Direct nucleic acid analysis of mosquitoes for high fidelity species identification and detection of Wolbachia using a cellphone

Sanchita Bhadra; Timothy E. Riedel; Miguel A. Saldaña; Shivanand Hegde; Nicole Pederson; Grant L. Hughes; Andrew D. Ellington

The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. and wAlbB Wolbachia-infected Ae. albopictus mosquitoes by coi and wsp LAMP-OSD assays, respectively. 2 µL of mosquito samples were either added directly to LAMP-OSD assays or subjected to 10-fold serial dilution in water prior to introduction in LAMP-OSD reactions. LAMP-OSD reactions without any macerated mosquito samples served as negative controls. Assays lacking LAMP primers but containing macerated mosquito samples served as controls for monitoring background fluorescence. (C) Specificity analysis of coi and wsp LAMP-OSD assays. The coi LAMP-OSD assay was challenged with 2 µL of Ae. albopictus in-syringe prepared crude mosquito sample. Similarly, the wsp LAMP-OSD assay was challenged with Ae. aegypti sample. In all experiments, OSD fluorescence was imaged at endpoint using a cellphone. Individual normal or blood-fed Ae. aegypti and Ae. albopictus mosquitoes were prepared by the in-syringe method. 2 µL of these mosquito samples were analyzed by coi and wsp LAMP-OSD assays. LAMP-OSD reactions without any macerated mosquito samples served as negative controls. Assays lacking LAMP primers but containing macerated mosquito samples served as controls for monitoring background fluorescence. LAMP-OSD fluorescence was imaged at endpoint using a cellphone.