Evolution of Dengue Virus Type 3 Genotype III in Venezuela: Diversification, Rates and Population Dynamics

<p>Abstract</p> <p>Background</p> <p>Dengue virus (DENV) is a member of the genus <it>Flavivirus </it>of the family <it>Flaviviridae</it>. DENV are comprised of four distinct serotypes (DENV-1 through DENV-4) and each serotype can be divided in different genotypes. Currently, there is a dramatic emergence of DENV-3 genotype III in Latin America. Nevertheless, we still have an incomplete understanding of the evolutionary forces underlying the evolution of this genotype in this region of the world. In order to gain insight into the degree of genetic variability, rates and patterns of evolution of this genotype in Venezuela and the South American region, phylogenetic analysis, based on a large number (<it>n </it>= 119) of envelope gene sequences from DENV-3 genotype III strains isolated in Venezuela from 2001 to 2008, were performed.</p> <p>Results</p> <p>Phylogenetic analysis revealed an <it>in situ </it>evolution of DENV-3 genotype III following its introduction in the Latin American region, where three different genetic clusters (A to C) can be observed among the DENV-3 genotype III strains circulating in this region. Bayesian coalescent inference analyses revealed an evolutionary rate of 8.48 × 10^-4substitutions/site/year (s/s/y) for strains of cluster A, composed entirely of strains isolated in Venezuela. Amino acid substitution at position 329 of domain III of the E protein (A→V) was found in almost all E proteins from Cluster A strains.</p> <p>Conclusions</p> <p>A significant evolutionary change between DENV-3 genotype III strains that circulated in the initial years of the introduction in the continent and strains isolated in the Latin American region in recent years was observed. The presence of DENV-3 genotype III strains belonging to different clusters was observed in Venezuela, revealing several introduction events into this country. The evolutionary rate found for Cluster A strains circulating in Venezuela is similar to the others previously established for this genotype in other regions of the world. This suggests a lack of correlation among DENV genotype III substitution rate and ecological pattern of virus spread.</p

West Nile Virus, Venezuela

Submitted by Sandra Infurna ([email protected]) on 2020-04-01T17:32:07Z No. of bitstreams: 1 AnthonyE_Guimaraes_etal_IOC_2007.pdf: 108341 bytes, checksum: db1489aeeb51b9f823abd6e1273378ef (MD5)Approved for entry into archive by Sandra Infurna ([email protected]) on 2020-04-01T17:51:00Z (GMT) No. of bitstreams: 1 AnthonyE_Guimaraes_etal_IOC_2007.pdf: 108341 bytes, checksum: db1489aeeb51b9f823abd6e1273378ef (MD5)Made available in DSpace on 2020-04-01T17:51:00Z (GMT). No. of bitstreams: 1 AnthonyE_Guimaraes_etal_IOC_2007.pdf: 108341 bytes, checksum: db1489aeeb51b9f823abd6e1273378ef (MD5) Previous issue date: 2007University of Massachusetts Medical School. Center for Infectious Disease and Vaccine Research. Worcester, MA, USA.Universidad de Carabobo Biomed. Maracay, Venezuela.Universidad Central de Venezuela. Caracas, Venezuela.Coleccion Ornitologica Phelps. Caracas, Venezuela.New York State Department of Health. Albany, New York, USA / State University of New York at Albany. Albany, New York, USA.New York State Department of Health. Albany, New York, USA / State University of New York at Albany. Albany, New York, USA.New York State Department of Health. Albany, New York, USA / State University of New York at Albany. Albany, New York, USA.Universidad Central de Venezuela. Maracay, Venezuela.Instituto Nacional de Investigaciones Agrícolas. Maracay, Venezuela.Universidad Central de Venezuela, Caracas, VenezuelaFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.Centers for Disease Control and Prevention. San Juan, Puerto Rico, USA.Universidad del Zulia. Maracaibo, Venezuela.Universidad de Carabobo Biomed. Maracay, Venezuela.Ministerio de Salud Insalud. Carabobo, Venezuela.Universidad Central de Venezuela, Caracas, VenezuelaUniversidad de Carabobo Biomed. Maracay, Venezuela.Centers for Disease Control and Prevention.Fort Collins, Colorado, USA.Harvard School of Public Health. Boston, Massachusetts, USA.New York State Department of Health. Albany, New York, USA / State University of New York at Albany. Albany, New York, USA

