tetravalent denv vaccine candidate (tdv) (formally known as denvax) that has shown promise in preclinical and clinical studies and elicits neutralizing antibody responses to all four denvs. as these responses are low ...201425239488
characterization of a novel insect-specific flavivirus from brazil: potential for inhibition of infection of arthropod cells with medically important flaviviruses.in the past decade, there has been an upsurge in the number of newly described insect-specific flaviviruses isolated pan-globally. we recently described the isolation of a novel flavivirus (tentatively designated 'nhumirim virus'; nhuv) that represents an example of a unique subset of apparently insect-specific viruses that phylogenetically affiliate with dual-host mosquito-borne flaviviruses despite appearing to be limited to replication in mosquito cells. we characterized the in vitro growth p ...201425146007
rna virus reverse genetics and vaccine design.rna viruses are capable of rapid spread and severe or potentially lethal disease in both animals and humans. the development of reverse genetics systems for manipulation and study of rna virus genomes has provided platforms for designing and optimizing viral mutants for vaccine development. here, we review the impact of rna virus reverse genetics systems on past and current efforts to design effective and safe viral therapeutics and vaccines.201424967693
substitution of ns5 n-terminal domain of dengue virus type 2 rna with type 4 domain caused impaired replication and emergence of adaptive mutants with enhanced fitness.flavivirus ns3 and ns5 are required in viral replication and 5'-capping. ns3 has ns2b-dependent protease, rna helicase, and 5'-rna triphosphatase activities. ns5 has 5'-rna methyltransferase (mt)/guanylyltransferase (gt) activities within the n-terminal 270 amino acids and the rna-dependent rna polymerase (pol) activity within amino acids 271-900. a chimeric ns5 containing the d4mt/d4gt and the d2pol domains in the context of wild-type (wt) d2 rna was constructed. rnas synthesized in vitro were ...201424904061
serological evidence of widespread circulation of west nile virus and other flaviviruses in equines of the pantanal, brazil.a recent study reported neutralizing antibodies to west nile virus (wnv) in horses from four ranches of southern pantanal. to extend that study, a serosurvey for wnv and 11 brazilian flaviviruses was conducted with 760 equines, 238 sheep and 61 caimans from 17 local cattle ranches. among the tested equines, 32 were collected from a ranch where a neurologic disorder outbreak had been recently reported. the sera were initially screened by using a blocking elisa and then titrated by 90% plaque-redu ...201424551266
susceptibility of florida aedes aegypti and aedes albopictus to dengue viruses from puerto rico.locally acquired dengue cases in the continental u.s. are rare. however, outbreaks of dengue-1 during 2009, 2010, and 2013 in florida and dengue-1 and -2 in texas suggest vulnerability to transmission. travel and commerce between puerto rico and the u.s. mainland is common, which may pose a risk for traveler-imported dengue cases. mosquitoes were collected in florida and used to evaluate their susceptibility to dengue viruses (denv) from puerto rico. aedes aegypti and ae. albopictus were suscept ...201425424270
mechanism for maturation-related reorganization of flavivirus glycoproteins.flaviviruses, such as dengue, west nile, and yellow fever viruses, assemble as fusion-incompetent particles and subsequently undergo a large reorganization of their glycoprotein envelope resulting in formation of mature infectious virions. here we used a combination of three-dimensional cryo-electron tomography and two-dimensional image analysis to study pleomorphic maturation intermediates of dengue virus 2. icosahedral symmetries of immature and mature regions within one particle were mismatch ...201424252771
vectors expressing chimeric japanese encephalitis dengue 2 viruses.vectors based on self-replicating rnas (replicons) of flaviviruses are becoming powerful tool for expression of heterologous genes in mammalian cells and development of novel antiviral and anticancer vaccines. we constructed two vectors expressing chimeric viruses consisting of attenuated sa14-14-2 strain of japanese encephalitis virus (jev) in which the prm/m-e genes were replaced fully or partially with those of dengue 2 virus (denv-2). these vectors, named pjed2 and pjed2-1770 were transfecte ...201425518715
inhibitory effects of curcumin on dengue virus type 2-infected cells in vitro.curcumin, a traditional chinese and indian treatment for many diseases, has recently been found to alter the in vitro infection processes of various viruses, including hepatitis c virus, human immunodeficiency virus, coxsackievirus, and japanese encephalitis virus. the present study evaluated the cellular effects of curcumin in an in vitro (cellular) infection model of dengue virus. within a dose range of 10 to 30 μm and a treatment period of 24 hours, the cytotoxicity of curcumin was low, as de ...201424081825
generation and characterization of a rat monoclonal antibody against the rna polymerase protein from dengue virus-2.dengue virus (denv) rna replication requires 2 viral proteins, non-structural protein 3 (ns3) and ns5. ns5 consists of 2 functional domains: a methyltransferase (mtase) domain involved in rna cap formation and located in the amino terminal region and a rna-dependent rna polymerase domain essential for virus replication and located in the carboxyl terminal region. to gain additional insight into the structural interactions between viral proteins and cellular factors involved in denv rna replicati ...201424063571
the ph dependence of flavivirus envelope protein structure: insights from molecular dynamics simulations.the flavivirus membrane fusion is triggered by the acid ph of the endosomes after virus endocytosis. the proposed mechanism involves changes in the protonation state of conserved histidine residues of the e protein present in the viral surface that undergoes a series of structural rearrangements that result in the fusion between the endosome and viral bilayers. we studied the ph dependence of e protein rearrangements of dengue virus type 2, used as a model, in the ph range experimented by the vi ...201423964620
coordinated regulation of autophagy and apoptosis determines endothelial cell fate during dengue virus type 2 infection.dengue is the most prevalent mosquito-borne viral disease in tropical regions. severe cases may progress to dengue hemorrhagic fever, suggesting vascular endothelial dysfunction in disease pathogenesis. in our previous study, we found that dengue virus type 2 (denv2) induced apoptosis of vascular endothelial cells via fasl/fas- and xiap-associated factor 1 (xaf1)-dependent pathways. in this paper, we demonstrate that denv2 can induce autophagy in primary human umbilical vein endothelial cells (h ...201425138703
