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the first isolation of bussuquara virus from man. 19715107116
identification of brazilian flaviviruses by a simplified reverse transcription-polymerase chain reaction method using flavivirus universal primers.we report a simplified reverse transcription-polymerase chain reaction (rt-pcr) method for identification of brazilian flaviviruses based on the patterns of electrophoretic separation of the amplicons. the rt-pcr was done on the culture fluids of aedes albopictus c6/36 cells infected with brazilian flaviviruses, without previous extraction of viral rna, using flavivirus universal primers that anneal to highly conserved sequences within the nonstructural protein 5 and 3'- non translated region of ...19989749625
epitope-blocking enzyme-linked immunosorbent assays for detection of west nile virus antibodies in domestic mammals.we evaluated the ability of epitope-blocking enzyme-linked immunosorbent assays (elisas) to detect west nile virus (wnv) antibodies in domestic mammals. sera were collected from experimentally infected horses, cats, and pigs at regular intervals and screened in elisas and plaque reduction neutralization tests. the diagnostic efficacies of these techniques were similar.200312791902
serologic evidence of west nile virus infection in horses, yucatan state, mexico.serum samples were obtained from 252 horses in the state of yucatan, mexico, from july to october 2002. antibodies to west nile virus were detected by epitope-blocking enzyme-linked immunosorbent assays in three (1.2%) horses and confirmed by plaque reduction neutralization test. we report the first west nile virus activity in the state of yucatan.200312890328
west nile virus antibodies in colombian horses. 200516673523
development of multiplex real-time reverse transcriptase pcr assays for detecting eight medically important flaviviruses in mosquitoes.a multiplex real-time reverse transcriptase pcr has been developed for the rapid detection and identification of eight medically important flaviviruses from laboratory-reared, virus-infected mosquito pools. the method used involves the gene-specific amplification of yellow fever virus (yfv), japanese encephalitis virus (jev), west nile virus (wnv), st. louis encephalitis virus (slev), and dengue virus (denv) serotypes 1 to 4 (denv-1 to denv-4, respectively) by use of the flavivirus consensus amp ...200717108075
constancy and diversity in the flavivirus fusion peptide.flaviviruses include the mosquito-borne dengue, japanese encephalitis, yellow fever and west nile and the tick-borne encephalitis viruses. they are responsible for considerable world-wide morbidity and mortality. viral entry is mediated by a conserved fusion peptide containing 16 amino acids located in domain ii of the envelope protein e. highly orchestrated conformational changes initiated by exposure to acidic ph accompany the fusion process and are important factors limiting amino acid change ...200818275613
detection of rna from a novel west nile-like virus and high prevalence of an insect-specific flavivirus in mosquitoes in the yucatan peninsula of mexico.as part of our ongoing surveillance efforts for west nile virus (wnv) in the yucatan peninsula of mexico, 96,687 mosquitoes collected from january through december 2007 were assayed by virus isolation in mammalian cells. three mosquito pools caused cytopathic effect. two isolates were orthobunyaviruses (cache valley virus and kairi virus) and the identity of the third infectious agent was not determined. a subset of mosquitoes was also tested by reverse transcription-polymerase chain reaction (r ...200919141845
characterization of a novel flavivirus from mosquitoes in northern europe that is related to mosquito-borne flaviviruses of the tropics.a novel flavivirus was isolated from mosquitoes in finland, representing the first mosquito-borne flavivirus from northern europe. the isolate, designated lammi virus (lamv), was antigenically cross-reactive with other flaviviruses and exhibited typical flavivirus morphology as determined by electron microscopy. the genomic sequence of lamv was highly divergent from the recognized flaviviruses, and yet the polyprotein properties resembled those of mosquito-borne flaviviruses. phylogenetic analys ...200919570865
virus-host coevolution: common patterns of nucleotide motif usage in flaviviridae and their hosts.virus-host biological interaction is a continuous coevolutionary process involving both host immune system and viral escape mechanisms. flaviviridae family is composed of fast evolving rna viruses that infects vertebrate (mammals and birds) and/or invertebrate (ticks and mosquitoes) organisms. these host groups are very distinct life forms separated by a long evolutionary time, so lineage-specific anti-viral mechanisms are likely to have evolved. flaviviridae viruses which infect a single host l ...200919617912
