RNA sequences for Aedes species, Dengue, and Chikungunya viruses

There are arthropod-borne disease outbreaks as a result of pathogen influx including arboviruses which are transmitted by strains of Aedes species that occur periodically in varying spots on the globe. The aim of this study was to determine phylogenetic relationship of Aedes mosquitoes, Dengue, and Chikungunya viruses along the Coastline of Kenya based on sequences of:

mitochondria nicotinamide adenine dehydrogenase sub unit 4 gene for Aedes species.
non-structural protein 5 gene for Dengue virus
non-structural protein 1 gene for Chikungunya virus
, Sample collection: Indoor and outdoor sampling of adults Aedes mosquitoes was done using Biogent Sentinel trap baited with solid carbon dioxide and Prokopack aspiration technique.
RNA extraction and cDNA synthesis
Pools of Aedes mosquitoes (20 mosquitoes per pool) were homogenized by a mortar and pestle and RNA extracted using 1 ml of Trizol® as earlier described. Only the unfed and gravid mosquitoes were utilised in this analysis. The female blood-fed were excluded to avoid contamination of the virus which could be in the blood meal. cDNA synthesis was done on 10 μl of the extracted RNA to generate cDNA by using EcoDry Premix (Random hexamers) (Clontech laboratories, Inc., Mountain view, CA, USA) in a procedure described earlier.
Molecular Identification and sequencing of Aedes mosquitoes and Viruses 
The mtNAD4 gene was utilised for identification of Aedes mosquitoes using primers listed (Table 1) as earlier described. Identification of serotypes of Dengue virus in the Aedes mosquit...,