production of a neutralizing antibody against envelope protein of dengue virus type 2 using the linear array epitope technique.dengue virus (denv; genus flavivirus) contains a positive-stranded rna genome. binding of denv to host cells is mediated through domain iii of the viral envelope protein. many therapeutic mabs against domain iii have been generated and characterized because of its high antigenicity. we have previously established a novel pcr method named the linear array epitope (lae) technique for producing monoclone-like polyclonal antibodies. to prove this method could be utilized to produce antibody against ...201424948392
capsid protein: evidences about the partial protective role of neutralizing antibody-independent immunity against dengue in monkeys.the role of cellular immune response in dengue virus infection is not yet fully understood. only few studies in murine models propose that cd8(+) t-cells are associated with protection from infection and disease. at the light of recent reports about the protective role of cd8(+) t-cells in humans and the no correlation between neutralizing antibodies and protection observed in several studies, a vaccine based on cell-mediated immunity constitute an attractive approach. our group has developed a ...201424889226
recombinant dengue 2 virus ns3 protein conserves structural antigenic and immunological properties relevant for dengue vaccine design.the ns3 protein is a multifunctional non-structural protein of flaviviruses implicated in the polyprotein processing. the predominance of cytotoxic t cell lymphocytes epitopes on the ns3 protein suggests a protective role of this protein in limiting virus replication. in this work, we studied the antigenicity and immunogenicity of a recombinant ns3 protein of the dengue virus 2. the full-length ns3 gene was cloned and expressed as a his-tagged fusion protein in escherichia coli. the pns3 protein ...201424854144
a tetravalent dengue vaccine containing a mix of domain iii-p64k and domain iii-capsid proteins induces a protective response in mice.recombinant fusion proteins containing domain iii of the dengue virus envelope protein fused to the p64k protein from neisseria meningitidis and domain iii of dengue virus type 2 (d2) fused to the capsid protein of this serotype were immunogenic and conferred protection in mice against lethal challenge, as reported previously. combining the domain iii-p64k recombinant proteins of dengue virus types 1, 3 and 4 (d1, d3, and d4) with the domain iii-capsid protein from d2, we obtained a novel tetrav ...201424841761
chloroquine interferes with dengue-2 virus replication in u937 cells.the objective of this study was to investigate the use of chloroquine (clq) as an antiviral agent against dengue. chloroquine, an amine acidotropic drug known to affect intracellular exocytic pathways by increasing endosomal ph, was used in the in vitro treatment of u937 cells infected with dengue virus type 2 (denv-2). viral replication was assessed by quantification of virus produced through detection of copy numbers of denv-2 rna, plaque assay and indirect immunofluorescence. qrt-pcr and plaq ...201424773578
interferon-mediated isg15 conjugation restricts dengue virus 2 replication.isgylation, an ubiquitin-like post-translational modification by isg15, has been reported to participate in the interferon (ifn)-mediated antiviral response. in this study, we analyzed the functional role of isgylation in dengue virus 2 (denv-2) replication. overexpression of isg15 was found to significantly suppress the amount of extracellular infectious virus released, while intracellular viral rna was unaffected. this effect was not observed with a conjugation-defective isg15 mutant. in addit ...201424769207
evaluation of dengue antiviral candidates in vivo in mouse model.in vivo evaluation of antiviral compounds can serve as criteria in the drug discovery process for selection of compounds that are suitable to enter late preclinical studies and further development. dengue virus serotypes 1-4 can infect and replicate in the interferon type i and type ii receptor deficient mice (ag129). here we describe the use of a mouse-adapted dengue 2 virus strain (s221) that has been used to develop a robust lethal model of infection. treatment with small molecule inhibitors ...201424696350
construction of plasmid, bacterial expression, purification, and assay of dengue virus type 2 ns5 methyltransferase.dengue virus (denv), a member of mosquito-borne flavivirus, causes self-limiting dengue fever as well as life-threatening dengue hemorrhagic fever and dengue shock syndrome. its positive sense rna genome has a cap at the 5'-end and no poly(a) tail at the 3'-end. the viral rna encodes a single polyprotein, c-prm-e-ns1-ns2a-ns2b-ns3-ns4a-ns4b-ns5. the polyprotein is processed into 3 structural proteins (c, prm, and e) and 7 nonstructural (ns) proteins (ns1, ns2a, ns2b, ns3, ns4a, ns4b, ns5). ns3 a ...201424696348
targeted mutagenesis of dengue virus type 2 replicon rna by yeast in vivo recombination.the use of cdna infectious clones or subgenomic replicons is indispensable in studying flavivirus biology. mutating nucleotides or amino acid residues gives important clues to their function in the viral life cycle. however, a major challenge to the establishment of a reverse genetics system for flaviviruses is the instability of their nucleotide sequences in escherichia coli. thus, direct cloning using conventional restriction enzyme-based procedures usually leads to unwanted rearrangements of ...201424696336
changes in antiviral susceptibility to entry inhibitors and endocytic uptake of dengue-2 virus serially passaged in vero or c6/36 cells.the aim of the present study was to analyze the influence of virus origin, mammalian or mosquito cell-derived, on antiviral susceptibility of denv-2 to entry inhibitors and the association of this effect with any alteration in the mode of entry into the cell. to this end, ten serial passages of denv-2 were performed in mosquito c6/36 cells or monkey vero cells and the antiviral susceptibility of each virus passage to sulfated polysaccharides (sps), like heparin and carrageenans, was evaluated by ...201424583230
development of a novel dna-launched dengue virus type 2 infectious clone assembled in a bacterial artificial chromosome.major progress in dengue virus (denv) biology has resulted from the use of infectious clones obtained through reverse genetics. the construction of these clones is commonly based on high- or low-copy number plasmids, yeast artificial chromosomes, yeast-escherichia coli shuttle vectors, and bacterial artificial chromosomes (bacs). prokaryotic promoters have consistently been used for the transcription of these clones. the goal of this study was to develop a novel denv infectious clone in a bac un ...201424342140
dengue virus subgenomic rna induces apoptosis through the bcl-2-mediated pi3k/akt signaling pathway.we report a rna species of 429 nucleotides derived from the 3' untranslated region of the viral genome in dengue 2 virus (denv-2) infected cells. the 3' terminal of viral rna contained specific conserved structures that are important for the production of subgenomic rna. transient replicon assays suggested that loss of this small rna has little effect on viral replication, and genetic analysis using recombinant viruses demonstrated that the existence of this subgenomic rna is not essential for t ...201424314632
