Additional file 2 of Impact of temperature on vector competence of Culex pipiens molestus: implications for Usutu virus transmission in temperate regions

Additional file 2. Figure A2: Proportion of body positive and saliva positive samples at 22 ˚C, 20 ˚C and 18 ˚C. Samples incubated at 18 ˚C were analysed using an elution buffer at half the volume of samples at 20 ˚C. Pools of mosquitoes were fed spiked blood containing USUV at a titre of 4 × 107 PFU/ml and were tested by real-time RT-PCR. Saliva samples at 0 dpi were taken immediately after blood feeding and residual virus present in mouthparts likely contaminated saliva as it was expectorated giving rise to false-positive results (A). Here, the proportion of saliva positive samples is forced to one. Body samples at 0 dpi contained virus in the blood meal and so the proportion of body positive samples was 1 at 0 dpi (B).