structure of yellow fever virus envelope protein domain iii.the structure of recombinant domain iii of the envelope protein (red3) of yellow fever virus (yfv), containing the major neutralization site, was determined using nmr spectroscopy. the amino acid sequence and structure of the yfv-red3 shows differences from ed3s of other mosquito-borne flaviviruses; in particular, the partially surface-exposed bc loop where methionine-304 and valine-324 were identified as being critical for the structure of the loop. variations in the structure and surface chemi ...200919818466
evaluation of two molecular methods for the detection of yellow fever virus genome.yellow fever virus (yfv), a member of the family flaviviridae, genus flavivirus is endemic to tropical areas of africa and south america and is among the arboviruses that pose a threat to public health. recent outbreaks in brazil, bolivia, and paraguay and the observation that vectors capable of transmitting yfv are presenting in urban areas underscore the urgency of improving surveillance and diagnostic methods. two novel methods (rt-hemi-nested-pcr and sybr(®) green qrt-pcr) for efficient dete ...201121419803
duck egg-drop syndrome caused by byd virus, a new tembusu-related flavivirus.since april 2010, a severe outbreak of duck viral infection, with egg drop, feed uptake decline and ovary-oviduct disease, has spread around the major duck-producing regions in china. a new virus, named byd virus, was isolated in different areas, and a similar disease was reproduced in healthy egg-producing ducks, infecting with the isolated virus. the virus was re-isolated from the affected ducks and replicated well in primary duck embryo fibroblasts and vero cells, causing the cytopathic effec ...201121455312
a diagnostic algorithm to serologically differentiate west nile virus from japanese encephalitis virus infections and its validation in field surveillance of poultry and horses.abstract the detection of west nile virus (wnv) in areas endemic for japanese encephalitis virus (jev) is complicated by the extensive serological cross-reactivity between the two viruses. a testing algorithm was developed and employed for the detection of anti-wnv antibody in areas endemic for jev. using this differentiation algorithm, a serological survey of poultry (2004 through 2009) and horses (2007 through 2009) was performed. among 2681 poultry sera, 125 samples were interpreted as bein ...201222217162
serological evidence of flaviviruses and alphaviruses in livestock and wildlife in trinidad.seroprevalence rates of selected arboviruses in animal populations in trinidad were determined using serum samples collected between 2006 and 2009 from horses (n=506), cattle (n=163), sheep (n=198), goats (n=82), pigs (n=184), birds (n=140), rodents (n=116), and other vertebrates (n=23). the sera were screened for antibodies to west nile virus (wnv), st. louis encephalitis virus (slev), ilheus virus (ilhv), bussuquara virus (bsqv), venezuelan equine encephalitis virus (veev), eastern equine ence ...201222989182
biological transmission of arboviruses: reexamination of and new insights into components, mechanisms, and unique traits as well as their evolutionary trends.among animal viruses, arboviruses are unique in that they depend on arthropod vectors for transmission. field research and laboratory investigations related to the three components of this unique mode of transmission, virus, vector, and vertebrate host, have produced an enormous amount of valuable information that may be found in numerous publications. however, despite many reviews on specific viruses, diseases, or interests, a systematic approach to organizing the available information on all f ...016223950
ecuador paraiso escondido virus, a new flavivirus isolated from new world sand flies in ecuador, is the first representative of a novel clade in the genus flavivirus.a new flavivirus, ecuador paraiso escondido virus (epev), named after the village where it was discovered, was isolated from sand flies (psathyromyia abonnenci, formerly lutzomyia abonnenci) that are unique to the new world. this represents the first sand fly-borne flavivirus identified in the new world. epev exhibited a typical flavivirus genome organization. nevertheless, the maximum pairwise amino acid sequence identity with currently recognized flaviviruses was 52.8%. phylogenetic analysis o ...201526355096
new insights into flavivirus evolution, taxonomy and biogeographic history, extended by analysis of canonical and alternative coding sequences.to generate the most diverse phylogenetic dataset for the flaviviruses to date, we determined the genomic sequences and phylogenetic relationships of 14 flaviviruses, of which 10 are primarily associated with culex spp. mosquitoes. we analyze these data, in conjunction with a comprehensive collection of flavivirus genomes, to characterize flavivirus evolutionary and biogeographic history in unprecedented detail and breadth. based on the presumed introduction of yellow fever virus into the americ ...201525719412
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
