TABLE 5.
PCR primers used for tick identification, detection of Borrelia spp., and identification of blood meal sources
| Primer | Target gene | Target species | 5′–3′ primer sequence | Amplicon length (bp) | Reference no. or source |
|---|---|---|---|---|---|
| Primers for tick identification | |||||
| 16S+1 | 16S rRNA | Ticks | CTGCTCAATGATTTTTTAAATTGCTGTGG | 480 | 47 |
| 16S-1 | 16S rRNA | Ticks | CCGGTCTGAACTCAGATCAAGT | 480 | 47 |
| Primers for Borrelia sp. identification | |||||
| Bfpbu | flaB | Borrelia | GCTGAAGAGCTTGGAATGCAACC | 346 | 29 |
| Bfpcr | flaB | Borrelia | TGATCAGTTATCATTCTAATAGCA | 346 | 29 |
| BOR1 | flaB | Borrelia | TAATACGTCAGCCATAAATGC | 750 | 30 |
| BOR2 | flaB | Borrelia | GCTCTTTGATCAGTTATCATTC | 750 | 30 |
| GlpQ-510f | glpQ | Borrelia | AAAACCCTTTTGGCATAAACAACA | 280 | 31 |
| GlpQ-770r | glpQ | Borrelia | CCAGGGTCCAATTCCGTCAG | 280 | 31 |
| Primers for identification of blood meal | |||||
| 12S-16SF | 12S and 16S rRNA | All vertebrates | ACACCGCCCGTCACCCTCC | 500 | 27 |
| 12S-16SR | 12S and 16S rRNA | All vertebrates | AACCAGCTATCACCAGGCTCG | 500 | 27 |
| 16S24F | 16S rRNA, inner primer for nested PCR | All vertebrates | AAGTCGTAACATGGTAAGCA | 366 | This study |
| 16S366R | 16S rRNA, inner primer for nested PCR | All vertebrates | GGTAGCTCGTCTGGTTTCGG | 366 | This study |
| Por100f | 16S rRNA | Porcupine | CAGAAGACTTCAGGTAATACTGGC | 311 | This study |
| Por391r | 16S rRNA | Porcupine | GGGTTGGCTCAATTTAGCTGT | 311 | This study |
| FOX131f | 16S rRNA | Fox | ACGAAAGCTAGCCCAATCGAC | 303 | This study |
| FOX410r | 16S rRNA | Fox | CACTATTTTGCCACATAGACGAGT | 303 | This study |
| BADG45f | 16S rRNA | Badger | ACATCATGACCACTTTGAACCA | 302 | This study |
| BADG345r | 16S rRNA | Badger | TGAGTTTATCCCTGTGGATTGT | 302 | This study |
| Jack134f | 16S rRNA | Jackal | GCTAGCCCAACTAACCCCAA | 287 | This study |
| Jack401r | 16S rRNA | Jackal | TTTTGCTACATAGATGAGTTGATCC | 287 | This study |
| Hyrax102f | 16S rRNA | Hyrax | AACCCCATTGACCACTTTGAACT | 305 | This study |
| Hyrax407r | 16S rRNA | Hyrax | GGGTTGGTTCTGTAAACTGCTC | 305 | This study |
| Cat95f | 16S rRNA | Cat | TCATATTAAACTGACCATCTTGAGC | 318 | This study |
| Cat412r | 16S rRNA | Cat | CAACATAGACGAGTTCATCCTGTAA | 318 | This study |
| goat 162F | 16S rRNA | Goat | TTACCAAAACAGTCTAAAACAA | 238 | This study |
| Goat380r | 16S rRNA | Goat | AACTGCCCATGAGTAGCTCG | 238 | This study |
| Unrev1025 | Cytochrome b | Mammal | GGTTGTCCTCCAATTCATGTTA | 334/680c | 26 |
| Human741Fa | Cytochrome b | Human | GGCTTACTTCTCTTCATTCTCTCCT | 334 | 26 |
| Dog368Fa | Cytochrome b | Dog | GGAATTGTACTATTATTCGCAACCAT | 680 | 26 |
| 12S-13R | 12S rRNA | All vertebrates | AGGAGGGTGACGGGCGGT | 326/369d | 24 |
| RodentFb | 12S rRNA | Muridae, Cricetidae | GGCGGTACTTTATATCCAT | 326 | 24 |
| BirdFb | 12S rRNA | Birds | TACGAGCACAAACGCTTAA | 369 | 24 |
a
Forward primers used with UNREV1025.
b
Forward primers used with 12S-13R.
c
Reverse primer UNREV amplifies a 334-bp amplicon with the forward primer Human741F when amplifying a human blood meal and a 680-bp amplicon with the forward primer Dog368F when amplifying a dog blood meal.
d
Reverse primer 12S-13R amplifies a 326-bp amplicon with the forward primer RodentF when amplifying a rodent blood meal and a 369-bp amplicon with the forward primer BirdF when amplifying a bird blood meal.