Molecular epidemiology of dengue viruses from complete genome sequences inauguraldissertation zur. At the same time, significant advances in genomics approaches have provided insights into hostpathogen. Genomics, proteomics and evolution of dengue virus oxford. Intact platelets squares or platelets subjected to 3 cycles of freezethaw at. Molecular characterization of dengue viruses circulating. Whole genome sequencing and genetic variations in several dengue virus type 1 strains from unusual dengue epidemic of 2017 in vietnam. In some cases, antidenguevirus igm can be undetectable.
Notably, the multifunctional ns1 protein readily tolerated insertions in regions within the wing, connector, and. The disease is caused by four serologically and genetically distinct viruses termed dengue virus denv 1, 2, 3 and 4. Complete genome sequences of dengue virus type 1 to 4. Dengue virus denv is a member of the flavivirus genus of the flaviviridae family. In june 2017, dengue virus type 2 infection was diagnosed in 2 travelers returned to japan from sri lanka, where the countrys largest dengue fever outbreak is ongoing. Dengue virus denv infection is the most common mosquitotransmitted viral infection. Pervasive tertiary structure in the dengue virus rna genome. It is a mosquitoborne, single positivestranded rna virus of the family flaviviridae.
Sep 18, 2015 a new understanding of dengue virus date. Dengue virus denv and chikungunya virus chikv are arboviruses that share the same aedes mosquito vectors and thus overlap in their endemic areas. Dengue is a major public health problem worldwide, where an estimated 400 million denv infections and 100 million clinically apparent dengue cases occurred in 2010. Sep 03, 2015 by successfully editing specific genes in the aedes aegytpi, the mosquito species which transmits the dengue virus, we have established techniques which can be used in future research to target. Travelers, especially those previously affected by dengue fever, should take measures to avoid mosquito bites. Dengue virus rna structure specialization facilitates host.
Isolation and complete genome analysis of neurotropic. Rna viruses usurp and reprogram host cells using short rna genomes. Dengue viruses denvs are mosquitoborne viruses which can cause. Largescale recoding of an arbovirus genome to rebalance. Denv has a singlestranded, positivesense rna genome that is approximately 11 kb in length, which. Flaviviruses are enveloped viruses with a single stranded. In 2015, a dengue outbreak with 1067 reported cases occurred in.
Molecular characterization of dengue virus serotype 2. The cocirculation of zika virus and dengue virus in. Chikungunya virus and the global spread of a mosquitoborne disease. Virus pathogen database and analysis resource vipr. Dengue virus denv of flaviviridae family has emerged as the. Deep sequencing of the dengue virus 1 genome directly from the blood of twelve dengue patients and from mosquitoes given this blood showed consistent and distinct mutation patterns during infection. Scientists successfully edit genes of dengue fever mosquitoes. Laboratory dengue diagnosis can be performed through virus isolation, genome and antigen detection and serological studies. Denv has a singlestranded, positivesense rna genome that is approximately 11 kb in length. Onestep realtime rtpcr assays for serotyping dengue. I n 2009, sri lanka experienced an outbreak of dengue. Research paper insilico designing and testing of primers. Analysis of dengue virus genetic diversity during human and mosquito infection reveals genetic constraints.
Denv genome was not detected in plasma or peripheral blood mononuclear cells pbmc samples on day 7, 14 or 21. Given the geographic expansion of denv14, assays for serotyping are needed to be able to perform surveillance and epidemiological studies. We use a new highthroughput rna structure probing technology to. Dengue viruses are spread to people through the bite of an infected aedes species ae. Zika virus in the americasyet another arbovirus threat. In secondary infections, antidenguevirus igg evolves rapidly, with high titres soon after fever onset. Dengue viruses have spread rapidly within countries and across regions in the past few decades, resulting in an increased frequency of epidemics and severe dengue disease, hyperendemicity of multiple dengue virus serotypes in many tropical countries, and autochthonous transmission in europe and the usa. Complete genome sequence and phylogenetic analysis.
Dengue, one of the most devastating mosquitoborne viral diseases in humans, is a significant life threatening problem in india and other developing countries. Interplay of dengue virus and the human immune response the harvard community has made this article openly available. The present study was planned to identify the serotypegenotype of dengue viruses circulating in uttar pradesh, india. Dengue is a potentially fatal acute febrile illness caused by infection with any of four dengue viruses denv1, 2, 3, and 4. Dengue is often a leading cause of illness in areas. Complete genome sequence of an atypical dengue virus. Dengue is among the most important mosquitoborne viral disease in the world. Denv infection can cause mild dengue fever or severe dengue hemorrhagic fever dhfdengue shock syndrome dss. Nov 06, 2018 rna viruses usurp and reprogram host cells using short rna genomes. An international consortium of scientists worked to map out the. After washing 3 times and resuspending the platelets in serumfree m199, the pellets were cultured at.
