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1.
Parasit Vectors ; 13(1): 468, 2020 Sep 14.
Article in English | MEDLINE | ID: mdl-32928267

ABSTRACT

BACKGROUND: Dengue viruses (DENV) can be transmitted from an adult female Aedes aegypti mosquito through the germ line to the progeny; however, there is uncertainty if this occurs at a frequency that is epidemiologically significant. We measured vertical transmission of DENV from field-reared Ae. aegypti to their F1 progeny after feeding upon blood from dengue patients. We also examined the transmission potential of F1 females. METHODS: We examined the frequency of vertical transmission in field-reared mosquitoes, who fed upon blood from acutely viremic dengue patients, and the capacity for vertically infected females to subsequently transmit virus horizontally, in two sets of experiments: (i) compared vertical transmission frequency of field-reared Ae. aegypti and Ae. albopictus, in individual progeny; and (ii) in pooled progeny derived from field- and laboratory-reared Ae. aegypti. RESULTS: Of 41 DENV-infected and isofemaled females who laid eggs, only a single female (2.43%) transmitted virus to one of the F1 progeny, but this F1 female did not have detectable virus in the saliva when 14 days-old. We complemented this initial study by testing for vertical transmission in another 460 field-reared females and > 900 laboratory-reared counterparts but failed to provide any further evidence of vertical virus transmission. CONCLUSIONS: In summary, these results using field-reared mosquitoes and viremic blood from dengue cases suggest that vertical transmission is uncommon. Field-based studies that build on these observations are needed to better define the contribution of vertical DENV transmission to dengue epidemiology.


Subject(s)
Aedes/virology , Dengue Virus/physiology , Dengue/transmission , Ovum/virology , Adolescent , Adult , Aedes/physiology , Animals , Blood/virology , Dengue/blood , Dengue/virology , Dengue Virus/genetics , Female , Humans , Infectious Disease Transmission, Vertical , Male , Pedigree , Saliva/virology , Vietnam , Young Adult
2.
Am J Trop Med Hyg ; 98(2): 402-409, 2018 02.
Article in English | MEDLINE | ID: mdl-29313471

ABSTRACT

Vietnam is endemic for dengue. We conducted a series of retrospective and prospective studies to characterize the epidemiology of dengue and population mobility patterns in Nha Trang city, Vietnam, with a view to rational design of trials of community-level interventions. A 10-year time series of dengue case notifications showed pronounced interannual variability, as well as spatial heterogeneity in ward-level dengue incidence (median annual coefficient of variation k = 0.47). Of 451 children aged 1-10 years enrolled in a cross-sectional serosurvey, almost one-third had evidence of a past dengue virus (DENV) infection, with older children more likely to have a multitypic response indicative of past exposure to ≥ 1 serotype. All four DENV serotypes were detected in hospitalized patients during 8 months of sampling in 2015. Mobility data collected from 1,000 children and young adults via prospective travel diaries showed that, although all ages spent approximately half of their daytime hours (5:00 am-9:00 pm) at home, younger age groups (≤ 14 years) spent a significantly greater proportion of their time within 500 m of home than older respondents. Together these findings inform the rational design of future trials of dengue preventive interventions in this setting by identifying 1) children < 7 years as an optimal target group for a flavivirus-naive serological cohort, 2) children and young adults as the predominant patient population for a study with a clinical end point of symptomatic dengue, and 3) substantial spatial and temporal variations in DENV transmission, with a consequent requirement for a trial to be large enough and of long enough duration to overcome this heterogeneity.


Subject(s)
Dengue/virology , Serology/methods , Adolescent , Adult , Arboviruses/drug effects , Arboviruses/pathogenicity , Child , Child, Preschool , Cluster Analysis , Cross-Sectional Studies , Dengue/epidemiology , Dengue Virus/pathogenicity , Female , Humans , Incidence , Infant , Male , Polymerase Chain Reaction/methods , Population Surveillance/methods , Serology/statistics & numerical data , Statistics, Nonparametric , Surveys and Questionnaires , Vietnam/epidemiology
3.
J Infect Dis ; 213(6): 975-84, 2016 Mar 15.
Article in English | MEDLINE | ID: mdl-26582957

