Table 1 PCR methods used, respective genera and species-specific primers, their target genes, expected PCR product sizes, and amplification conditions.
From: Prevalence and seasonal variation of tick-borne haemoparasites in cattle from north-eastern Uganda
Pathogen | Diagnostic method | Target gene | Primers | PCR product size (Bp) | Reaction conditions | Reference |
|---|---|---|---|---|---|---|
Theileria /Babesia spp. | Conventional PCR | 18S rDNA | F: 5′-GACACA GGAGGTAGTGA CAAG-3′ | 460–540 | 94 °C 5 min, 30 × (94 °C 30 s, 52 °C 45 s, 72 °C 1 min), 72 °C 7 min | |
R: 5′-CTAAGA ATTTCA CCTCTG ACAGT-3′ | ||||||
Anaplasma/Ehrlichia spp. | Conventional PCR | 16S rDNA | F: 5′-GGAATTCAGAGTTGGATCMTGGYTCAG-3′ | 492–498 | 95 °C 5 min, 35 × (94 °C 1 min, 61 °C 1 min, 72 °C 1 min), 72 °C 7 min | |
R: 5′-CGGGATCCCGAGTTTGCGGGACT TYT TCT-3′ | ||||||
B. bigemina | Nested PCR 1 | RAP-1a | F: 5′-GAGTCTGCCAAATCCTTA C-3′ | 879 | 95 °C 5 min, 35 × (94 °C 1 min, 50 °C 1 min, 72 °C 75 s), 72 °C 10 min | |
R: 5′-TCCTCTACAGCTGCTTCG-3′ | ||||||
Nested PCR 2 | F: 5′-AGCTTGCTTTCACAACTCGCC-3′ | 412 | 95 °C 3 min, 35 × (94 °C 1 min, 50 °C 1 min, 72 °C 1 min), 72 °C 7 min | |||
R: 5′-TTGGTGCTTTGACCGACGACAT-3′ | ||||||
B. bovis | Nested PCR 1 | SBP-2 | F: 5′-CTGGAAGTGGATCTCATG CAACC-3′ | 1236 | 95 °C 5 min, 35 × (94 °C 1 min, 55 °C 1 min, 72 °C 90 s), 72 °C 7 min | |
R: 5′-TCACGAGCACTCTACGGCTTTGCAG-3′ | ||||||
Nested PCR 2 | F: 5′-GAATCTAGGCATATAAGGCAT-3′ | 580 | 94 °C 3 min, 35 × (94 °C 1 min, 50 °C 1 min, 72 °C 1 min), 72 °C 7 min | |||
R: 5′-ATCCCCTCCTAAGGTTGGCTAC-3′ | ||||||
T. parva | Nested PCR 1 | p104 | F: 5′-ATTTAAGGAACCTGACGTGACTGC-3′ | 496 | 95 °C 5 min, 35 × (94 °C 1 min, 58 °C 1 min, 72 °C 1 min), 72 °C 9 min | |
R: 5′-TAAGATGCCGACTATTAATGACACC-3′ | ||||||
Nested PCR 2 | F: 5’-GGCCAAGGTCTCCTTCAGAATACG-3’ | 277 | 94 °C 1 min, 35 × (94 °C 1 min, 60 °C 1 min, 72 °C 1 min), 72 °C 9 min | |||
R: 5′-TGGGTGTGTTTCCTCGTCATCTGC-3′ | ||||||
A. marginale | Conventional PCR | MSP-4 | F: 5′-GGGAGCTCCTATGAATTACAGAGAATTGTTTAC-3′ | 851 | 95 °C 5 min, 35 × (94 °C 1 min, 59 °C 1 min, 72 °C 75 s), 72 °C 7 min | |
R: 5′-CCGGATCCTTAGCTGAACAGGAATCTTGC-3′ | ||||||
E. ruminantium | Conventional PCR | pCS20 | F: 5′-CCCTATGATACAGAAGGTAACCTCGC-3′ | 903 | 95 °C 5 min, 35 × (94 °C 1 min, 52 °C 1 min, 72 °C 90 s), 72 °C 10 min | |
R: 5′-GATAAGGAGATAACGTTTGTTTGG-3′ |
