DNA from ticks that screened positive for SFGR were subjected to semi-nested PCR amplification of rompA using primers Rr190-70, Rr190-701, and Rr190-602 and the intergenic region (IGR) using primary and nested primers RR0155-rpm B (Eremeeva et al., 2006; Shapiro et al., 2010; Wikswo et al., 2008). Briefly, 20 μL of 2X Taq Master Mix (Qiagen, Valencia, CA), 2 μL of forward primer Rr190-70 (20 mM), 2 μL of reverse primer Rr190-701/Rr190-602 (20 mM), 14 μL of nuclease-free H2O, and 2 μL of DNA was amplified using PCR cycling conditions of 95 °C for 3 min followed by 35 cycles of 95 °C for 20 s, 57 °C for 30 s, and 68 °C for 2 min and then 72 °C for 5 min before holding the products at 4 °C. For the IGR PCR amplification, 20 μL of 2X Taq Master Mix (Qiagen, Valencia, CA, USA), 1 μL of forward primer RR 0155 PF (20 mM), 1 μL of reverse primer 0155 PR (20 mM), 16 μL of nuclease-free H2O, and 2 μL of DNA was amplified using PCR cycling conditions of 95 °C for 5 min followed by 35 cycles of 95 °C for 30 s, 50 °C for 30 s, and 68 °C for 1 min and then 72 °C for 7 min before holding the products at 4 °C.