ABSTRACT

The 4 dengue virus serotypes (DENV-1-4) cause the most prevalent mosquito-borne viral disease of humans worldwide. DENV-2 Asian 1 (A1) genotype viruses replaced the Asian-American (AA) genotype in Vietnam and Cambodia, after which A1 viruses containing Q or M at envelope (E) residue 160 became more prevalent than those with residue 160K in both countries (2008-2011). We investigated whether these substitutions conferred a fitness advantage by measuring neutralizing antibody titer against reporter virus particles (RVPs) representing AA, A1-160K, A1-160Q, and A1-160M, using patient sera from Vietnam and a well-characterized Nicaraguan cohort. Surprisingly, we found that A1-160Q and A1-160M RVPs were better neutralized by heterologous antisera than A1-160K. Despite this, Vietnamese patients infected with A1-160Q or A1-160M viruses had higher viremia levels than those infected with A1-160K. We thus found that independent lineages in Vietnam and Cambodia acquired a substitution in E that significantly increased polyclonal neutralization but nonetheless were successful in disseminating and infecting human hosts.


Subject(s)
Antibodies, Neutralizing/immunology , Antibodies, Viral/immunology , Dengue Virus/classification , Dengue Virus/genetics , Gene Expression Regulation, Viral/physiology , Viral Envelope Proteins/metabolism , Amino Acid Substitution , Biological Evolution , Cloning, Molecular , Dengue/blood , Dengue/immunology , Dengue/virology , Evolution, Molecular , Humans , Phylogeny , Serotyping , Viral Envelope Proteins/genetics
4.
PLoS Negl Trop Dis ; 9(9): e0004052, 2015.
Article in English | MEDLINE | ID: mdl-26325059

ABSTRACT

Dengue virus (DENV) infection of an individual human or mosquito host produces a dynamic population of closely-related sequences. This intra-host genetic diversity is thought to offer an advantage for arboviruses to adapt as they cycle between two very different host species, but it remains poorly characterized. To track changes in viral intra-host genetic diversity during horizontal transmission, we infected Aedes aegypti mosquitoes by allowing them to feed on DENV2-infected patients. We then performed whole-genome deep-sequencing of human- and matched mosquito-derived DENV samples on the Illumina platform and used a sensitive variant-caller to detect single nucleotide variants (SNVs) within each sample. >90% of SNVs were lost upon transition from human to mosquito, as well as from mosquito abdomen to salivary glands. Levels of viral diversity were maintained, however, by the regeneration of new SNVs at each stage of transmission. We further show that SNVs maintained across transmission stages were transmitted as a unit of two at maximum, suggesting the presence of numerous variant genomes carrying only one or two SNVs each. We also present evidence for differences in selection pressures between human and mosquito hosts, particularly on the structural and NS1 genes. This analysis provides insights into how population drops during transmission shape RNA virus genetic diversity, has direct implications for virus evolution, and illustrates the value of high-coverage, whole-genome next-generation sequencing for understanding viral intra-host genetic diversity.


Subject(s)
Aedes/virology , Dengue Virus/classification , Dengue Virus/genetics , Dengue/virology , Genetic Variation , Adult , Animals , Dengue Virus/isolation & purification , Female , Gastrointestinal Tract/virology , High-Throughput Nucleotide Sequencing , Humans , Male , Polymorphism, Single Nucleotide , RNA, Viral/genetics , Salivary Glands/virology , Selection, Genetic , Viral Nonstructural Proteins/genetics , Young Adult
5.
J Virol Methods ; 177(2): 168-73, 2011 Nov.
Article in English | MEDLINE | ID: mdl-21843553

ABSTRACT

Dengue is mosquito-borne virus infection that annually causes ~50 million clinically apparent cases worldwide. An internally controlled one-step real-time multiplex RT-PCR assay was developed for detection and quantitation of DENV RNA in plasma sample by using specific primers and fluorogenic TaqMan probes. All primers and probes targeted sequences near the 3' end of the NS5 gene. The method comprised two multiplex assays and was validated for sensitivity, specificity, linearity, reproducibility and precision. An internal control template was spiked into each clinical specimen to provide quality assurance for each experimental step. The assay allowed for detection of between 0.5 and 3 infectious particles per mL, is rapid and has been operationally characterized in 287 Vietnamese dengue patients from two therapeutic intervention trials at the Hospital for Tropical Diseases, Ho Chi Minh City, Viet Nam.


Subject(s)
Dengue Virus/genetics , Dengue/diagnosis , Multiplex Polymerase Chain Reaction/methods , RNA, Viral/blood , Reverse Transcriptase Polymerase Chain Reaction/methods , Dengue/virology , Equartevirus/genetics , Humans , Limit of Detection , Microbiological Techniques , Plasmids/genetics , Plasmids/metabolism , Reproducibility of Results , Sensitivity and Specificity , Viral Nonstructural Proteins/genetics
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