Dengue viruses are members of the family flaviviridae genus flavivirus. Functional analysis of the dengue virus genome using an. Functional rna elements in the dengue virus genome. However, the complete genome sequences of denv2 isolates imported from myanmar are not available. Fulllength genome and molecular characterization of. In this study, we describe the design and validation. Antidengue virus denv igm and igg were identified in plasma samples on days 7, 14 and 25 after the onset of symptoms. For virus detection, dengue viremia is short, usually observed two or three days before onset of fever and lasts four to five days later. The viral genomes resource is a collection of viral genomic sequences that is a part of the entrez genomes, which provides curated sequence data and annotations of complete genomes to the scientific community. Dengue viral infections are known to pose a significant risk during travel to tropical regions, but it is surprising to find dengue transmission in the hilly region of nepal, which is over 1800mtr above sea level. Analysis of dengue virus genetic diversity during human and. Dengue virus is the leading cause of arboviral disease in the world, with more than 390 million casesyear in tropical and subtropical regions of the americas, asia, africa, and pacific islands. A retrospective study found no evidence that yellow fever vaccination in dengueendemic areas increases risk for severe dengue fever.
The mechanisms involved in dengue pathogenesis and severity of the symptoms vary according to host and viral factors. Denvns1 positive serum samples were collected from dengue fever patients, and complete genome sequences were obtained. Author summary dengue viruses denv are viruses that can cause asymptomatic infection to lifethreatening haemorrhagic fever disease. In order to characterize the denv1jeddah12011 isolate, the full genome was sequenced.
Jan 16, 2015 complete genome sequence and phylogenetic analysis. Genomewide mutagenesis of dengue virus reveals plasticity of. Largescale recoding of an arbovirus genome to rebalance its insect versus mammalian preference sam h. These mosquitoes are also vectors of chikungunya, yellow fever and zika viruses. Dengue is the most common mosquitoborne infection worldwide and a serious threat to global public health. Primers for dengue virus strains based on their sequence. The assay was performed according to the operation manual. The authors show that platelets synthesize ns1 protein and.
Denvns1 positive serum samples were collected from dengue fever patients, and complete genome sequences. To characterize the virus, the complete genome sequence was obtained and phylogenetic. Laboratory diagnosis is also important for case confirmation. Complete genome sequencing and phylogenetic analysis of. These samples were examined for virus genome by realtime rtpcr taqman rtpcr ito et al.
Screening the dengue virus genome for tolerated insertions. Dengue virus denv infection is responsible for the most significant mosquitoborne viral disease in the world today. Since the dengue virus was first isolated in 1943, studies to understand the pathogenesis of dengue has long been hindered by the lack of suitable experimental models. Each denv has four or more genetic groups or genotypes. By successfully editing specific genes in the aedes aegytpi, the mosquito species which transmits the dengue virus, we have established techniques which can be. Plateletdependent replication of the dengue virus genome. Reviews highlight new ideas and directions in basic virology, viral disease mechanisms, virushost interactions, and cellular and immune responses to virus infection, and reinforce the position of viruses as uniquely powerful probes of. Unlike the other flaviviruses, there are 4 serotypes, referred to as denv14, that are genetically similar but antigenically. Today, dengue is regarded as the most prevalent and rapidly spreading mosquitoborne viral. Villordo sm, filomatori cv, sanchezvargas i, blair cd, gamarnik av 2015 dengue virus rna structure specialization facilitates.
Hemorrhage and vascular leakage are two characteristic symptoms of dhfdss. It is an endemic arbovirus affecting more than 390 million people worldwide. In the last 50 years, dengue incidence has increased 30fold, worldwide. Dengue virus denv belongs to the genus flavivirus in the family flaviviridae. Genome wide analyses reveal a highly conserved dengue virus envelope peptide which is critical for virus viability and antigenic in humans. In 2015, a dengue outbreak with 1,067 reported cases occurred in xishuangbanna, a city in china that borders burma and laos. Zika virus might show crossreactivity with other flaviviruses and possibly cytomegalovirus, and hematologic disorders might cause ns1 positivity 6,7. They are small enveloped viruses containing a singlestrand rna genome of positive polarity. Dengue virus denv is an enveloped virus with a singlestranded. There are four dengue serotypes denv1 to denv4, which are transmitted by aedes mosquitoes, mainly a. Staufta,b,1, oleksandr gorbatsevycha, yutong songa, charles b. Importantly, we identified regions within the viral genome that are resistant to mutation during human and mosquito infection.
Dengue virus infection in a french traveller to the hilly. To characterize the virus, the complete genome sequence was obtained and phylogenetic, mutation, substitution and recombinant analyses were performed. Bernke te winkel, christof schaefer, in drugs during pregnancy and lactation third edition, 2015. Interplay of dengue virus and the human immune response. Here we report the complete genome sequences of these three laboratory strains, dengue virus. Aug 20, 2018 dengue is the most common mosquitoborne infection worldwide and a serious threat to global public health.
