Skip to main content

Advertisement

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