Multicountry Prospective Clinical Evaluation of Two Enzyme-Linked Immunosorbent Assays and Two Rapid Diagnostic Tests for Diagnosing Dengue Fever

We evaluated four dengue diagnostic devices from Alere, including the SD Bioline Dengue Duo (nonstructural [NS] 1 Ag and IgG/IgM), the Panbio Dengue Duo Cassette (IgM/IgG) rapid diagnostic tests (RDTs), and the Panbio dengue IgM and IgG capture enzyme-linked immunosorbent assays (ELISAs) in a prospective, controlled, multicenter study in Peru, Venezuela, Cambodia, and the United States, using samples from 1,021 febrile individuals. Archived, well-characterized samples from an additional 135 febrile individuals from Thailand were also used. Reference testing was performed on all samples using an algorithm involving virus isolation, in-house IgM and IgG capture ELISAs, and plaque reduction neutralization tests (PRNT) to determine the infection status of the individual. The primary endpoints were the clinical sensitivities and specificities of these devices. The SD Bioline Dengue Duo had an overall sensitivity of 87.3% (95% confidence interval [CI], 84.1 to 90.2%) and specificity of 86.8% (95% CI, 83.9 to 89.3%) during the first 14 days post-symptom onset (p.s.o.). The Panbio Dengue Duo Cassette demonstrated a sensitivity of 92.1% (87.8 to 95.2%) and specificity of 62.2% (54.5 to 69.5%) during days 4 to 14 p.s.o. The Panbio IgM capture ELISA had a sensitivity of 87.6% (82.7 to 91.4%) and specificity of 88.1% (82.2 to 92.6%) during days 4 to 14 p.s.o. Finally, the Panbio IgG capture ELISA had a sensitivity of 69.6% (62.1 to 76.4%) and a specificity of 88.4% (82.6 to 92.8%) during days 4 to 14 p.s.o. for identification of secondary dengue infections. This multicountry prospective study resulted in reliable real-world performance data that will facilitate data-driven laboratory test choices for managing patient care during dengue outbreaks

Circulating Strains of Human Respiratory Syncytial Virus in Central and South America

Human respiratory syncytial virus (HRSV) is a major cause of viral lower respiratory tract infections among infants and young children. HRSV strains vary genetically and antigenically and have been classified into two broad subgroups, A and B (HRSV-A and HRSV-B, respectively). To date, little is known about the circulating strains of HRSV in Latin America. We have evaluated the genetic diversity of 96 HRSV strains by sequencing a variable region of the G protein gene of isolates collected from 2007 to 2009 in Central and South America. Our results show the presence of the two antigenic subgroups of HRSV during this period with the majority belonging to the genotype HRSV-A2

Cloning alphavirus and flavivirus sequences for use as positive controls in molecular diagnostics

The purpose of the study was to obtain a positive control to validate molecular techniques (reverse transcription– polymerase chain reaction [RT-PCR]) used in the diagnosis and research of viral infections. From strains of Chikungunya virus (CHIKV), Zika virus, and Dengue virus (DENV-1, DENV-2, DENV- 3, and DENV-4) viral RNAs were extracted to obtain complementary DNA using RT-PCR from the nsP4 (CHIKV), NS5 (Zika virus), C/prM-M, and 5′UTR-C (DENV-1, DENV-2, DENV-3, DENV-4) sequences, which were cloned into pGEM®-T Easy. Cloning was confirmed through colony PCR, from which plasmid DNA was extracted for fragment cloning verification. Cloning of cDNA corresponding to nsP4, NS5, C/prM-M, and 5′UTR-C of the different viral agents was achieved. In conclusion, recombinant plasmids were obtained with each of the sequences specified for further assessment as positive controls in molecular techniques in an effort to avoid the use of cell cultures, which can be costly, time-consuming, and potentially dangerous