identification of novel thiadiazoloacrylamide analogues as inhibitors of dengue-2 virus ns2b/ns3 protease.dengue virus is endemic throughout tropical and subtropical regions, and cause severe epidemic diseases. the ns2b/ns3 protease is a promising drug target for dengue virus. herein, we report the discovery and modification of a novel class of thiadiazoloacrylamide derivatives with potent inhibitory activity against the ns2b/ns3 protease. thiadiazolopyrimidinone 1 was firstly determined as a new chemical structure against ns2b/ns3 from a commercial compound library. then, we sought to identify simi ...201425438757
limonoids from the seeds of swietenia macrophylla with inhibitory activity against dengue virus 2.fractionation of an ethanol-soluble extract of the seeds of swietenia macrophylla yielded six new limonoids, swielimonoids a-f (1-6), along with 20 known compounds. compounds 1 and 2, mexicanolide-type limonoids, were assigned with an α,β-unsaturated δ-lactone moiety (ring d) and a c═c bond between c-8 and c-30. compounds 3-6 could be categorized as highly oxygenated phragmalin-type limonoids. the structures of these new compounds were elucidated through the interpretation of spectroscopic data. ...201425330401
increased replicative fitness of a dengue virus 2 clade in native mosquitoes: potential contribution to a clade replacement event in nicaragua.the four dengue virus (denv) serotypes (denv serotype 1 [denv-1] to denv-4) are transmitted by aedes aegypti and a. albopictus mosquitoes, causing up to 390 million denv infections worldwide each year. we previously reported a clade replacement of the denv-2 asian-american genotype ni-1 clade by the ni-2b clade in managua, nicaragua. here, we describe our studies of the replicative ability of ni-1 and ni-2b viruses in an a. aegypti cell line (aag2) and a. aegypti mosquitoes reared from eggs coll ...201425187539
safety and immunogenicity of a rederived, live-attenuated dengue virus vaccine in healthy adults living in thailand: a randomized trial.safety and immunogenicity of two formulations of a live-attenuated tetravalent dengue virus (tden) vaccine produced using rederived master seeds from a precursor vaccine were tested against a placebo control in a phase ii, randomized, double blind trial (nct00370682). two doses were administered 6 months apart to 120 healthy, predominantly flavivirus-primed adults (87.5% and 97.5% in the two vaccine groups and 92.5% in the placebo group). symptoms and signs reported after vaccination were mild t ...201424865677
genesv - an approach to help characterize possible variations in genomic and protein sequences.a computational approach for identification and assessment of genomic sequence variability (genesv) is described. for a given nucleotide sequence, genesv collects information about the permissible nucleotide variability (changes that potentially preserve function) observed in corresponding regions in genomic sequences, and combines it with conservation/variability results from protein sequence and structure-based analyses of evaluated protein coding regions. genesv was used to predict effects (f ...201424453480
high-throughput quantitative proteomic analysis of dengue virus type 2 infected a549 cells.disease caused by dengue virus is a global health concern with up to 390 million individuals infected annually worldwide. there are no vaccines or antiviral compounds available to either prevent or treat dengue disease which may be fatal. to increase our understanding of the interaction of dengue virus with the host cell, we analyzed changes in the proteome of human a549 cells in response to dengue virus type 2 infection using stable isotope labelling in cell culture (silac) in combination with ...201424671231
amodiaquine, an antimalarial drug, inhibits dengue virus type 2 replication and infectivity.dengue virus serotypes 1-4 (denv1-4) are transmitted by mosquitoes which cause most frequent arboviral infections in the world resulting in ∼390 million cases with ∼25,000 deaths annually. there is no vaccine or antiviral drug currently available for human use. compounds containing quinoline scaffold were shown to inhibit flavivirus ns2b-ns3 protease (ns2b-ns3pro) with good potencies. in this study, we screened quinoline derivatives, which are known antimalarial drugs for inhibition of denv2 and ...201424680954
synthesis and evaluation of imidazole-4,5- and pyrazine-2,3-dicarboxamides targeting dengue and yellow fever virus.the results of a high-throughput screening assay using the dengue virus-2 replicon showed that the imidazole 4,5-dicarboxamide (i45dc) derivative (15a) has a high dengue virus inhibitory activity. based on 15a as a lead compound, a novel class of both disubstituted i45dcs and the resembling pyrazine 2,3-dicarboxamides (p23dcs) were synthesized. here, we report on their in vitro inhibitory activity against dengue virus (denv) and yellow fever virus (yfv). some of these first generation compounds ...201425285371
thioaptamers targeting dengue virus type-2 envelope protein domain iii.thioaptamers targeting the dengue-2 virus (denv-2) envelope protein domain iii (ediii) were developed. ediii, which contains epitopes for binding neutralizing antibodies, is the putative host-receptor binding domain and is thus an attractive target for development of vaccines, anti-viral therapeutic and diagnostic agents. thioaptamer denta-1 bound to denv-2 ediii adjacent to a known neutralizing antibody binding site with a dissociation constant of 154nm.201425261724
vector competence of aedes aegypti populations from kilifi and nairobi for dengue 2 virus and the influence of temperature.susceptibility of ae. aegypti mosquito to dengue virus (denv) varies geographically and can be influenced by climatic factors such as temperature, which affect the incidence, seasonality and distribution of vector-borne diseases. the first outbreak of dengue fever (df) in kenya occured in 1982 in the coastal towns of malindi and kilifi. unlike nairobi where no active dengue transmission has been reported, df is currently re-emerging at the coast causing major outbreaks. this study investigated t ...201425223760
dengue virus 2 american-asian genotype identified during the 2006/2007 outbreak in piauí, brazil reveals a caribbean route of introduction and dissemination of dengue virus in brazil.dengue virus (denv) is the most widespread arthropod-borne virus, and the number and severity of outbreaks has increased worldwide in recent decades. dengue is caused by denv-1, denv- 2, denv-3 and denv-4 which are genetically distant. the species has been subdivided into genotypes based on phylogenetic studies. denv-2, which was isolated from dengue fever patients during an outbreak in piaui, brazil in 2006/2007 was analyzed by sequencing the envelope (e) gene. the results indicated a high simi ...201425127366
