Table 3.
Gene targets, primers and probes used in duplex PCR for the LightCycler 2.0 * platform to detect B. anthracis and Bcbva.
| Target | Oligo | Primer/Probe Sequence (5′–3′) | Final Concentration (nM) |
|
|---|---|---|---|---|
| Duplex 1 | Ba-1 | F-primer | GTACATCTTCTAGCTGTTGCAA | 600 |
| R-primer | ACGTAGGAAGACCGTTGATTA | 600 | ||
| Probe | VIC-CGTTGTTGTGTATTTG-MGB | 250 | ||
| GI4 | F-primer | GGAGATATTAACAAGAGATGGATTGGA | 700 | |
| R-primer | CAGTAGGCTTGTCTGCTCTAATAAAATT | 600 | ||
| Probe | FAM-ACATGCCAGCGTTTTTTGCCTCTACACA-BHQ1 | 600 | ||
| Duplex 2 | lef | F-primer | CACTATCAACACTGGAGCGATTCT | 400 |
| R-primer | AATTATGTCATCTTTCTTTGGCTCAA | 400 | ||
| Probe | VIC-AGCTGCAGATTCC-MGB | 250 | ||
| capB | F-primer | TAAGCCTGCGTTCTTCGTAAATG | 600 | |
| R-primer | GTTCCCAAATACGTAATGTTGATGAG | 600 | ||
| Probe | FAM-TTGCAGCGAATGAT-MGB | 250 |
* LightCycler 2.0 excitation by blue LED light source with maximum emission of 470 nm. Detection channels: 530, 560, 610, 640, 640, 705.