El ensayo inmunoenzimatico en microgotas sobre nitrocelulosa (Dot-ELISA) en el diagnostico de la enfermedad de Chagas: I. Estudio comparativo de dos preparaciones antigenicos de Trypanosoma cruzi The Dot-Enzyme linked immunosorbent assay (Dot-ELISA) in the diagnosis of Chagas-disease: I. Comparative study of two antigenic preparations of Trypanosoma cruzi

Se estudia el Ensayo Inmunoenzimático en Microgotas sobre Nitrocelulosa (Dot-ELISA)comparando dos preparados antigénicos de formas epimastigotas de cultivo de T. cruzi: 1) la fracción citoplasmática (antígeno citoplasmático y 2) el parásito total fijado previamente con formaldehido (antígeno integral). Se usaron sueros de: 95 pacientes chagásicos con serología convencional positiva, cardiopatía crónica y algunos con xenodiagnóstico positivo; 42 personas sanas y 32 con miocardipatía crónica con serología negativa y 74 pacientes con diferentes patologías incluyendo: sífilis, toxoplasmosis, lupus eritematoso diseminado, con factor reumatoide, leishmaniasis visceral, y leishmaniasis cutánea. Definidos los títulos diagnósticos (cut-off) de 1:512 con antígeno citoplasmático y de 1: 128 con antígeno integral, la especificidad fue 96% para el primero y de 100% para el segundo; mientras que la sensibilidad fue de 100% para ambas. En el estudio comparativo con las pruebas serológicas convencionales examinando 147 sueros tomados de personas referidas al laboratório, Dot-ELISA con antígeno citoplasmático presentó índices deco-positividad de 1,0, co-negatividad de 0,989 y eficiencia 0,993. Dot-ELIS con antígeno integral dió 1,0, 0,979 y 0,986 respectivamente. De acuerdo con esta evaluación, la técnica Dot-ELISA con antígeno integral se presenta como una alternativa práctica para el diagnóstico serológico de la enfermedad de Chagas.<br>Using the Dot-ELISA technique, two antigenic preparations of Trypanosoma cruzi epimastigote forms have been compared for the diagnosis of Chagas' disease: (1) The citoplasmic fraction (citoplasmic antigen) and (2) whole fixed epimastigotes (integral antigen). There was been used sera from 95 chagasic patients with chronic cadiomyopathy, positive conventional serology and either positive or negative xenodiagnosis; 74 subjects with negative conventional serology, and either clinically normal or presenting cardiomyopathy; 74 patients with different diseases including siphilis,toxoplasmosis, leisjmaniasis or autoantibodies such as rheumatoid factor and antinuclear antibodies. By defining the diagnostic titers (cut Off): 1:512 for citoplasmatic antigen and 1:128 for the integral antigen, a sensitivity of 100% has been obtained with both antigenic preparations, being the specificity of 96% for the former and 100% for the latter when leishmaniasis sera were not included. A comparative study with conventional serology was carried out using 147 sera from a Laboratory of Chagas' diagnosis; Dot-ELISA with citoplasmic antigen showed co-positivity index of 1.0, co-negativity 0.989 and efficiency of 0.993, and Dot-ELISA with integral antigen 1.0, 0.979 and 0.986 respectively. According to this evaluation, Dot-ELISA using whole formalin fixed epimastigotes might be a practical alternative for the serological diagnosis of Chagas' disease

Dinamic changes of pro-inflammatory cytokines, adhesion molecules and lymphocytes activation markers as early indicators of diseases severity in patients with Dengue

Several immunopathogenic mechanisms have been proposed to explain the massive increase of vascular permeability observed in the severe forms of infection by Dengue Virus (DENV). Our aim was to determine the kinetic changes of inflammatory mediators (IL-8, TNF- α), soluble early lymphocyte activation markers (sIL-2R, sTNF-Rp75) and soluble fractions of cell adhesion molecules (sICAM-1 and sVCAM-1) as indicators for early recognition of disease severity in patients with laboratory-confirmed dengue. Twenty patients classified as Dengue±Warning Signs (D±WS) and thirty patients with Severe Dengue (SD) were included in the study. Serums of apparently healthy individuals were included as controls. Compared with normal subjects, D±WS cases did not show significant differences in the levels of IL-8 or TNF-α during the acute nor in the critical stages of the disease; however, in D±WS cases levels of sICAM-1 and sVCAM-1 were higher than controls during both phases; in contrast, significant increase of sTNF-p75 and sIL2R levels were observed during the critical phase of the disease. Compared with both dengue patients and controls, patients with SD showed significant rise in the levels of IL-8 and TNF-α during the critical phase of the disease and a significant increase in adhesion molecules were detected in both phases, but the highest levels of sVCAM-1 and sIL-2R were observed only during the acute stage of the disease. In conclusion, sIL-2R and sVCAM-1, as early markers of lymphocyte and endothelial activation, would serves as indicators of severity during the acute phase of dengue infection

