Visually read LAMP-OSD analysis of crudely processed individual mosquitoes.

(A) Schematic depicting rapid preparation of crude mosquito samples for direct LAMP-OSD analysis by performing in-syringe individual mosquito maceration. (B) Analysis of individual Ae. aegypti 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.