Table 1.
Primers and probes used for Francisella sp. and Acanthamoeba sp. detection.
| qPCR (Target) | Primers/Probes | 5′ to 3′ DNA Sequence | Amplicon Size | Species Detected | Reference |
|---|---|---|---|---|---|
| ISFtu2-qPCR (ISFtu2 insertion sequence element) |
Forward | ttggtagatcagttggtgggataac | 97 bp | Francisella sp. | [22] |
| Reverse | tgagttttaccttctgacaacaatatttc | ||||
| Probe | FAM-aaatccatgctatgactgatgctttaggtaatcca-BHQ1 | ||||
| Tul4-qPCR (tul4 gene) |
Forward | attacaatggcaggctccaga | 91 bp | F. tularensis and F. novicida | [22] |
| Reverse | tgcccaagttttatcgttcttct | ||||
| Probe | FAM-ttctaagtgccatgatacaagcttcccaattactaag-BHQ1 | ||||
| Type B-qPCR (junction between ISFtu2 and a specific flanking 3′ region) |
Forward | cttgtacttttatttggctactgagaaact | 144 bp | F. tularensis subsp. holarctica | [23] |
| Reverse | cttgcttggtttgtaaatatagtggaa | ||||
| Probe | FAM-acctagttcaacc*t*caagacttttagtaatgggaatgtca-BHQ1 Internal quencher | ||||
| Acanth-qPCR (Acanthamoeba sp. 18 S rDNA) |
Forward | cccagatcgtttaccgtgaa | 180 bp | Acanthamoeba sp. | [21] |
| Reverse | taaatattaatgcccccaactatcc | ||||
| Probe | FAM-ctgccaccgaatacattagcatgg-BHQ1 |