Table 1.
Pathogen | Primer | Orientation | Target gene | Sequence (5′–3′) | Reference | [Primer] (μM) | [MgCl2] (mM) | Annealing temp (°C) | Elongation time (s) |
---|---|---|---|---|---|---|---|---|---|
Anaplasmataceae | EHR1 | Forward | 16S rRNA | GAACGAACGCTGGCGGCAAGC | 29 | 0.4 | 2 | 63 | 45 |
newEHR2b | Reverse | 16S rRNA | CACGCTTTCGCACCTCAGTGTC | 29 | 0.4 | ||||
EHR3 | Forward | 16S rRNA | TGCRTAGGAATCTRCCTAGTAG | 29 | 0.8 | 2 | 59 | 45 | |
newEHR2b | Reverse | 16S rRNA | CACGCTTTCGCACCTCAGTGTC | 29 | 0.8 | ||||
Rickettsiaceae | Rr17k.1p | Forward | 17 kDa | TTTACAAAATTCTAAAAACCAT | 10 | 0.8 | 2 | 55 | 45 |
Rr17k.539n | Reverse | 17 kDa | TCAATTCACAACTTGCCATT | 10 | 0.8 | ||||
Rr17k.90p | Forward | 17 kDa | GCTCTTGCAACTTCTATGTT | 10 | 0.8 | 2 | 54 | 45 | |
Rr17k.539n | Reverse | 17 kDa | TCAATTCACAACTTGCCATT | 10 | 0.8 | ||||
Piroplasmidae | BJ1 | Forward | 18S rRNA | GTCTTGTAATTGGAATGATGG | 5 | 0.8 | 3 | 61 | 60 |
BN2 | Reverse | 18S rRNA | TAGTTTATGGTTAGGACTACG | 5 | 0.8 | ||||
Coxiella | Q5 | Forward | htpB | GCGGGTGATGGTACCACAACA | 35 | 0.4 | 1.5 | 58 | 30 |
Q3 | Reverse | htpB | GGCAATCACCAATAAGGGCCG | 35 | 0.4 | ||||
Q6 | Forward | htpB | TTGCTGGAATGAACCCCA | 35 | 0.8 | 2 | 56 | 30 | |
Q4 | Reverse | htpB | TCAAGCTCCGCACTCATG | 35 | 0.8 | ||||
Borrelia species | newLDfb | Forward | 16S rRNA | GTAAACGATGCACACTTGGTG | 14 | 0.4 | 2 | 61 | 30 |
newLDrb | Reverse | 16S rRNA | TCCGRCTTATCACCGGCAGTCT | 14 | 0.4 | ||||
Outer1 | Forward | flaB | AARGAATTGGCAGTTCAATC | 6 | 0.8 | 2 | 59 | 30 | |
Outer2 | Reverse | flaB | GCATTTTCWATTTTAGCAAGTGATG | 6 | 0.8 | ||||
Inner1 | Forward | flaB | ACATATTCAGATGCAGACAGAGGTTCTA | 6 | 0.8 | 2 | 59 | 30 | |
Inner2 | Reverse | flaB | GAAGGTGCTGTAGCAGGTGCTGGCTGT | 6 | 0.8 |
PCR protocol: 94°C for 3 min; 40 cycles of 94°C for 30 s; specific annealing at 30 s and 72°C for specific elongation; subsequent incubation at 72°C for 10 min.
The primer sequence was modified.