In response to the report of the fatal BRBV case from eastern Kansas in 2014, we retrospectively tested ticks for BRBV that were collected during spring and summer 2013 from 6 sites in northwestern Missouri, [approximately equal to]240 km from Bourbon County (Figure 1).
Sequence and reporter information for BRBV primer/probe sets nucleoprotein (NP) 1 and polymerase basic (PB) 1, which we used for virus detection and confirmation, respectively, are as previously described (4).
americanum colony with serial dilutions of BRBV, original strain (1,4).
We extracted and purified BRBV RNA from HuH-7 V1 harvest from 2 pools that were rRT-PCR positive as previously described (1,4).
Reextraction and testing of the original tick homogenates using both primer/probe sets NP1 and PB1 confirmed BRBV RNA in all 3 pools.
To confirm the molecular identification of BRBV, we selected 2 pools, MO-2013-1246 of male adults and MO-2013-2499 of nymphs, for high-throughput sequencing and phylogenetic analysis.