<i>Ehrlichia muris</i> in <i>Ixodes cookei</i> Ticks, Northeastern United States, 2016–2017



Table 1 .

 1 Ehrlichia muris in Ixodes cookei Ticks, Northeastern United States, 2016-2017 Primers and probes used in this study 


	Technical Appendix			
	Technical Appendix Target				
	gene	Application	Type	Sequences (5′3′)	Tm (C)	Reference
	16S	Tick species	Forward	TGCTGTAGTATTTTGACTATACAAAGG	55	This paper
		PCR and	Reverse	ATCCTAATCCAACATCGAGGTC		
		confirmation				
	ITS	Ixodes	Forward	TGCGTTTTCTTTGAGCAAATGCACGAG	60	This paper
		scapularis	Reverse	GTACGGGATTTTCCACAAACGGTATCCA		
		identification	Probe	FAM-TGCGCTTAACCAGTCCTCCTCCTCCTACGA-BHQ		
	ITS	Ixodes pacificus	Forward	CTCGGAGCAAGTACGGAGGTAG	60	This paper
		identification	Reverse	TTTCCACAAAACGGTCGCCATC		
			Probe	Cy5-CTGAGCCAAGTCCTCTTCCTACCCGGTTTG-BHQ		
	ITS	Amblyomma	Forward	CGACCGCCGCAGGAAGG	60	This paper
		americanum	Reverse	CGTTTCTCGCAGCAGTTCGG		
		identification	Probe	FAM-CCCGCTGGCCCGCGTACGTGT-BHQ		
	ITS	Dermacentor	Forward	CTGAAGATTCTTTGCGAGGAGCGG	60	(1)
		variabilis	Reverse	GCGTCAGCTCGGCCAAC		
		identification	Probe	FAM-AGAAGGGCGTGCCCCGAAAGCGG-BHQ		
	gltA	E. muris PCR	Forward	TACAGATTTCTCAAGAATATACA (outer)	50	(2)
		and confirmation		TGGCATGTTTTTCTGCCTTA (inner)		
			Reverse	AATGCAATGTTTTCTAATTCTAC (outer)		
				TGACCAAAACCCATTAATCTTG (inner)		
	groEL	E. muris PCR	Forward	GGATCCATTGGCTCTTGCTA (outer)	50	(2)
		and confirmation		AAGGGATTCAAAGAATTGGATG (inner)		
			Reverse	CCACCAACCTTTAAGACAGCA (outer)		
				CCACCAACCTTTAAGACAGCA (inner)		
	P13	EMLA detection	Forward	TACCTAATTCTTCTCAAGAGATTCAGTTG	60	This paper
			Reverse	ATGATGATACTGCGAACAACTATAAGAG		
			Probe	Cy5-		
				ATATTGATAAAAGAGTCAGTGTTGATCCGTATGAGTTA		
				GGGTT-BHQ		
	glpQ	Borrelia	Forward	GACATAGTTCTAACAAAGGACAATATTCC	60	(3)
		miyamotoi	Reverse	TCCGTTTTCTCTAGCTCGATTGG		
		detection	Probe	HEX-TGCACGACCCAGAAATTGACACAACCACAA-BHQ		
	ospA	Borrelia	Forward	ATAGGTCTAATATTAGCCTTAATAGCAT	60	This paper
		burgdorferi	Reverse	AGATCGTACTTGCCGTCTT		
		Sensu Lato	Probe	FAM-aagc+Aaa+Atgtt+Agc+Agccttga-BHQ (LNA)		
		detection				
	Tubulin	Babesia	Forward	GATTTGGAACCTGGCACCATG	60	(4)
		detection	Reverse	AATGACCCTTAGCCCAATTATTTCC		
			Probe	FAM-ATCTGGCCCATACGGTGAATTGTTTCGC-BHQ		
	MSP-2	Anaplasma	Forward	ATGGAAGGTAGTGTTGGTTATGGTATT	60	(4)
		detection	Reverse	TTGGTCTTGAAGCGCTCGTA		
			Probe	HEX-TGGTGCCAGGGTTGAGCTTGAGATTG-BHQ		
	*We use species-specific taqman qPCR identified Amblyomma americanum (243), Dermacentor variabilis (271), Ixodes pacificus (215), and Ixodes
	scapularis (7800). A fragment of 16S mtDNA was used to identify EMLA-positive Ixodes scapularis ticks (2), Ixodes angustus (14), Ixodes cookei
	(22), Ixodes ricinus (5), Ixodes spinipalpis (19) and Dermacentor occidentalis (3). The rest 114 ticks were identified by morphological characters or
	marked as unidentifiable because of poor sample conditions.		

Technical AppendixTable 3 .

 3 EMLA positive ticks in this study 


	Tick ID#	Tick species	Tick stage	Source	Location
	20170522-362	Ixodes scapularis	Adult	Human	Laporte, MN 56461
	20170605-176	Ixodes scapularis	Adult	Human	Eleva, WI 54738
	20170731-52	Ixodes cookei	Adult	Human	Holden, ME 04429
	20170807-147	Ixodes cookei	Adult	Human	Salamanca, NY 14779
	20170918-13	Ixodes cookei	Adult	Human	Littleton, ME 04730
Guang Xu, Patrick Pearson, Stephen M Rich
<i>Ehrlichia muris</i> in <i>Ixodes cookei</i> Ticks, Northeastern United States, 2016–2017