. 2022 May 16;16(5):e0010437. doi: 10.1371/journal.pntd.0010437

Table 1. The primers, probes, and thermocycling conditions^a that were used to detect Rickettsia africae, Rickettsia, and tick DNA in a novel multiplex qPCR assay for this study.

Target organism	Target gene	Primer name	Primer orientation	5’-3’ sequence	Reference	Concentration (nM)
Tick	16S	Tick-F2	Forward	CTCTAGGGATAACAGCGTWATAWT	This study	600
		Tick-R	Reverse	GTCTGAACTCAGATCAAGTAGG		600
		Tick-P1	Probe	6-FAM/TGCGACCTC/ZEN/GATGTTGGATTAGGA/3’IowaBlack		250
Amblyomma hebraeum	CO1	Aheb_F	Forward	CATCATAATTGGCGGGTTTG	This study	400
		Aheb_R	Reverse	AGTAAACAAAGAGATGGTGGTA		400
		Aheb_P	Probe	Cy5/TGACTAGTT/TAO/CCAATTATGCTAGGTGCCC/3’IowaBlack		250
Rickettsia africae	ompB	Raf1797F	Forward	TTGGAGCTAATAATAAAACTCTTGGAC	[25]	100
		Raf1915R	Reverse	GAATTGTACTGCACCGTTATTTCC		100
		Raf1879P	Probe	HEX/CGCGATGTTAATAGCAACATCACC GCCACTATCGCG/Black Hole Quencher		250

^aThermocycling conditions were as follows: initial denaturation of 95°C for 15 minutes followed by 40 cycles of 95°C for 45 seconds and 60°C for 90 seconds.

Table 1. The primers, probes, and thermocycling conditionsa that were used to detect Rickettsia africae, Rickettsia, and tick DNA in a novel multiplex qPCR assay for this study.

Table 1. The primers, probes, and thermocycling conditions^a that were used to detect Rickettsia africae, Rickettsia, and tick DNA in a novel multiplex qPCR assay for this study.