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Research Article

Table 1

Primers used in the PCR assays conducted in this study with their respective references.

Organism Targeted gene (product size) Name and primer sequences (5′ – 3′) PCR conditions Ref.
Tick Mitochondrial 16S rDNA (320 bp) TQ 16S+1F: Denaturation [2]
 5′-CTGCTCAATGATTTTTTAAATTGCTGTGG-3′  94 °C 5 min
TQ 16S−2R: Hybridization
 5′-ACGCTGTTATCCCTAGAG-3′  10 cycles: 92 °C 1 min; 48 °C 1 min; 72 °C 1 min 30 s
 32 cycles: 92 °C 1 min; 54 °C 1 min; 72 °C 1 min 30 s
Extension
 72 °C 5 min
Anaplasmataceae 16S rRNA (345 bp) EHR 16SD: Denaturation [36]
 5′-GGTACCYACAGAAGAAGTCC-3′  94 °C 5 min
EHR 16SR: Hybridization
 5′-TAGCACTCATCGTTTACAGC-3′  34 cycles: 94 °C 40 s; 50 °C 40 s; 72 °C 1 min
Extension
 72 °C 10 min
Anaplasma phagocytophilum 16S rDNA (932 bp) ge3a: Denaturation [28]
 5′-CACATGCAAGTCGAACGGATTATTC-3′  95 °C 5 min
ge10r: Hybridization
 5′-TTCCGTTAAGAAGGATCTAATCTCC-3′  40 cycles: 94 °C 30 s; 55 °C 30 s; 72 °C 1 min
Extension
 72 °C 5 min
16S rDNA (546 bp) ge9f: Denaturation
 5′-AACGGATTATTCTTTATAGCTTGCT-3′  95 °C 5 min
ge2r: Hybridization
 5′-GGCAGTATTAAAAGCAGCTCCAGG-3′  30 cycles: 94 °C 30 s; 55 °C 30 s; 72 °C 1 min
Extension
 72 °C 5 min
Borrelia burgdorferi s.l. 16S rRNA (357 bp) 16S LDF: Denaturation [27]
 5′-ATGCACACTTGGTGTTAACTA-3′  95 °C 5 min
16S LDR: Hybridization
 5′-GACTTATCACCGGCAGTCTTA-3′  35 cycles: 95 °C 1 min; 53 °C 1 min; 72 °C 1 min
Extension
 72 °C 10 min
Babesia/Theileria spp. 18S rRNA (359 bp) BAB GF2: Denaturation [4]
 5′-GYYTTGTAATTGGAATGATGG-3′  94 °C 5 min
BAB GR2: Hybridization
 5′-CCAAAGACTTTGATTTCTCTC-3′  35 cycles: 94 °C 1 min; 60 °C 1 min; 72 °C 1 min
Extension
 72 °C 10 min
Rickettsia spp. gltA (citrate synthase) (381 bp) RpCS.877p: Denaturation [39]
 5′-GGGGGCCTGCTCACGGCGG-3′  95 °C 5 min
RpCS.1258n: Hybridization
 5′-ATTGCAAAAAGTACAGTGAACA-3′  6 cycles: 94 °C 1 min; 60 °C 1 min; 72 °C 1 min
 30 cycles: 94 °C 1 min; 53 °C 1 min; 72 °C 1 min
Extension
 72 °C 10 min
SFG Rickettsia ompA (532 bp) Rr190.70p: Denaturation [39]
 5′-ATGGCGAATATTTCTCCAAAA-3′  95 °C 5 min
Rr190.602n: Hybridization
 5′-AGTGCAGCATTCGCTCCCCCT-3′  6 cycles : 94 °C 1 min; 60 °C 1 min; 72 °C 1 min
 30 cycles : 94 °C 1 min; 53 °C 1 min; 72 °C 1 min
Extension
 72 °C 10 min

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