effect of dengue-2 virus infection on protein expression in the salivary glands of aedes aegypti mosquitoes.dengue virus (denv) is the most important mosquito-transmitted flavivirus that is transmitted throughout the tropical and subtropical regions of the world. the primary mosquito vector of denv in urban locations is aedes aegypti. key to understanding the transmission of denv is the relationship between pathogen and vector. accordingly, we report our preliminary characterization of the differentially expressed proteins from ae. aegypti mosquitoes after denv infection. we investigated the virus-vec ...201424445208
fitness impact and stability of a transgene conferring resistance to dengue-2 virus following introgression into a genetically diverse aedes aegypti strain.in 2006, we reported a mariner (mos1)-transformed aedes aegypti line, carb77, which was highly resistant to dengue-2 virus (denv2). carb77 mosquitoes expressed a denv2-specific inverted-repeat (ir) rna in midgut epithelial cells after ingesting an infectious bloodmeal. the ir-rna formed double-stranded denv2-derived rna, initiating an intracellular antiviral rna interference (rnai) response. however, carb77 mosquitoes stopped expressing the ir-rna after 17 generations in culture and lost their d ...201424810399
spatiotemporal dynamics of denv-2 asian-american genotype lineages in the americas.the asian/american (as/am) genotype of dengue virus type 2 (denv-2) has been evolving in the americas over the last 30 years, leading to several waves of dengue epidemics and to the emergence of different viral lineages in the region. in this study, we investigate the spatiotemporal dissemination pattern of the denv-2 lineages at a regional level. we applied phylogenetic and phylogeographic analytical methods to a comprehensive data set of 582 denv-2 e gene sequences of the as/am genotype isolat ...201424897118
infection with dengue-2 virus alters proteins in naturally expectorated saliva of aedes aegypti mosquitoes.dengue virus (denv) is responsible for up to approximately 300 million infections and an increasing number of deaths related to severe manifestations each year in affected countries throughout the tropics. it is critical to understand the drivers of this emergence, including the role of vector-virus interactions. when a denv-infected aedes aegypti mosquito bites a vertebrate, the virus is deposited along with a complex mixture of salivary proteins. however, the influence of a denv infection upon ...201424886023
dengue virus type 2 (denv2)-induced oxidative responses in monocytes from glucose-6-phosphate dehydrogenase (g6pd)-deficient and g6pd normal subjects.dengue virus is endemic in peninsular malaysia. the clinical manifestations vary depending on the incubation period of the virus as well as the immunity of the patients. glucose-6-phosphate dehydrogenase (g6pd) deficiency is prevalent in malaysia where the incidence is 3.2%. it has been noted that some g6pd-deficient individuals suffer from more severe clinical presentation of dengue infection. in this study, we aim to investigate the oxidative responses of denv2-infected monocytes from g6pd-def ...201424625456
development of vhh antibodies against dengue virus type 2 ns1 and comparison with monoclonal antibodies for use in immunological diagnosis.the possibility of using variable domain heavy-chain antibodies (vhh antibodies) as diagnostic tools for dengue virus (denv) type 2 ns1 protein was investigated and compared with the use of conventional monoclonal antibodies. after successful expression of denv type 2 ns1 protein, the genes of vhh antibodies against ns1 protein were biopanned from a non-immune llama library by phage display. vhh antibodies were then expressed and purified from escherichia coli. simultaneously, monoclonal antibod ...201424751715
molecular determinants of dengue virus 2 envelope protein important for virus entry in fcγriia-mediated antibody-dependent enhancement of infection.antibody-dependent enhancement (ade) of infection may cause severe illness in patients suffering a secondary infection by a heterologous dengue virus (denv) serotype. during ade of infection, cross-reactive non- or poorly-neutralizing antibodies form infectious virus-ab complexes with the newly infecting serotype and enhance virus infection by binding to the fcγ receptors (fcγr) on fcγr-bearing cells. in this study, we determined that molecular determinants of denv2 envelope protein critical for ...201424889243
lineage ii of southeast asian/american denv-2 is associated with a severe dengue outbreak in the peruvian amazon.during 2010 and 2011, the loreto region of peru experienced a dengue outbreak of unprecedented magnitude and severity for the region. this outbreak coincided with the reappearance of dengue virus-2 (denv-2) in loreto after almost 8 years. whole-genome sequence indicated that denv-2 from the outbreak belonged to lineage ii of the southeast asian/american genotype and was most closely related to viruses circulating in brazil during 2007 and 2008, whereas denv-2 previously circulating in loreto gro ...201425002298
cd8+ t-cell responses in flavivirus-naive individuals following immunization with a live-attenuated tetravalent dengue vaccine candidate.we are developing a live-attenuated tetravalent dengue vaccine (tdv) candidate based on an attenuated dengue 2 virus (tdv-2) and 3 chimeric viruses containing the premembrane and envelope genes of dengue viruses (denvs) -1, -3, and -4 expressed in the context of the attenuated tdv-2 genome (tdv-1, tdv-3, and tdv-4, respectively). in this study, we analyzed and characterized the cd8(+) t-cell response in flavivirus-naive human volunteers vaccinated with 2 doses of tdv 90 days apart via the subcut ...201525943203
screening analogs of β-og pocket binder as fusion inhibitor of dengue virus 2.dengue is an infectious disease caused by dengue virus (denv) and transmitted between human hosts by mosquitoes. recently, indonesia was listed as a country with the highest cases of dengue by the association of southeast asian nations. the current treatment for dengue disease is supportive therapy; there is no antiviral drug available in the market against dengue. therefore, a research on antiviral drug against dengue is very important, especially to prevent outbreak explosion. in this research ...201526617459
evaluation of simultaneous transmission of chikungunya virus and dengue virus type 2 in infected aedes aegypti and aedes albopictus (diptera: culicidae).the simultaneous transmission of chikungunya virus (chikv) and dengue viruses (denv) has been a major public health concern because of their sympatric distribution and shared mosquito vectors. groups of aedes aegypti (l.) and aedes albopictus (skuse) were orally infected with 1.5 × 10(5) pfu/ml of chikv and 3.2 × 10(6) ffu/ml of denv-2 simultaneously or separately in inverse orders and evaluated for dissemination and transmission by qrt-pcr. simultaneous dissemination of both viruses was detecte ...201526334820
