The prevalence of Borrelia miyamotoi infection, and co-infections with other Borrelia spp. in Ixodes scapularis ticks collected in Canada

Table 1 Primer and probe sequences for the detection of A. phagocytophilum and Borrelia species

		Primer/Probe	5′-3′ nucleotide sequence
Duplex screening assay
		Bb23Sf	CGAGTCTTAAAAGGGCGATTTAGT
B. burgdorferi	23S rRNA	Bb23Sr	GCTTCAGCCTGGCCATAAATAG
		Bb23S-P	FAM-AGATGTGGTAGACCCGAAGCCGAGTG-TAMRA
		ApMSP2f	ATGGAAGGTAGTGTTGGTTATGGTATT
A. phagocytophilum	msp2	ApMSP2r	TTGGTCTTGAAGCGCTCGTA
		ApMSP2-P	VIC-TGGTGCCAGGGTTGAGCTTGAGATTG-TAMRA
*B. burgdorferi* confirmatory assay(s)*
		ospAF	CTGGGGAAGTTTCAGTTGAAC
B. burgdorferi	ospA	ospAR	TTGGTGCCATTTGAGTCGTA
		ospA-P	FAM-CTGCAGCTTGGAATTCAGGCACTT-BBQ
		BbIGSf	AAGAAGGACAAGTATTGTAGCGAG
B. burgdorferi	IGS*	BbIGSr	GCAATCTTTGCCTTCCTCC
		BbIGS-P	FAM-TGCCAGTATTTAGTGGTAGGGATTCGG-BBQ
*B. miyamotoi* assays
		BmiyaIGSf	CGTCTTGTTGCTTTTAAAGTGT
B. miyamotoi	IGS	BmiyaIGSr	CATGATCAGGTCCTTGATAATATG
		BmiyaIGS-P	FAM-TGGATTCCAAATTTGATTACATGCAA-BBQ
		MGlpQF	GATAATATTCCTGTTATAATGC
B. miyamotoi	glpq	MGlpQR	CACTGAGATTTAGTGATTTAAGTTC
		MYS-P	FAM-CCCAGAAATTGACAACCACAAATGT-BHQ2
*Borrelia* spp. Nested PCR
16S-23S IGS	1st stage	rrs	GTATGTTTAGTGAGGGGGGTG
		rrl	GGATCATAGCTCAGGTGGTTAG
	2nd stage	Fn	AGGGGGGTGAAGTCGTAACAAG
		Rn	GTCTGATAAACCTGAGGTCGGA
Flagellin	1st stage	FO1	AAGTAGAAAAAGTCTTAGTAAGAATGAAGGA
		FO2	AATTGCATACTCAGTACTATTCTTTATAGAT
	2nd stage	FI1	CACATATTCAGATGCAGACAGAGGTTCTA
		FI2	GAAGGTGCTGTAGCAGGTGCTGGCTGT

*B. burgdorferi IGS real-time PCR is performed solely for validation of the specificity of B. miyamotoi real-time PCR assays and is not routinely used in the testing protocol.

ISSN: 1756-3305