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The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. . https://doi.org/10.1101/291849 doi: bioRxiv preprint Mosquitoes stored for up to three weeks at the indicated temperatures were prepared by 'in-tube' crude processing. 2 µL of a 1:10 dilution of each mosquito sample was analyzed by wsp and coi LAMP-OSD assays. OSD fluorescence was imaged at endpoint using a cellphone. (A) Schematic depicting 'in-tube' method for rapid preparation of field-caught mosquitoes for LAMP-OSD analysis. (B) Comparison of coi LAMP-OSD results with morphological identification using 2X2 contingency table. (C) Tabulation of coi and wsp LAMP-OSD readout for each mosquito in the study. Ae. aegypti, Ae. albopictus and Ochlerotatus species are highlighted in blue, orange and white, respectively. 'Y' indicates a positive LAMP-OSD signal (bright fluorescence). 'N' indicates absence of any fluorescence signal in the LAMP-OSD reaction. Red boxes highlight LAMP-OSD tests that generated false positive or false negative outcomes.