dataset on protein composition of a human plasma sub-proteome able to modulate the dengue 2 virus infection in huh 7.5 cells.the four serotypes of dengue virus (denv1-4) are the causal agents of the emerging disease dengue fever and its severe forms. denv is inoculated into human blood through a mosquito bite. thus, plasma is an important media for denv dissemination in infected persons and several important interactions should take place for the virus with human plasma proteins that strongly influence or may determine the course of the infection. this dataset contains 239 proteins identified in the elution fractions ...201526862582
effect of larval competition on extrinsic incubation period and vectorial capacity of aedes albopictus for dengue virus.despite the growing awareness that larval competition can influence adult mosquito life history traits including susceptibility to pathogens, the net effect of larval competition on human risk of exposure to mosquito-borne pathogens remains poorly understood. we examined how intraspecific larval competition affects dengue-2 virus (denv-2) extrinsic incubation period and vectorial capacity of its natural vector aedes albopictus. adult ae. albopictus from low and high-larval density conditions wer ...201525951173
an epitope-substituted dna vaccine improves safety and immunogenicity against dengue virus type 2.dengue virus (denv), a global disease, is divided into four serotypes (denv1-4). cross-reactive and non-neutralizing antibodies against envelope (e) protein of denv bind to the fcγ receptors (fcγr) of cells, and thereby exacerbate viral infection by heterologous serotypes via antibody-dependent enhancement (ade). identification and modification of enhancing epitopes may mitigate enhancement of denv infection. in this study, we characterized the cross-reactive db21-6 and db39-2 monoclonal antibod ...201526135599
antibody avidity following secondary dengue virus type 2 infection across a range of disease severity.the four dengue virus serotypes (denv1-4) are responsible for the most prevalent mosquito-borne viral illness in humans. denv causes a spectrum of disease from self-limiting dengue fever (df) to severe, life-threatening dengue hemorrhagic fever/dengue shock syndrome (dhf/dss). antibodies from one infection can contribute to either protection or increased disease severity in a subsequent infection with a distinct denv serotype. the effectiveness of the antibody response is modulated by both the a ...201526209381
ifi6 inhibits apoptosis via mitochondrial-dependent pathway in dengue virus 2 infected vascular endothelial cells.dengue hemorrhagic fever (dhf)/dengue shock syndrome (dss) is a fatal infectious disease that demands an effective treatment. interferon (ifn)-stimulated genes (isgs) induced by dengue virus (denv) exert antiviral effects. among isgs, ifn-α inducible gene 6 (ifi6) was increased in denv infected human umbilical vascular endothelial cells (huvecs) by microarray analysis in our previous study. however, its function is incompletely understood. in this study, we confirmed that ifi6 was markedly induc ...201526244642
introduction and evolution of dengue virus type 2 in pakistan: a phylogeographic analysis.pattern of dengue periodic epidemics through the years along with sporadic cases of dengue hemorrhagic fever followed by a severe 2011 epidemic of dengue fever in pakistan make pakistan a dengue endemic country. to study the entry and evolution of dengue virus serotype 2 (denv-2) in pakistan, we sequenced three full length genomes and 24 complete envelope sequences of denv-2 from the years 2010, 2011 and 2013 collected from punjab province of pakistan.201526395339
characterization of n-glycan structures on the surface of mature dengue 2 virus derived from insect cells.denv envelope glycoprotein (e) is responsible for interacting with host cell receptors and is the main target for the development of a dengue vaccine based on an induction of neutralizing antibodies. it is well known that denv e glycoprotein has two potential n-linked glycosylation sites at asn67 and asn153. the n-glycans of e glycoprotein have been shown to influence the proper folding of the protein, its cellular localization, its interactions with receptors and its immunogenicity. however, th ...201526208004
complete genome sequence of an atypical dengue virus serotype 2 lineage isolated in brazil.here, we report the complete polyprotein sequence of a dengue virus 2 strain isolated in brazil. this virus belongs to the american genotype and has the ability to cause neurovirulence in immunocompetent adult mice. the data presented here may help understand the genetic determinants responsible for neurovirulence.201526184939
antiviral activity of chloroquine against dengue virus type 2 replication in aotus monkeys.dengue virus (denv) of the flaviviridae family is a single positive-stranded rna virus that is transmitted by aedes aegypti and aedes albopictus mosquitoes. the objective of this study was to investigate the use of chloroquine (clq) as an antiviral drug against dengue virus in monkeys. to analyze the action of the drug in vivo, nonhuman primates groups (aotus azarai infulatus) were inoculated with a subcutaneous injection of a virulent strain of denv-2, treated and untreated clq. blood hematolog ...201525664975
a small molecule inhibitor of dengue virus type 2 protease inhibits the replication of all four dengue virus serotypes in cell culture.dengue has emerged as the most significant of arboviral diseases in the 21st century. it is endemic to >100 tropical and sub-tropical countries around the world placing an estimated 3.6 billion people at risk. it is caused by four genetically similar but antigenically distinct, serotypes of dengue viruses. there is neither a vaccine to prevent nor a drug to treat dengue infections, at the present time. the major objective of this work was to explore the possibility of identifying a small molecul ...201525886260
emergence of a new lineage of dengue virus type 2 identified in travelers entering western australia from indonesia, 2010-2012.dengue virus (denv) transmission is ubiquitous throughout the tropics. more than 70% of the current global dengue disease burden is borne by people who live in the asia-pacific region. we sequenced the e gene of denv isolated from travellers entering western australia between 2010-2012, most of whom visited indonesia, and identified a diverse array of denv1-4, including multiple co-circulating viral lineages. most viruses were closely related to lineages known to have circulated in indonesia for ...201525635775
clinical outcome and genetic differences within a monophyletic dengue virus type 2 population.the exact mechanisms of interplay between host and viral factors leading to severe dengue are yet to be fully understood. even though previous studies have implicated specific genetic differences of dengue virus (denv) in clinical severity and virus attenuation, similar studies with large-scale, whole genome screening of monophyletic virus populations are limited. therefore, in the present study, we compared 89 whole genomes of denv-2 cosmopolitan clade iii isolates obtained from patients diagno ...201525811657
