Table 1.

Description of primers, hydrolysis probes and thermal sequences used in qPCR assays for Ehrlichia spp. and Anaplasma spp.

Agents (target-genes)	Primers	Hydrolysis probe (TaqMan)	Thermal sequences	References
Ehrlichia spp. (groEL gene)a	5′-GCGAGCATAATTACTCAGAG-3’ 5′-CAGTATGGAGCATGTAGTAG-3’	TET-5′-CATTGGCTCTTGCTATTGCTAAT- 3′[BHQ2a-Q]3′	− 95 °C for 3 min; 40 cycles: 95 °C for 10 min and 52.8 °C for 30 s	18
Anaplasma spp. (groEL gene)a	5′-TTATCGTTACATTGAGAAGC-3′ 5′GATATAAAGTTATTAAAAGTATAAAGC-3′	Cy-5-5′-CCACCTTATCATTACACTGAGACG-3′[BHQ2a-Q]3′	− 95 °C for 3 min; 40 cycles: 95 °C for 10 min and 52.8 °C for 30 s	18
E. canis (dsb gene)	5′-TTGCAAAATGATGTCTGAAGATATGAAACA-3′ 5′-GCTGCTCCACCAATAAATGTATCYCCTA-3′	5′ FAM AGCTAGTGCTGCTTGGGCAACTTTGAGTGAA-[ BHQ-1–3′]	95 °C for 5 min, 40 cycles: 95 °C for 15 s and 60 °C for 1 min	34
E. chaffeensis (vlpt gene)	5′-CTAATTCTGATTTACACGAGTCTTC-3′ 5′-GCATCATCTTCGAATTGAACTTC-3′	5′[TAMRA] (TTGAGTGTCC[BHQ2a-Q]3′	95 °C for 3 min, 40 cycles: 95 °C for 10 min e 55 °C for 30 s	35
A.phagocytophilum (msp-2 gene)	5′-AGTTTGACTGGAACACACCTGATC-3 5′-CTCGTAACCAATCTCAAGCTCAAC-3′	5′[FAM] (939p-TTAAGGACAACATGCTTGTAGCTATGGAAG-GCA-[TAMRA])	50 °C for 2 min, 95 °C for 10 min, 40 cycles: 95 °C for 15 s and 60 °C for 1 min	31
A. marginale (msp1-β gene)	5′-TTGGCAAGGCAGCAGCTT-3′ 5′-TTCCGCGAGCATGTTGCAT-3′	5′[FAM] (TCGGTCTAACATCTCCAGGCTTTCAT-3′-BHQ1	95 °C for 10 min, 40 cycles: 95 °C for 15 s and 60 °C for 1 min	32

^aqPCR multiplex.