From: Eurasian golden jackal as host of canine vector-borne protists
Parasite | Gene | Fragment length (bp) | Primer | Primer sequence (5’-3’) | PCR conditions, master mix used | Source |
---|---|---|---|---|---|---|
Hepatozoon spp. | 18S | ~670 | Hep F | ATACATGAGCAAAATCTCAAC | 94 °C for 3 min, 35× (94 °C for 1 min, 61 °C for 1 min, 72 °C for 2 min), 72 °C for 7 min | [17] |
Hep R | CTTATTATTCCATGCTGCAG | PPP Master Mix (TopBio s.r.o., Czech Republic) | ||||
18S | ~1765 | HAM 1 F | GCCAGTAGTCATATGCTTGTC | 95 °C for 5 min, 35× (95 °C for 20 s, 56 °C for 1 min, 72 °C for 1 min), 72 °C for 5 min | [18] | |
HPF 2R | GACTTCTCCTTCGTCTAAG | PCRBIO Taq DNA Polymerase (PCR Biosystems Ltd, UK) | ||||
Piroplasmida | 18S | ~685 | BTH 1 F | CCTGAGAAACGGCTACCA CATCT | 95 °C for 10 min, 40× (95 °C for 30 s, 60 °C for 1 min, 72 °C for 1 min), 72 °C for 10 min | [19] |
BTH 1R | TTGCGACCATACTCCCCCCA | |||||
~560 | GF2 | GTCTTGTAATTGGAATGA TGG | 95 °C for 10 min, 40× (95 °C for 30 s, 62 °C for 1 min, 72 °C for 1 min), 72 °C for 10 min | |||
GR2 | CCAAAGACTTTGATTTCTCTC | PPP Master Mix (TopBio s.r.o., Czech Republic) | ||||
18S | ~1730 | BT1 F | GGTTGATCCTGCCAGTAGT | 94 °C for 30 s, 20× (94 °C for 30 s, 65-55 °C (-0.5 °C/cycle) for 30 s, 68 °C for 1 min), 20× (94 °C for 30 s, 55 °C for 30 s, 68 °C for 1 min), 68 °C for 5 min | [20] | |
BT outer R | GGAAACCTTGTTACGACTTCTC | |||||
~1670 | Piro0F2 | GCCAGTAGTCATATGCTTGTCTTA | 94 °C for 30 s, 20× (94 °C for 30 s, 65-55 °C (-0.5 °C/cycle) for 30 s, 68 °C for 1 min), 20× (94 °C for 30 s, 55 °C for 30 s, 68 °C for 1 min), 68 °C for 5 min | [21] | ||
BT Inner R | TTC TCC TTC CTT TAA GTG ATA AG | OneTaq 2x Master Mix with standard buffer (NEB Inc., USA) | [20] | |||
Babesia spp. | cox1 | ~1250 | Bab_For1 | ATWGGATTYTATATGAGTAT | 95 °C for 1 min, 35× (95 °C for 15 s, 45 °C for 30 s, 72 °C for 30 s), 72 °C for 10 min | [22] |
Bab_Rev1 | ATAATCWGGWATYCTCCTTGG | modified | ||||
~975 | Bab_For2 | TCTCTWCATGGWTTAATTATGATAT | 95 °C for 1 min, 35× (95 °C for 15 s, 45 °C for 30 s, 72 °C for 30 s), 72 °C for 10 min | |||
Bab_Rev2 | TAGCTCCAATTGAHARWACAAAGTG | PCRBIO Taq DNA Polymerase (PCR Biosystems Ltd, UK) | ||||
Theileria spp. | cox1 | ~1020 | Cox1F133 | GGAGAGCTAGGTAGTAGTGGAGATAGG | 95 °C for 1 min, 35× (95 °C for 15 s, 63 °C for 15 s, 72 °C for 30 s), 72 °C for 10 min | [21] |
Cox1R1130 | GTGGAAGTGAGCTACCACATACGCTG | |||||
~985 | Cox_cladeI_Fw2 | GTAGTGGAGATAGGTTCATAGC | 95 °C for 1 min, 35× (95 °C for 15 s, 55 °C for 15 s, 72 °C for 30 s), 72 °C for 10 min | This study | ||
Cox_cladeI_Rev2 | TGTATCGTGTAGTGACACGTC | PCRBIO Taq DNA Polymerase (PCR Biosystems Ltd, UK) | ||||
Leishmania spp. | ITS1 | ~280 | ITS-219 F | AGCTGGATCATTTTCCGATG | qPCR: 95 °C for 5 min, 45× (95 °C for 5 s, 57 °C for 15 s, 72 °C for 15 s), melting from 60 to 95 °C at 1 °C/s | [26] |
ITS-219R | ATCGCGACACGTTATGTGAG | iQSYBER Green Supermix (BioRad Inc., USA) | ||||
ITS1-5.8S | ~300 | LITSR | CTGGATCATTTTCCGATG | 95 °C for 3 min, 40× (95 °C for 20 s, 53 °C for 35 s, 72 °C for 60 s), 72 °C for 5 min | [27] | |
L5.8S | ACACTCAGGTCTGTAAAC | |||||
EmeraldAmp (TaKaRa Clontech, Japan) |