honokiol, a lignan biphenol derived from the magnolia tree, inhibits dengue virus type 2 infection.dengue is the most widespread arbovirus infection and poses a serious health and economic issue in tropical and subtropical countries. currently no licensed vaccine or compounds can be used to prevent or manage the severity of dengue virus (denv) infection. honokiol, a lignan biphenol derived from the magnolia tree, is commonly used in eastern medicine. here we report that honokiol has profound antiviral activity against serotype 2 denv (denv-2). in addition to inhibiting the intracellular denv- ...201526378567
replacement of pr gene with japanese encephalitis virus pr using reverse genetics reduces antibody-dependent enhancement of dengue virus 2 infection.severe dengue is more likely found during secondary heterologous dengue virus (denv) infection or primary infection of infants born to dengue-immune mothers and led to the hypothesis of antibody-dependent enhancement (ade). it has been reported that pre-membrane (prm)-reactive antibodies do not efficiently neutralize denv infection but instead potently promote ade infection. meanwhile, these enhancing anti-prm antibodies mainly react with the precursor (pr) peptide. to evaluate the effect of pr ...201526219500
the protein diiic-2, aggregated with a specific oligodeoxynucleotide and adjuvanted in alum, protects mice and monkeys against denv-2.previously, we reported the ability of the chimeric protein diiic-2 (domain iii of the dengue envelope protein fused to the capsid protein of dengue-2 virus), to induce immunity and protection in mice, when it is highly aggregated with a non-defined oligodeoxynucleotide (odn) and adjuvanted in alum. in this work, three different defined odns were studied as aggregating agents. our results suggest that the nature of the odn influences the capacity of protein diiic-2 to activate cell-mediated immu ...201525178969
correction: ifi6 inhibits apoptosis via mitochondrial-dependent pathway in dengue virus 2 infected vascular endothelial cells. 201526393359
losartan and enalapril decrease viral absorption and interleukin 1 beta production by macrophages in an experimental dengue virus infection.the role of angiotensin ii (ang ii) in dengue virus infection remains unknown. the aim of this study was to determine the effect of losartan, an antagonist of the angiotensin ii type 1 receptor (at1 receptor), and enalapril, an inhibitor of angiotensin i-converting enzyme (ace), on viral antigen expression and il-1β production in peritoneal macrophages infected with dengue virus type 2. mice treated with losartan or enalapril and untreated controls were infected intraperitoneally with the virus, ...201526321474
the heterogeneous nuclear ribonucleoprotein k (hnrnp k) is a host factor required for dengue virus and junín virus multiplication.heterogeneous nuclear ribonucleoproteins (hnrnps) are cellular factors involved in the replication of several viruses. in this study we analyzed the expression and intracellular localization of hnrnp a2 and hnrnp k in cell cultures infected with two viruses that cause human hemorrhagic fevers: dengue virus type 2 (denv-2) and junín virus (junv). we determined that denv-2 promoted the cytoplasmic translocation of hnrnp k and to a lesser extent of hnrnp a2, meanwhile, junv infection induced an inc ...201525865411
a novel tetravalent formulation combining the four aggregated domain iii-capsid proteins from dengue viruses induces a functional immune response in mice and monkeys.our group developed a subunit vaccine candidate against dengue virus based on two different viral regions: the domain iii of the envelope protein and the capsid protein. the novel chimeric protein from dengue-2 virus [domain iii-capsid (diiic-2)], when presented as aggregated incorporating oligodeoxynucleotides, induced anti-viral and neutralizing antibodies, a cellular immune response and conferred significant protection to mice and monkeys. the remaining constructs were already obtained and pr ...201525795768
nictaba and uda, two glcnac-binding lectins with unique antiviral activity profiles.this study aimed to assess the antiviral properties of a unique lectin (nictaba) produced by the tobacco plant, nicotiana tabacum.201525700718
antigen production using heterologous expression of dengue virus-2 non-structural protein 1 (ns1) in nicotiana tabacum (havana) for immunodiagnostic purposes.expression of dengue-2 virus ns1 protein in nicotiana tabacum plants for development of dengue immunodiagnostic kits. dengue is one of the most important diseases caused by arboviruses in the world. a significant increase in its geographical distribution has been noticed over the last 20 years, with continuous transmission of several serotypes and emergence of the hemorrhagic fever in areas where the disease was previously not prevalent. although the methodological processes for dengue diagnosis ...201525689887
diversity and evolution of the envelope gene of dengue virus type 1 circulating in india in recent times.dengue viral attacks have been reported in various parts of india in recent years. in this paper we report on our studies of the characterisation and evolutionary aspects of gene sequences of the envelope glycoprotein of the prevalent indian dengue virus type 1. comparison with sequences from other countries shows that the envelope genes identified in india are closely related to strains from malaysia. from the evolutionary point of view the envelope gene sequences of this dengue virus of india ...201526642358
synthesis and molecular modelling studies of novel sulphonamide derivatives as dengue virus 2 protease inhibitors.development of antivirals for dengue is now based on rational approach targeting the enzymes involved in its life cycle. among the targets available for inhibition of dengue virus, non-structural protein ns2b-ns3 protease is considered as a promising target for the development of anti-dengue agents. in the current study we have synthesized a series of 4-(1,3-dioxo-2,3-dihydro-1h-isoindol-2-yl)benzene-1-sulphonamide derivatives and screened for denv2 protease activity. compounds 16 and 19 showed ...201526247308
retrospective screening of acute undifferentiated fever serum samples with universal flavivirus primers.fever is a common symptom of many tropical diseases and in many cases the etiologic agent remains unidentified as a consequence of either the etiologic agent not being part of routine diagnostic screening or as a consequence of false negatives on standard diagnostic tests.201526230127
binding specificity of polypeptide substrates in ns2b/ns3pro serine protease of dengue virus type 2: a molecular dynamics study.the pathogenic dengue virus (dv) is a growing global threat, particularly in south east asia, for which there is no specific treatment available. the virus possesses a two-component (ns2b/ns3) serine protease that cleaves the viral precursor proteins. here, we performed molecular dynamics simulations of the ns2b/ns3 protease complexes with six peptide substrates (capsid, intns3, 2a/2b, 4b/5, 3/4a and 2b/3 containing the proteolytic site between p(1) and p(1)' subsites) of dv type 2 to compare th ...201526086900
