Table 2 Apicomplexa and Kinetoplastida primers used for cross-validation of NGS results.
Apicomplexa Taxon Targeted | Primer Pair | Gene Targeted | Product Size | Reference |
|---|---|---|---|---|
Canine piroplasm nested PCR | BTF1 & BTR1 BTF2 & BTR2 | 18S rRNA gene | 930 bp 800 bp | |
Hepatozoon canis | HepF & HepR | 18S rRNA gene | 666 bp | |
Coccidia | COC-1 & COC-2 | 18S rRNA gene | 280–350 bp | |
Toxoplasma gondii | Unnamed (see reference) | T. gondii 529 bp repeat element | 529 bp | |
Tissue coccidia | COX10F & COX500R | Cytochrome c oxidase I (COX1) | 470–510 bp | |
Coccidia specific nested PCR | Sarc-int-2F* (5′-AGCTCGTAGTTGGATATCTGCTG-3′) & Sarc-int-2R* (5′-CCTATCTTGTTATTCCATGCTGCA-3′) | Cytochrome c oxidase I (COX1) Uses PCR product from COX10F & COX500R primers | 150 bp | This study* |
Kinetoplastida Taxon Targeted | Primer Pair | Gene Targeted | Product Size | Reference |
Kinetoplastida specific | Kin24SF & Kin24SR | 24S alpha-subunit rRNA | 440–520 bp | |
Trypanosoma evansi | RoTat1.2 F & RoTat1.2 R | T. evansi 1.2 Variable Surface Glycoprotein (VSG) | 205 bp | |
Kinetoplastida specific | KinSSUF1 & KinSSUseqR2 | 18S rRNA | 650 bp | |
Kinetoplastida specific nested PCR | PCaud1F* (5′-CTACCACTTCTACGGAGGGC-3′) & PCaud1R* (5′-GCACCAGACTTGTCCTCCAA-3′) | 18S rRNA. Uses PCR product from KinSSUF1 & KinSSUseqR2 primers | 130 bp | This study* |
