P12 - Effect of Mosquito Age and Specimen Storage Duration on Aedes aegypti RNA-based Assays

Abstract: We evaluated the effect of storage duration of Aedes aegypti on the ability to extract viable RNA for molecular age grading. Colony Ae. aegypti (Rockefeller strain) adults reared in incubators to 4 d, 10 d, 21 d, and 27 d were killed and stored at 28°C for 1 h, 2 h, 5 h, 8 h, 24 h before RNA extraction. Reverse transcription-PCR was used to detect three genetic markers of age and one control gene. The Cq values from the PCR and the RNA Integrity Numbers were used to determine the quality of RNA from specimens at various ages and storage durations. Thus far, initial outcomes demonstrate substantial RNA degradation at 8 h and 24 h timepoints for all ages tested with an inability to detect age related genes. These results will be used to validate molecular age grading methodologies for Ae. aegypti and inform collection and storage of field specimens destined for molecular assays.