Table 1.

Primers and PCR conditions used for the detection of the five pathogen groups^a

Pathogen	Primer	Orientation	Target gene	Sequence (5′–3′)	Reference	[Primer] (μM)	[MgCl2] (mM)	Annealing temp (°C)	Elongation time (s)
Anaplasmataceae	EHR1	Forward	16S rRNA	GAACGAACGCTGGCGGCAAGC	29	0.4	2	63	45
	newEHR2b	Reverse	16S rRNA	CACGCTTTCGCACCTCAGTGTC	29	0.4
	EHR3	Forward	16S rRNA	TGCRTAGGAATCTRCCTAGTAG	29	0.8	2	59	45
	newEHR2b	Reverse	16S rRNA	CACGCTTTCGCACCTCAGTGTC	29	0.8
Rickettsiaceae	Rr17k.1p	Forward	17 kDa	TTTACAAAATTCTAAAAACCAT	10	0.8	2	55	45
	Rr17k.539n	Reverse	17 kDa	TCAATTCACAACTTGCCATT	10	0.8
	Rr17k.90p	Forward	17 kDa	GCTCTTGCAACTTCTATGTT	10	0.8	2	54	45
	Rr17k.539n	Reverse	17 kDa	TCAATTCACAACTTGCCATT	10	0.8
Piroplasmidae	BJ1	Forward	18S rRNA	GTCTTGTAATTGGAATGATGG	5	0.8	3	61	60
	BN2	Reverse	18S rRNA	TAGTTTATGGTTAGGACTACG	5	0.8
Coxiella	Q5	Forward	htpB	GCGGGTGATGGTACCACAACA	35	0.4	1.5	58	30
	Q3	Reverse	htpB	GGCAATCACCAATAAGGGCCG	35	0.4
	Q6	Forward	htpB	TTGCTGGAATGAACCCCA	35	0.8	2	56	30
	Q4	Reverse	htpB	TCAAGCTCCGCACTCATG	35	0.8
Borrelia species	newLDfb	Forward	16S rRNA	GTAAACGATGCACACTTGGTG	14	0.4	2	61	30
	newLDrb	Reverse	16S rRNA	TCCGRCTTATCACCGGCAGTCT	14	0.4
	Outer1	Forward	flaB	AARGAATTGGCAGTTCAATC	6	0.8	2	59	30
	Outer2	Reverse	flaB	GCATTTTCWATTTTAGCAAGTGATG	6	0.8
	Inner1	Forward	flaB	ACATATTCAGATGCAGACAGAGGTTCTA	6	0.8	2	59	30
	Inner2	Reverse	flaB	GAAGGTGCTGTAGCAGGTGCTGGCTGT	6	0.8

^aPCR protocol: 94°C for 3 min; 40 cycles of 94°C for 30 s; specific annealing at 30 s and 72°C for specific elongation; subsequent incubation at 72°C for 10 min.

^bThe primer sequence was modified.