dna microarray platform for detection and surveillance of viruses transmitted by small mammals and arthropods.viruses transmitted by small mammals and arthropods serve as global threats to humans. most emergent and re-emergent viral agents are transmitted by these groups; therefore, the development of high-throughput screening methods for the detection and surveillance of such viruses is of great interest. in this study, we describe a dna microarray platform that can be used for screening all viruses transmitted by small mammals and arthropods (smaviruschip) with nucleotide sequences that have been depo ...201627654806
mayaro virus and dengue virus 1 and 4 natural infection in culicids from cuiabá, state of mato grosso, brazil.this study aimed to verify the diversity of culicidae species and their frequency of infection with flaviviruses and alphaviruses in cuiabá, state of mato grosso, brazil. mosquitoes were captured with nasci aspirators and hand net in 200 census tracts, identified alive at species level and pooled in one-20 (11,090 mosquitoes, 14 species). female pools (n = 610) were subjected to multiplex seminested-reverse transcription-polymerase chain reaction (rt-pcr) for 11 flavivirus and five alphavirus. p ...026784852
clinical and virological descriptive study in the 2011 outbreak of dengue in the amazonas, brazil.dengue is a vector-borne disease in the tropical and subtropical region of the world and is transmitted by the mosquito aedes aegypti. in the state of amazonas, brazil during the 2011 outbreak of dengue all the four dengue virus (denv) serotypes circulating simultaneously were observed. the aim of the study was to describe the clinical epidemiology of dengue in manaus, the capital city of the state of the amazonas, where all the four denv serotypes were co-circulating simultaneously.201424978469
seroprevalence of flaviviruses antibodies in water buffaloes (bubalus bubalis) in brazilian amazon.the state of pará encompasses 26% of brazilian amazon where an enormous diversity of arboviruses has been found. this study aimed to assess the prevalence and distribution of hemagglutination-inhibition antibodies against antigens of six flavivirus (yellow fever virus, ilheus virus, saint louis encephalitis virus, cacipacore virus, bussuquara virus and rocio virus) in water buffaloes in pará state, brazil. the prevalence of antibodies in these farm animals is important to determine the circulati ...201424666635
serological evidence of widespread exposure of grenada fruit bats to chikungunya virus.antibody detection against selected potentially zoonotic vector-borne alphaviruses and flaviviruses was conducted on sera from bats from all six parishes in grenada, west indies. sera were tested for (i) antibodies to flaviviruses west nile virus, st. louis encephalitis virus, ilhéus virus, bussuquara virus (bsqv), rio bravo virus and all four serotypes of dengue virus (denv) by plaque reduction neutralization test (prnt); (ii) antibodies to alphaviruses western equine encephalitis virus, venezu ...201829575672
flavidb: a data mining system for knowledge discovery in flaviviruses with direct applications in immunology and vaccinology.the flavivirus genus is unusually large, comprising more than 70 species, of which more than half are known human pathogens. it includes a set of clinically relevant infectious agents such as dengue, west nile, yellow fever, and japanese encephalitis viruses. although these pathogens have been studied extensively, safe and efficient vaccines lack for the majority of the flaviviruses.025544857
approaches for the development of rapid serological assays for surveillance and diagnosis of infections caused by zoonotic flaviviruses of the japanese encephalitis virus serocomplex.flaviviruses are responsible for a number of important mosquito-borne diseases of man and animals globally. the short vireamic period in infected hosts means that serological assays are often the diagnostic method of choice. this paper will focus on the traditional methods to diagnose flaviviral infections as well as describing the modern rapid platforms and approaches for diagnostic antigen preparation.201222570528
diagnosis of arboviruses using indirect sandwich igg elisa in horses from the brazilian amazon.the amazon as a whole is the largest reservoir of arboviruses worldwide, while the brazilian amazon hosts the largest variety of arboviruses isolated to date. in this study, the results of an indirect sandwich igg elisa, standardized for 19 arbovirustypes circulating among horses in brazilian amazon, were compared to results of the hemagglutination inhibition test. a screening test assessed the conditional probability distribution and a pearson linear correlation test determined the correlation ...201425013410
hierarchical cluster analysis of three-dimensional reconstructions of unbiased sampled microglia shows not continuous morphological changes from stage 1 to 2 after multiple dengue infections in callithrix penicillata.it is known that microglial morphology and function are related, but few studies have explored the subtleties of microglial morphological changes in response to specific pathogens. in the present report we quantitated microglia morphological changes in a monkey model of dengue disease with virus cns invasion. to mimic multiple infections that usually occur in endemic areas, where higher dengue infection incidence and abundant mosquito vectors carrying different serotypes coexist, subjects receiv ...201627047345
