Table 1.
PCR targets and primers used in this study for detection of Babesia spp., Bartonella spp., Ehrlichia/Anaplasma spp., H. canis, hemotropic Mycoplasma spp. and Rickettsia spp.
| Species | Target gene | Forward primer (5′–3′) | Reverse primer (5′–3′) | Final primer concentration (μM) | Reference |
|---|---|---|---|---|---|
| Babesia spp. | 18S rRNA | GACGATCAGATACCGTCGTAGTCC | CAGAACCCAAAGACTTTGATTTCTCTC | 0.3 | [62] |
| Bartonella spp. | ITS1 | AGATGATGATCCCAAGCCTTCTG | CCTCCGACCTCACGCTTATCA | 0.3 | Modified from [63, 64] |
| Ehrlichia spp.; Anaplasma spp. | 16S rRNA | GCAAGCYTAACACATGCAAGTCG | GGATTATACAGTATTACCCAYCATTTCTARTG | 0.5 | [65, 66] |
| H. canis | 18S rRNA | CTTACCGTGGCAGTGACGGT | ATTGTTATTTCTTGTTACTACCTCTCTCAAAC | 0.3 | [66] |
| Hemotropic Mycoplasma spp. | 16S rRNA | ATGTTGCTTAATTCGATAATACACGAAA | ACRGGATTACTAGTGATTCCAACTTCAA | 0.3/0.5 | [65] |
| Rickettsia spp. | ITS2 | GCTCGATTGRTTTACTTTGCTGTGAG | CATGCTATAACCACCAAGCTAGCAATAC | 0.5/0.3 | [62] |
Abbreviations: rRNA ribosomal ribonucleic acid, ITS internal transcribed spacer