A Simple Technique for the Detection of Dengue Antigen in Mosquitoes by Immunofluorescence

Kuberski, Timothy T.; Rosen, Léon

doi:10.4269/ajtmh.1977.26.533

Cited by 135 publications

(75 citation statements)

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“…Virus detection in the mosquito, regardless of the species, is generally performed by the direct fluorescent-antibody DFA test on mosquito tissues, usually brain or salivary glands (47,50,86). The direct conjugate is prepared from pooled human serum and has broadly reactive anti-dengue (or anti-flavivirus) activity.…”

Section: Virus Isolationmentioning

confidence: 99%

Dengue and Dengue Hemorrhagic Fever

Gubler¹

1997

Tropical Infectious Diseases

633

756

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Section: Virus Isolationmentioning

confidence: 99%

Dengue and Dengue Hemorrhagic Fever

Gubler¹

1997

Tropical Infectious Diseases

633

756

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“…DENV Antigen Detection in Mosquito Tissue. DENV antigen was detected in mosquito tissue by indirect IFA (39). Dissected midguts and salivary glands were fixed in 4% (vol͞vol) paraformaldehyde in PBS.…”

Section: Denv-2 and -4 Challenge Experimentsmentioning

confidence: 99%

Engineering RNA interference-based resistance to dengue virus type 2 in genetically modified Aedes aegypti

Franz

Sánchez-Vargas

Adelman

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

370

375

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Mosquitoes (Aedes aegypti) were genetically modified to exhibit impaired vector competence for dengue type 2 viruses (DENV-2). We exploited the natural antiviral RNA interference (RNAi) pathway in the mosquito midgut by constructing an effector gene that expresses an inverted-repeat (IR) RNA derived from the premembrane protein coding region of the DENV-2 RNA genome. The A. aegypti carboxypeptidase A promoter was used to express the IR RNA in midgut epithelial cells after ingestion of a bloodmeal. The promoter and effector gene were inserted into the genome of a white-eye Puerto Rico Rexville D (Higgs' white eye) strain by using the nonautonomous mariner MosI transformation system. A transgenic family, Carb77, expressed IR RNA in the midgut after a bloodmeal. Carb77 mosquitoes ingesting an artificial bloodmeal containing DENV-2 exhibited marked reduction of viral envelope antigen in midguts and salivary glands after infection. DENV-2 titration of individual mosquitoes showed that most Carb77 mosquitoes poorly supported virus replication. Transmission in vitro of virus from the Carb77 line was significantly diminished when compared to control mosquitoes. The presence of DENV-2-derived siRNAs in RNA extracts from midguts of Carb77 and the loss of the resistance phenotype when the RNAi pathway was interrupted proved that DENV-2 resistance was caused by a RNAi response. Engineering of transgenic A. aegypti that show a high level of resistance against DENV-2 provides a powerful tool for developing population replacement strategies to control transmission of dengue viruses.RNA silencing ͉ transgenesis ͉ genetic control ͉ mosquito ͉ dengue disease D engue viruses (DENV) [Flaviviridae; Flavivirus; DENV types 1-4 (DENV-1-4)] threaten public health in Ͼ100 countries and infect an estimated 50 million people annually (1, 2). The mosquito, Aedes aegypti, is the principal vector for epidemic dengue disease (3). The urban DENV transmission cycle involves only humans and mosquitoes. No DENV vaccines are currently available, and vector control strategies that minimize human-mosquito contact have largely failed (4, 5). New control strategies are needed. One possible strategy is to replace vector populations competent to transmit DENVs with pathogenincompetent vectors (6). The essential features of this genetic control strategy are to identify genes that express antiviral molecules in the vector, link this gene (or genes) to a genetic drive system [transposable elements (TE), meiotic drive, or homing endonuclease genes] and introgress the gene(s) into field populations (7-9). A key step in developing this control strategy is to identify effector genes that, when expressed in the vector, inhibit DENV replication. Proof of principle for RNA interference (RNAi)-like disruption of DENV-2 vector competence was previously demonstrated by using a nonheritable alphavirus expression system (10). Applying the principle of heritable gene silencing in transgenic Drosophila melanogaster, Caenorhabditis elegans, and A. aegypti (11-13), we gene...

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“…includes isolation and propagation of viruses, reverse transcriptasepolymerase chain reaction (RT-PCR) assays, or enzymelinked immunosorbent assay or immunofluorescence assay (Kuberski et al 1977, Chan et al 1994, Sithiprasasna et al 1994, Rohani et al 1997, Chow et al 1998, Samuel and Tiyagi 2006. However, the need for expensive specialized laboratories and equipment and highly trained staff has made these techniques impractical for regular surveillance of DENVinfected mosquitoes.…”

mentioning

confidence: 99%

Evaluation of the Dengue NS1 Ag Strip^®for Detection of Dengue Virus Antigen inAedes aegypti(Diptera: Culicidae)

Tan

Wong

et al. 2011

Vector-Borne and Zoonotic Diseases

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Dengue fever is currently one of the most important mosquito-borne diseases that affect humans. With neither vaccines nor treatment available, prevention of the disease relies heavily on surveillance and control of mosquito vectors. In the present study, we have evaluated and showed the potential use of the Dengue NS1 Ag Strip Ò for the detection of dengue virus (DENV) in Aedes aegypti. Initial results showed that the sensitivity of the test kit in detecting DENV in wild-caught mosquitoes is comparable to that of real-time reverse transcriptase-polymerase chain reaction. The detection of naturally infected Ae. aegypti with the NS1 rapid test kit in our dengue cluster investigation further illustrates its potential use for surveillance of DENV in wild mosquito populations. The kit can easily be used in a simple field station, and minimal training is required. The results can be obtained in less than an hour. Employment of the kit in the field could help guide mosquito control operations in the prioritization of resources in controlling the transmission of DENV. In this study the potential of the kit for field surveillance of infected dengue vectors, which are epidemiologically important, has been demonstrated.

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A Simple Technique for the Detection of Dengue Antigen in Mosquitoes by Immunofluorescence

Cited by 135 publications

References 0 publications

Dengue and Dengue Hemorrhagic Fever

Dengue and Dengue Hemorrhagic Fever

Engineering RNA interference-based resistance to dengue virus type 2 in genetically modified Aedes aegypti

Evaluation of the Dengue NS1 Ag Strip^®for Detection of Dengue Virus Antigen inAedes aegypti(Diptera: Culicidae)

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A Simple Technique for the Detection of Dengue Antigen in Mosquitoes by Immunofluorescence

Cited by 135 publications

References 0 publications

Dengue and Dengue Hemorrhagic Fever

Dengue and Dengue Hemorrhagic Fever

Engineering RNA interference-based resistance to dengue virus type 2 in genetically modified Aedes aegypti

Evaluation of the Dengue NS1 Ag Strip®for Detection of Dengue Virus Antigen inAedes aegypti(Diptera: Culicidae)

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Evaluation of the Dengue NS1 Ag Strip^®for Detection of Dengue Virus Antigen inAedes aegypti(Diptera: Culicidae)