Volume 27, Number 4—April 2021
Venezuelan Equine Encephalitis Complex Alphavirus in Bats, French Guiana
|Name||Sequence, 5′ → 3′||Concentration|
|Forward primer||CATTGTCATAGCCAGCAGAGTTCT||400 nM|
|Reverse primer||GACTTGATACCTTTGACGATGTTGTC||400 nM|
|Probe (FAM-labeled)||CGCGAACGTCTGACCAACTCACCCT||200 nM|
*We carried out 25 μL real-time RT-PCR reactions using the Superscript III one-step RT-PCR system with Platinum Taq polymerase (Thermo Fisher Scientific, https://www.thermofisher.com). Reactions were set up with 5 μL extracted RNA; 12.5 μL of 2× reaction buffer; 0.4 μL of a 50 mM magnesium sulfate solution; 1 μg of nonacetylated bovine serum albumin; and 1 μL enzyme. Amplification was conducted at 50°C for 15 min, followed by 95°C for 3 min and 45 cycles of 95°C for 15 s and 58°C for 30 s with fluorescence, read at the 58° annealing/extension step on a LightCycler 480 thermocycler (Roche, https://www.roche.com). FAM, fluorescein amidite; RT-PCR, reverse transcription PCR.
1These authors contributed equally to this article.