Publications

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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
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
saint louis encephalitis virus, brazil. 200717370543
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
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
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
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
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
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
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
phylogenetic analysis of brazilian flavivirus using nucleotide sequences of parts of ns5 gene and 3' non-coding regions.viruses of the genus flavivirus, which are arboviruses, of the flaviviridae family, are amongst the most important agents of infectious disease in brazil, causing human infections with a high morbility and mortality. in this work, the phylogeny of 14 virus amplicon sequences that were obtained by rt-pcr with universal primers for mosquito-borne flavivirus were studied. the amplicons included a region of the flavivirus genome of 129 nucleotides at the 3' terminus of the ns5 gene and the 145 initi ...200111311426
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
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
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
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
recovery of west nile virus envelope protein domain iii chimeras with altered antigenicity and mouse virulence.flaviviruses are positive-sense, single-stranded rna viruses responsible for millions of human infections annually. the envelope (e) protein of flaviviruses comprises three structural domains, of which domain iii (eiii) represents a discrete subunit. the eiii gene sequence typically encodes epitopes recognized by virus-specific, potently neutralizing antibodies, and eiii is believed to play a major role in receptor binding. in order to assess potential interactions between eiii and the remainder ...201626912625
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
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
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