protein-protein interactions between a. aegypti midgut and dengue virus 2: two-hybrid screens using the midgut cdna library.dengue virus (denv) is principally transmitted by the aedes aegypti mosquito. to date, mosquito population control remains the key strategy for reducing the continuing spread of denv. the focus on the development of new vector control strategies through an understanding of the mosquito-virus relationship is essential, especially targeting the midgut, which is the first mosquito organ exposed to denv infection.201526719940
[expression of dengue virus type 2 nonstructural protein 3 and isolation of host proteins interacting with it].to construct the plasmid expressing the fusion protein of dengue virus type 2 (denv2) nonstructural protein 3 (ns3) with affinity tag, and isolate the cellular proteins interacting with ns3 protein using tandem affinity purification (tap) assay.201526648287
suppression of drug resistance in dengue virus.dengue virus is a major human pathogen responsible for 400 million infections yearly. as with other rna viruses, daunting challenges to antiviral design exist due to the high error rates of rna-dependent rna synthesis. indeed, treatment of dengue virus infection with a nucleoside analog resulted in the expected genetic selection of resistant viruses in tissue culture and in mice. however, when the function of the oligomeric core protein was inhibited, no detectable selection of drug resistance i ...201526670386
feasibility of using the mosquito blood meal for rapid and efficient human and animal virus surveillance and discovery.mosquito blood meals taken from humans and animals potentially represent a useful source of blood for the detection of blood-borne pathogens. in this feasibility study, anopheles stephensi mosquitoes were fed with blood meals spiked with dengue virus type 2 (denv-2) and harvested at serial time points. these mosquitoes are not competent vectors, and the virus is not expected to replicate. ingested blood was spotted on whatman fta cards and stored at room temperature. mosquito abdomens were remov ...201526416112
the pattern of adipose tissue accumulation during early infancy provides an environment for the development of dengue hemorrhagic fever.dengue is the most prevalent arthropod-borne viral illness in humans with half of the world's population at risk. during early infancy, severe dengue can develop after a primary dengue virus infection. there has been a clinical observation that severe dengue during the first year of life is seen only in chubby infants.201526636570
viral small-rna analysis of bombyx mori larval midgut during persistent and pathogenic cytoplasmic polyhedrosis virus infection.the lepidopteran innate immune response against rna viruses remains poorly understood, while in other insects several studies have highlighted an essential role for the exo-rnai pathway in combating viral infection. here, by using deep-sequencing technology for viral small-rna (vsrna) assessment, we provide evidence that exo-rnai is operative in the silkworm bombyx mori against both persistent and pathogenic infection of b. mori cytoplasmic polyhedrosis virus (bmcpv) which is characterized by a ...201526339065
hybrid flagellin as a t cell independent vaccine scaffold.to extend the potency of vaccines against infectious diseases, vaccines should be able to exploit multiple arms of the immune system. one component of the immune system that is under-used in vaccine design is the subset of b cells known to be capable of responding to repetitive antigenic epitopes and differentiate into plasma cells even in the absence of t cell help (t-independent, ti).201526265529
nmr and md studies reveal that the isolated dengue ns3 protease is an intrinsically disordered chymotrypsin fold which absolutely requests ns2b for correct folding and functional dynamics.dengue genome encodes a two component protease complex (ns2b-ns3pro) essential for the viral maturation/infectivity, thus representing a key drug target. previously, due to its "complete insolubility", the isolated ns3pro could not be experimentally studied and it remains elusive what structure it adopts without ns2b and why ns2b is indispensable. here as facilitated by our previous discovery, the isolated ns3pro has been surprisingly deciphered by nmr to be the first intrinsically-disordered ch ...201526258523
rna viruses and rnai: quasispecies implications for viral escape.due to high mutation rates, populations of rna viruses exist as a collection of closely related mutants known as a quasispecies. a consequence of error-prone replication is the potential for rapid adaptation of rna viruses when a selective pressure is applied, including host immune systems and antiviral drugs. rna interference (rnai) acts to inhibit protein synthesis by targeting specific mrnas for degradation and this process has been developed to target rna viruses, exhibiting their potential ...201526102581
attenuation of dengue (and other rna viruses) with codon pair recoding can be explained by increased cpg/upa dinucleotide frequencies. 201526071447
dengue virus requires the cc-chemokine receptor ccr5 for replication and infection development.dengue is a mosquito-borne disease that affects millions of people worldwide yearly. currently, there is no vaccine or specific treatment available. further investigation on dengue pathogenesis is required to better understand the disease and to identify potential therapeutic targets. the chemokine system has been implicated in dengue pathogenesis, although the specific role of chemokines and their receptors remains elusive. here we describe the role of the cc-chemokine receptor ccr5 in dengue v ...201525939314
a novel iminosugar uv-12 with activity against the diverse viruses influenza and dengue (novel iminosugar antiviral for influenza and dengue).iminosugars are capable of targeting the life cycles of multiple viruses by blocking host endoplasmic reticulum α-glucosidase enzymes that are required for competent replication of a variety of enveloped, glycosylated viruses. iminosugars as a class are approved for use in humans with diseases such as diabetes and gaucher's disease, providing evidence for safety of this class of compounds. the in vitro antiviral activity of iminosugars has been described in several publications with a subset of ...201525984714
hcv induces the expression of rubicon and uvrag to temporally regulate the maturation of autophagosomes and viral replication.hepatitis c virus (hcv) induces autophagy to enhance its replication. however, how hcv regulates the autophagic pathway remains largely unclear. in this report, we demonstrated that hcv infection could induce the expression of rubicon and uvrag, which inhibited and stimulated the maturation of autophagosomes, respectively. the induction of rubicon by hcv was prompt whereas the induction of uvrag was delayed, resulting in the accumulation of autophagosomes in the early time points of viral infect ...201525807108
