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. 2014 Apr 1:7:155.
doi: 10.1186/1756-3305-7-155.

Preliminary evaluation on the efficiency of the kit Platelia Dengue NS1 Ag-ELISA to detect dengue virus in dried Aedes aegypti: a potential tool to improve dengue surveillance

Gabriel Sylvestre, Mariana Gandini, Josélio M G de Araújo, Claire F Kubelka, Ricardo Lourenço-de-Oliveira, Rafael Maciel-de-Freitas  1
Affiliations

Preliminary evaluation on the efficiency of the kit Platelia Dengue NS1 Ag-ELISA to detect dengue virus in dried Aedes aegypti: a potential tool to improve dengue surveillance

Gabriel Sylvestre et al. Parasit Vectors. .

Abstract

Background: Surveillance is a critical component of any dengue prevention and control programme. Herein, we investigate the efficiency of the commercial kit Platelia Dengue NS1 Ag-ELISA to detect dengue virus (DENV) antigens in Aedes aegypti mosquitoes infected under laboratory conditions.

Methods: Under insectary conditions, four to five day-old mosquitoes were orally challenged with DENV-2 titer of 3.6 x 10⁵ PFU equivalent/ml, incubated for 14 days and then killed. At ten time-points following mosquito death (0, 6, 12, 24, 72, 96, 120, 144 and 168 h), i.e., during a one-week period, dried mosquitoes were comparatively tested for the detection of the NS1 antigen with other methods of detection, such as qRT-PCR and virus isolation in C6/36 cells.

Results: We first observed that the NS1 antigen was more effective in detecting DENV-2 in Ae. aegypti between 12 and 72 h after mosquito death when compared with qRT-PCR. A second round involved comparing the sensitivity of detection of the NS1 antigen and virus isolation in C6/36 cells. The NS1 antigen was also more effective than virus isolation, detecting DENV-2 at all time-points, i.e., up to 168 h after mosquito death. Meanwhile, virus isolation was successful up to 96 h after Ae. aegypti death, but the number of positive samples per time period presented a tendency to decline progressively over time. From the 43 samples positive by the virus isolation technique, 38 (88.4%) were also positive by the NS1 test.

Conclusion: Taken together, these results are the first to indicate that the NS1 antigen might be an interesting complementary tool to improve dengue surveillance through DENV detection in dried Ae. aegypti females.

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Figures

Figure 1
Figure 1
Comparison between the median optical density revealed by the Platelia Dengue NS1 Ag-ELISA (bars) and the median number of RNA copies determined by qRT-PCR (line) of Aedes aegypti mosquitoes killed naturally and analyzed for dengue virus detection between 0–72 h after death.
See this image and copyright information in PMC

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