structure and functionality in flavivirus ns-proteins: perspectives for drug design.flaviviridae are small enveloped viruses hosting a positive-sense single-stranded rna genome. besides yellow fever virus, a landmark case in the history of virology, members of the flavivirus genus, such as west nile virus and dengue virus, are increasingly gaining attention due to their re-emergence and incidence in different areas of the world. additional environmental and demographic considerations suggest that novel or known flaviviruses will continue to emerge in the future. nevertheless, u ...201019945487
cytokine and nitric oxide production by mouse macrophages infected with brazilian flaviviruses.the flaviviridae family, flavivirus genus includes viruses that are transmitted to vertebrates by infected mosquitoes or ticks. the genus flavivirus includes a variety of viruses that cause diseases such as acute febrile illness, encephalitis, and hemorrhagic fever. flaviviruses primarily infect blood monocytes and tissue macrophages, which have been shown to be permissive, supporting viral replication and serving as virus reservoirs. on the other hand, these cells may have an important antivira ...200919551288
universal primers that amplify rna from all three flavivirus subgroups.species within the flavivirus genus pose public health problems around the world. increasing cases of dengue and japanese encephalitis virus in asia, frequent outbreaks of yellow fever virus in africa and south america, and the ongoing spread of west nile virus throughout the americas, show the geographical burden of flavivirus diseases. flavivirus infections are often indistinct from and confused with other febrile illnesses. here we review the specificity of published primers, and describe a n ...200818218114
reliable detection of st. louis encephalitis virus by rt-nested pcr.st. louis encephalitis virus (slev) is a re-emerging arbovirus in south america, with reported cases in humans in argentina and brazil. this fact indicates that there is an urgent need to increase the current knowledge about this virus in order to control and prevent future cases. exhaustive epidemiological and laboratory investigation is required to ensure fast, accurate identification of the viral agent and allow prompt surveillance action by health authorities. herein, we report the developme ...200818208760
duplex reverse transcription-pcr followed by nested pcr assays for detection and identification of brazilian alphaviruses and flaviviruses.a new approach was developed for the rapid detection and identification of brazilian alphaviruses and flaviviruses. the methodology involves the genus-specific detection of alphavirus and flavivirus by a duplex reverse transcription-pcr (d-rt-pcr), followed by multiplex nested pcr (m-n-pcr) or nested pcr (n-pcr) assays for species-specific identification. by this protocol, 25 arboviruses were specifically detected and identified. detection levels between 10(1.3) and 10(3.5) 50% tissue culture in ...200515695666
epitope-blocking enzyme-linked immunosorbent assays for the detection of serum antibodies to west nile virus in multiple avian species.we report the development of epitope-blocking enzyme-linked immunosorbent assays (elisas) for the rapid detection of serum antibodies to west nile virus (wnv) in taxonomically diverse north american avian species. a panel of flavivirus-specific monoclonal antibodies (mabs) was tested in blocking assays with serum samples from wnv-infected chickens and crows. selected mabs were further tested against serum samples from birds that represented 16 species and 10 families. serum samples were collecte ...200312624027
serologic evidence of natural togavirus infections in panamanian sloths and other vertebrates.plasmas of sloths and other central panamanian wild vertebrates were tested for plaque-reduction neutralizing (prn) antibodies against four flaviviruses and one alpha-virus. forty percent of 97 two-toed sloths, choloepus hoffmanni, and 8% of 168 three-toed sloths, bradypus variegatus, were specifically positive against st. louis encephalitis (sle) virus. the prevalence of antibody against sle virus was considerably higher in sloths than in any other group of wild vertebrates tested, including bi ...19836309027
detection of the mosquito-borne flaviviruses, west nile, dengue, saint louis encephalitis, ilheus, bussuquara, and yellow fever in free-ranging black howlers (alouatta caraya) of northeastern argentina.several medically important mosquito-borne flaviviruses have been detected in argentina in recent years: dengue (denv), st. louis encephalitis (slev), west nile (wnv) and yellow fever (yfv) viruses. evidence of bussuquara virus (bsqv) and ilheus virus (ilhv) activity were found, but they have not been associated with human disease. non-human primates can act as important hosts in the natural cycle of flaviviruses and serological studies can lead to improved understanding of virus circulation dyn ...201728187130
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