Table 1 Primers and polymerase chain reaction conditions for detection of pathogens in Ixodes, R. sanguineus and D. reticulatus
From: Ticks and associated pathogens collected from dogs and cats in Belgium
Organism detected | Target gene | Primer sequences (5’-3’) | Product t size (bp) | Melting temperature | Reference |
|---|---|---|---|---|---|
Borrelia burgdorferi s.l. | Flagellin | Outer primers: | 236 | • Denaturation 95°C 10 min | / |
Bflag2F: GCT GAA GAR CTT GGA ATG CAR CC | • Hybridisation 35 cycles: 95°C 30s, 59°C 30s, 72°C 30s | ||||
Bflag2R: AGC AGG YGY TGG YTG YTG AGC | |||||
• Extension 72°C 10 min | |||||
Inner primers: | 193 | • Denaturation 95°C 10 min | / | ||
Bflag2nestF: CWC CAG CRT CAC TTT CAG GR | • Hybridisation 35 cycles: 95°C 30s, 56°C 30s, 72°C 30s | ||||
• Extension 72°C 10 min | |||||
Bflag2nestR: GYT GGY TGY TGA GCT CCT TC | |||||
Anaplasma phagocytophilum | Major surface protein 2 (msp2) | Apmsp2f: ATG GAA GGT AGT GTT GGT TAT GGT ATT | 77 | • Denaturation 95°C 10 min | Courtney et al., 2004 |
Apmsp2r TTG GTC TTG AAG CGC TCG TA | |||||
• Hybridisation 40 cycles: 95°C 15 s, 60°C 1 min | |||||
• Extension 60°C 1 min | |||||
Rickettsia spp. | Citrate synthase (gltA) | Rsfg877: GGG GGC CTG CTC ACG GCG G | 380 | • Denaturation 95°C 10 min | Reis et al., 2011 |
Rsfg1258: ATT GCA AAA AGT ACA GTG AAC A | • Hybridisation 5 cycles: 95°C 60s, 58°C 60s, 72°C 60s | ||||
35 cycles: 95°C 60s, 51°C 60s, 72°C 60s | |||||
• Extension 72°C 10 min | |||||
Babesia spp. | 18S rRNA | BJ1:GTC TTG TAA TTG GAA TGA TGG | 411-452 | • Denaturation 94°C 10 min | Casati et al, 2006 |
|  |  | BN2:TAG TTT ATG GTT AGG ACT ACG | • Hybridisation: 35 cycles 94°C 1 min, 55°C 1 min, 72° 2 min | ||
|  |  |  |  | • Extension 72°C 5 min |