Cambios dinámicos de citoquinas proinflamatorias, moléculas de adhesión y marcadores de activación linfocítica como indicadores tempranos de severidad en pacientes con Dengue

Varios mecanismos inmuno-patogénicos se han propuesto para explicar el incremento masivo de la permeabilidad vascularobservada en las formas severas de la infección por el Virus del Dengue (DENV). El objetivo del estudio fue determinar loscambios cinéticos de mediadores inflamatorios (IL-8, TNF-α), marcadores soluble de activación linfocítica temprana (sIL-2R,sTNF-Rp75) y fracciones solubles de moléculas de adhesión celular (sICAM-1, sVCAM-1) como marcadores tempranos deseveridad en pacientes con dengue. Veinte pacientes clasificados como Dengue (Dengue±Signos de Alarma,D±WS) y treintapacientes con Dengue Severo (DS) fueron incluidos en el estudio. Suero de individuos aparentemente saludables fueronincluidos como controles. En comparación con los individuos controles, los casos con Dengue mostraron niveles de IL-8 yTNF con diferencias no significativas en la fase febril o crítica de la enfermedad; sin embargo, un incremento significativo desICAM-1 y sVCAM-1 ocurrió en ambas fases, mientras que los niveles de sIL2R y sTNF-p75 se elevaron significativamentesolo en la fase crítica de la enfermedad. En comparación con los casos con dengue y controles, los pacientes con DS mostraron diferencias significativas en los niveles de IL-8 y TNF-α durante la fase crítica y un incremento significativo demoléculas de adhesión en ambas fases, pero los niveles más elevados de sVCAM-1 y sIL-2R fueron observados en la fasefebril. En conclusión, sIL-2R y sVCAM-1, como marcadores tempranos de activación linfocítica y endotelial, servirían comoindicadores de severidad en la fase aguda de la infección por el virus del dengue.Dinamic changes of pro-inflammatory cytokines, adhesion molecules and lymphocytesactivation markers as early indicators of diseases severity in patients with DengueAbstractSeveral immunopathogenic mechanisms have been proposed to explain the massive increase of vascular permeabilityobserved in the severe forms of infection by Dengue Virus (DENV). Our aim was to determine the kinetic changes ofinflammatory mediators (IL-8, TNF- α), soluble early lymphocyte activation markers (sIL-2R, sTNF-Rp75) and solublefractions of cell adhesion molecules (sICAM-1 and sVCAM-1) as indicators for early recognition of disease severity inpatients with laboratory-confirmed dengue. Twenty patients classified as Dengue±Warning Signs (D±WS) and thirty patientswith Severe Dengue (SD) were included in the study. Serums of apparently healthy individuals were included as controls.Compared with normal subjects, D±WS cases did not show significant differences in the levels of IL-8 or TNF-α during theacute nor in the critical stages of the disease; however, in D±WS cases levels of sICAM-1 and sVCAM-1 were higher thancontrols during both phases; in contrast, significant increase of sTNF-p75 and sIL2R levels were observed during the criticalphase of the disease. Compared with both dengue patients and controls, patients with SD showed significant rise in thelevels of IL-8 and TNF-α during the critical phase of the disease and a significant increase in adhesion molecules weredetected in both phases, but the highest levels of sVCAM-1 and sIL-2R were observed only during the acute stage of thedisease. In conclusion, sIL-2R and sVCAM-1, as early markers of lymphocyte and endothelial activation, would serves asindicators of severity during the acute phase of dengue infection