TABLE 2.
PCR programs used for the detection and identification of Babesia spp. in ticks
| PCR | Program | Primer
|
||
|---|---|---|---|---|
| Designation | Sequence (5′-3′) | Approximate nucleotide position | ||
| 18S rRNA gene | 95°C for 1 min, 53°C for 1.50 min, | Babfor | GACTAGGGATTGGAGGTC | 970 |
| and 72°C for 1.50 min for 35 cycles | Babrev | GAATAATTCACCGGATCACTC | 1590 | |
| ITS1 | 95°C for 1 min, 52°C for 2 min, and | ITS1for | CGAGTGATCCGGTGAATTATTC | 1590 (18S rRNA gene) |
| 72°C for 3 min for 40 cycles | ITS1rev | CCTTCATCGTTGTGTGAGCC | Within 5.8S rRNA gene | |
| ITS2 and HSP70 | 95°C for 1 min, 54°C for 2 min, and | ITS2for | GGCTCACACAACGATGAAGG | Within 5.8S rRNA gene |
| 72°C for 3 min for 40 cycles | ITS2rev | CTCGCCGTTACTAAGGGAATC | 76 (28S rRNA gene) | |
| HSP70for | GCTATTGGTATTGACTTGGG | 13 | ||
| HSP70rev | CCTTCATCTTGATAAGGACC | 366 | ||
| Sequencing | 96°C for 10 s, 50°C for 5 s, and 60°C for 4 min for 30 cycles | |||