Abstract
BackgroundSurveillance 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.MethodsUnder 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.ResultsWe 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.ConclusionTaken 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.
Highlights
Surveillance is a critical component of any dengue prevention and control programme
Dengue serodiagnosis is usually based on anti-dengue virus (DENV) IgM and IgG detection measured by MAC-ELISA and IgG-ELISA
Distinct lots of 60 Ae. aegypti females previously fed with a blood meal containing DENV-2 were examined respectively by NS1 Ag-ELISA and quantitative reverse transcriptase polymerase chain reaction (qRT-PCR)
Summary
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. Dengue viruses belong to the Flaviviridae family as four antigenically related but distinct serotypes designated DENV1, −2, −3 and −4. Worldwide spread vector borne disease with higher incidence in the tropics due to the distribution of its vector, the Aedes aegypti mosquito. Dengue serodiagnosis is usually based on anti-DENV IgM and IgG detection measured by MAC-ELISA and IgG-ELISA. Specific antibody detection has a limitation concerning the acute phase of the disease, since the detection of DENV IgM takes 3 to 5 days and anti-DENV IgG requires 1 to 14 days, depending upon primary or secondary dengue disease [7]
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