virocidal activity of egyptian scorpion venoms against hepatitis c virus.hepatitis c virus (hcv) is a major global health problem, causing chronic hepatitis, liver cirrhosis and hepatocellular carcinoma. development of well-tolerated regimens with high cure rates and fewer side effects is still much needed. recently, natural antimicrobial peptides (amps) are attracting more attention as biological compounds and can be a good template to develop therapeutic agents, including antiviral agents against a variety of viruses. various amps have been characterized from the v ...201525889296
computational prediction and analysis of envelop glycoprotein epitopes of denv-2 and denv-3 pakistani isolates: a first step towards dengue vaccine development.dengue fever of tropics is a mosquito transmitted devastating disease caused by dengue virus (denv). there is no effective vaccine available, so far, against any of its four serotypes (denv-1, denv-2, denv-3, and denv-4). there is a need for the development of preventive and therapeutic vaccines against denv to decrease the prevalence of dengue fever, especially in pakistan. in this research, linear and conformational b-cell epitopes of envelope glycoprotein of denv-2 and denv-3 (the most preval ...201525775090
dengue patients exhibit higher levels of prm and e antibodies than their asymptomatic counterparts.dengue virus infection is a common tropical disease which often occurs without being detected. these asymptomatic cases provide information in relation to the manifestation of immunological aspects. in this study, we developed an elisa method to compare neutralizing effects of dengue prm and e antibodies between dengue patients and their asymptomatic household members. recombinant d2 premembrane (prm) was constructed, cloned, and tested for antigenicity. the recombinant protein was purified and ...201525815314
the effect of hydrocotyle sibthorpioides lam. extracts on in vitro dengue replication.objective. to investigate the potential effect of hydrocotyle sibthorpioides lam. (h. sibthorpioides) extracts against in vitro dengue viral replication. methods. the cytotoxicity of h. sibthorpioides was evaluated using a cell viability assay. cells were pre- and posttreated with water and methanol extracts of h. sibthorpioides, and the viral inhibitory effect was investigated by observing the morphological changes, which were further confirmed by plaque assay. results. the methanolic extract c ...201525767554
hela nucleic acid contamination in the cancer genome atlas leads to the misidentification of human papillomavirus 18.we searched the cancer genome atlas (tcga) database for viruses by comparing non-human reads present in transcriptome sequencing (rna-seq) and whole-exome sequencing (wxs) data to viral sequence databases. human papillomavirus 18 (hpv18) is an etiologic agent of cervical cancer, and as expected, we found robust expression of hpv18 genes in cervical cancer samples. in agreement with previous studies, we also found hpv18 transcripts in non-cervical cancer samples, including those from the colon, r ...201525631090
flavivirus sfrna suppresses antiviral rna interference in cultured cells and mosquitoes and directly interacts with the rnai machinery.productive arbovirus infections require mechanisms to suppress or circumvent the cellular rna interference (rnai) pathway, a major antiviral response in mosquitoes. in this study, we demonstrate that two flaviviruses, dengue virus and kunjin virus, significantly repress sirna-mediated rnai in infected human cells as well as during infection of the mosquito vector culex quinquefasciatus. arthropod-borne flaviviruses generate a small structured non-coding rna from the viral 3' utr referred to as s ...201526331679
the interferon-inducible mouse apolipoprotein l9 and prohibitins cooperate to restrict theiler's virus replication.apolipoprotein l9b (apol9b) is an interferon-stimulated gene (isg) that has antiviral activity and is weakly expressed in primary mouse neurons as compared to other cell types. here, we show that both apol9 isoforms (apol9b and apol9a) inhibit replication of theiler's murine encephalomyelitis virus (tmev) but not replication of vesicular stomatitis virus (vsv), murid herpesvirus-4 (muhv-4), or infection by a lentiviral vector. apol9 genes are strongly expressed in mouse liver and, to a lesser ex ...201526196674
xrn1 stalling in the 5' utr of hepatitis c virus and bovine viral diarrhea virus is associated with dysregulated host mrna stability.we demonstrate that both hepatitis c virus (hcv) and bovine viral diarrhea virus (bvdv) contain regions in their 5' utrs that stall and repress the enzymatic activity of the cellular 5'-3' exoribonuclease xrn1, resulting in dramatic changes in the stability of cellular mrnas. we used biochemical assays, virus infections, and transfection of the hcv and bvdv 5' untranslated regions in the absence of other viral gene products to directly demonstrate the existence and mechanism of this novel host-v ...201525747802
standing your ground to exoribonucleases: function of flavivirus long non-coding rnas.members of the flaviviridae (e.g., dengue virus, west nile virus, and hepatitis c virus) contain a positive-sense rna genome that encodes a large polyprotein. it is now also clear most if not all of these viruses also produce an abundant subgenomic long non-coding rna. these non-coding rnas, which are called subgenomic flavivirus rnas (sfrnas) or xrn1-resistant rnas (xrrnas), are stable decay intermediates generated from the viral genomic rna through the stalling of the cellular exoribonuclease ...201526368052
spot the difference-development of a syndrome based protein microarray for specific serological detection of multiple flavivirus infections in travelers.the family flaviviridae, genus flavivirus, holds many of the world's most prevalent arboviral diseases that are also considered the most important travel related arboviral infections. in most cases, flavivirus diagnosis in travelers is primarily based on serology as viremia is often low and typically has already been reduced to undetectable levels when symptoms set in and patients seek medical attention. serological differentiation between flaviviruses and the false-positive results caused by va ...201525767876
usutu virus: an emerging flavivirus in europe.usutu virus (usuv) is an african mosquito-borne flavivirus belonging to the japanese encephalitis virus serocomplex. usuv is closely related to murray valley encephalitis virus, japanese encephalitis virus, and west nile virus. usuv was discovered in south africa in 1959. in europe, the first true demonstration of circulation of usuv was reported in austria in 2001 with a significant die-off of eurasian blackbirds. in the subsequent years, usuv expanded to neighboring countries, including italy, ...201525606971
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