Table 9.
Real-time PCR results of 14-plex assay
| Organism | Strain | Agent | F. tularensis assays | Y. pestis assays | Burkholderia assays | B. anthracis assays | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| F. tularensis | 239 | A | 30.63 | 32.97 | TN | TN | – | – | – | – | – | – | – | – | – | – |
| 240 | B | 26.05 | TN | 28.75 | TN | – | – | – | – | – | – | – | – | – | – | |
| 241 | C | 29.80 | TN | TN | 30.97 | – | – | – | – | – | – | – | – | – | – | |
| Y. pestis | 113 | D | – | – | – | – | 24.86 | 23.04 | TN | 30.28 | – | – | – | – | – | – |
| 114 | E | – | – | – | – | 27.75 | TN | 29.71 | 31.04 | – | – | – | – | – | – | |
| B. mallei | 164 | F | – | – | – | – | – | – | – | – | 32.81 | TN | TN | – | – | – |
| B. pseudomallei | 197 | G | – | – | – | – | – | – | – | – | TN | 32.60 | 32.14 | – | – | – |
| B. anthracis | 708 | H | – | – | – | – | – | – | – | – | – | – | – | 27.07 | 28.45 | 28.97 |
| PCR Assay Number | 23 | 28 | 29 | 30 | 9 | 11 | 14 | 15 | 49 | 50 | 65 | 01 | 04 | 07 | ||
| Probe Dye Channel | FAM | VIC | NED | CY5 | FAM | VIC | NED | CY5 | FAM | VIC | NED | FAM | VIC | NED | ||
Real-time PCR results using a mixed assay of all 14 sets of primers and probes tested on individual agents. Each sample contained a single agent, extracted in singlet and analyzed by PCR. Agent concentration are all at 2.5E+ 05 CFU/mL. Each PCR reaction employed all 14 primer/probe sets. Data shows that only agent specific primer/probe sets detected with no FP or FN. Values in the cells indicate the Ct value of an observed positive result (Ct < 40) where a positive result was expected. A minus sign (−) indicates the assay-organism combination was not tested. Cells containing TN indicate an observed negative or undetected result (Ct ≥ 40) where a positive result was not expected