Dengue virus binding and replication by platelets blood. Rna viruses encode the information required for their replication in both their primary sequences and higherorder structures formed when the rna genome strand folds back on itself, but the extent of higherorder structure has remained unclear. The authors show that platelets synthesize ns1 protein and replicate the dengue viral genome. Dengue is common in more than 100 countries around the world. Genomewide analyses reveal a highly conserved dengue. Deep sequencing of the dengue virus 1 genome directly from the blood of. The fda announced approval of dengvaxia vaccine for prevention of dengue disease caused by all dengue serotypes in people ages 916 who have laboratoryconfirmed prior infection and live in endemic areas.
To characterize the virus, the complete genome sequence was obtained. Of 433 dengue suspected patients, tested by reverse transcriptase pcr rt. Tracking dengue virus intrahost genetic diversity during humantomosquito transmission. Dengue is a mosquitoborne viral disease that has rapidly spread in all regions of who in recent years. Jun 27, 2018 in 2015, a dengue outbreak with 1,067 reported cases occurred in xishuangbanna, a city in china that borders burma and laos. Molecular epidemiology of dengue viruses from complete.
Sporadic dengue virus serotype 2 denv2 imported cases from myanmar have been documented almost every year in yunnan province of china since 2005. Dengue virus binding and replication by platelets blood american. Wardc,2, alisa yurovskya, steffen muellera,b, bruce futchera, and eckard wimmera,b,3 adepartment of molecular genetics and microbiology and cdepartment of computer science, stony brook university school of. The flavivirus genus includes other important human pathogens such as yellow fever yfv, west nile wnv, japanese encephalitis jev, and tick borne encephalitis tbev. Dengue virus pirates human platelets blood american. During designing and selection of primers these complexities needs to be over ruled. Dengue virus perturbs mitochondrial morphodynamics to. The genome of flaviviruses is approximately 11 000 bases long and they share. Five serotypes of the virus have been found, all of which can cause the full spectrum of disease. Climate change, rapid urbanization and increased global travel have facilitated the spread of both efficient mosquito vectors and the four dengue virus serotypes between population centers. Dengue virus is transmitted by female mosquitoes mainly of the species aedes aegypti and, to a lesser extent, ae. However, due to the limited understanding of dengue pathogenesis, no satisfactory therapies to treat.
Complete genome sequences of three laboratory strains of. Three of these are structural proteins that form the coat of the virus and deliver the rna to target cells, and seven of them are nonstructural proteins that orchestrate the production of new viruses once the virus gets inside the cell. A computational method for the identification of dengue. More than 50% of the viruses in the genus flavivirus, family flaviviridae, cause disease in humans, including dengue, yellow fever, west nile and zika viruses. Nevertheless, scientists understanding of dengue virus may be simplistic, as rather than distinct antigenic groups, a continuum appears to exist. Considering the challenges of an appropriate primer design for pcr based sequencing of whole genome of dengue virus subtypes, manyof the pcrbased whole genome sequencing methods, as reported in the literature. Because the outcomes and management strategies for. A 43yearold caucasian female traveler from france presented with fever and abdominal pain following a diarrheal illness while visiting the central hilly region of nepal.
Dengue virus is a small virus that carries a single strand of rna as its genome. These two viruses also cause similar clinical presentations, especially in the initial stages of infection, with neither virus possessing any specific distinguishing clinical features. Genomewide analyses reveal a highly conserved dengue virus envelope peptide which is critical for virus viability and antigenic in humans. A risk to anyone traveling to a tropical or subtropical region of the world where dengue virus is found. Complete genome sequencing and phylogenetic analysis of dengue. Assembly of all sequenced overlapping fragments obtained by rtpcr resulted in a genome of 10,622 nts with a complete coding region encoding for 3392 amino acids aa flanked by 94 and 349 nts at the 5 utr and 3 utr, respectively. Dengue and severe dengue world health organization. Analysis of dengue virus genetic diversity during human. The incidence and geographic range of dengue have increased dramatically in recent decades. Like other flaviviruses, its genome comprises a single strand of positivesense rna encoding 3 structural and 7 nonstructural ns proteins.
Although denv are not classically known to infect and invade central nervous system cns in human, numerous cases of denv infection in the cns have been reported with limited information about the characteristics of the infecting virus. Dengue virus type 2 in travelers returning to japan from. Dengue viruses denv cause debilitating and potentially. University of texas medical branch at galveston summary. Whole genome sequence alignment and phylogenetic analysis showed high. Report rigi recognizes the 50 region of dengue and zika virus genomes graphical abstract highlights d an af.
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