Development And Preliminary Application Of DNA Chip For Identifying Nine Important Mosquitoes

Mosquitoes(Diptera:Culicidae)are of great medical importance because they could transmit many severe mosquito-borne diseases,such as malaria,dengue fever,yellow fever,Japanese encephalitis,zika virus infection and filariasis.However,the vector competence of different mosquitoes varies greatly.Therefore,the accurate and rapid identification of mosquitoes is very necessary for the prevention and control of mosquito-borne diseases.The morphological identification is usually employed in the classification of mosquitoes,but it is limited when the morphological characteristics was damaged and difficult to distinguish sibling species.Nowadays,molecular biology has been widely used in the identification of species,yet these methods are mostly focused on the identification of the complex species.Moreover,the approaches based on the PCR can't screen the massive samples each time and is not suitable for the rapid identification of mosquitoes when mosquito-borne diseases break out.Nine mosquitoes of great medical importance are used in this study including Aedes aegypti,Ae.albopictus,Armigeres subalbatus,Culex quinquefasicatus,Cx.molestus,Cx.tritaeniorhynchus,Cx.annulus,Anopheles sinensis and An.minimus.The sequences of two mitochondrial genes,NADH Dehydrogenase Subunits 2(ND2)and Cytochrome Oxidase Subunit I(COI)and a nuclear gene of Internal transcribed spacer 2(ITS2)were selected to design species-specific probes.DNA chips thus were developed to provide simultaneous identification of nine important medically species.After a series of screening and tests,this chip was applied to detect the mosquitoes in different development stages and the samples collected from the field.It was showed that the DNA chip is highly sensitive and specific,which can be employ to identify the tested mosquitoes quickly and accurately.