Table 3. Results for castPCR and Therascreen for each case included in the study.
| No. | TheraScreen Mutation Ct | TheraScreen DeltaCt | Therascreen Mutation | castPCR Mutation Ct | castPCR Delta Ct | castPCR Mutation |
|---|---|---|---|---|---|---|
| P1 | N/A | N/A | WT | N/A | N/A | WT |
| P2 | N/A | N/A | WT | 33.5 | 2.5 | BRAF c.1799T>A |
| P3 | 28.4 | 0.5 | KRAS c.35G>C | N/A | N/A | WT |
| P4 | 31.9 | 0.65 | KRAS c.35G>T | 33.1 | 3.0 | KRAS c.35G>T |
| P5 | 32.1 | 0.8 | KRAS c.35G>T | 35.0 | 4.3 | KRAS c.35G>T |
| P6 | 30.7 | 1.35 | KRAS c.35G>T | 28.0 | 2.3 | KRAS c.35G>T |
| P7 | 34.6 | 3.69 | KRAS c.35G>A | 32.5 | 3.5 | KRAS c.35G>A |
| P8 | N/A | N/A | WT | N/A | N/A | WT |
| P9 | N/A | N/A | WT | N/A | N/A | WT |
| P10 | N/A | N/A | WT | 26.2 | 0.6 | BRAF c.1799T>A |
| P11 | 34 | 4.12 | KRAS c.38G>A | 34.7, 35.3 | 5.4, 9.0 | KRAS c.38G>A, KRAS c.182A>G |
| P12 | 31.5 | 3.25 | KRAS c.34G>T | 28.0 | 3.1 | KRAS c.34G>T |
| P13 | N/A | N/A | WT | N/A | N/A | WT |
| P14 | N/A | N/A | WT | N/A | N/A | WT |
| P15 | N/A | N/A | WT | N/A | N/A | WT |
| P16 | 32.3 | 4.04 | KRAS c.38G>A | 29.3 | 4.0 | KRAS c.38G>A |
| P17 | 35.1 | 1.02 | KRAS c.35G>T | 33.5 | 1.8 | KRAS c.35G>T |
| P18 | N/A | N/A | WT | N/A | N/A | WT |
| P19 | 29.1 | 1.85 | KRAS c.35G>T | 27.1 | 2.8 | KRAS c.35G>T |
| P20 | N/A | N/A | WT | N/A | N/A | WT |
| P21 | 32.1 | 2.91 | KRAS c.34G>T | 31.3 | 4.2 | KRAS c.34G>T |
| P22 | 33.3 | 3.59 | KRAS c.35G>A | 33.0 | 3.9 | KRAS c.35G>A |
| P23 | 35.9 | 4.14 | KRAS c.38G>A | 33.2 | 3.1 | KRAS c.38G>A |
| P24 | N/A | N/A | WT | N/A | N/A | WT |
| P25 | 36.7 | 7.58 | KRAS c.35G>C | N/A | N/A | WT |
| P26 | N/A | N/A | WT | N/A | N/A | WT |
| P27 | N/A | N/A | WT | N/A | N/A | WT |
| P28 | N/A | N/A | WT | N/A | N/A | WT |
| P29 | 34 | 2.75 | KRAS c.35G>A | 33.4 | 3.6 | KRAS c.35G>A |
| P30 | N/A | N/A | WT | N/A | N/A | WT |
| P31 | 37.6 | 3.87 | KRAS c.38G>A | 33.0 | 4.9 | KRAS c.38G>A |
| P32 | 34.6 | 3.06 | KRAS c.35G>A | 38.5 | 7.7 | KRAS c.35G>A |
| P33 | N/A | N/A | WT | 27.4 | 1.1 | BRAF c.1799T>A |
| P34 | N/A | N/A | WT | N/A | N/A | WT |
| P35 | 31.6 | 3.88 | KRAS c.34G>A | 29.5 | 2.3 | KRAS c.34G>A |
| P36 | N/A | N/A | WT | 37.3 | 8.6 | BRAF c.1799T>A |
| P37 | N/A | N/A | WT | 29.0 | 2.0 | BRAF c.1799T>A |
| P38 | N/A | N/A | WT | N/A | N/A | WT |
| P39 | N/A | N/A | WT | 36.0 | 9.4 | BRAF c.1799T>A |
| P40 | N/A | N/A | WT | N/A | N/A | WT |
| P41 | N/A | N/A | WT | N/A | N/A | WT |
| P42 | N/A | N/A | WT | N/A | N/A | WT |
| P43 | 33.2 | 3.56 | KRAS c.38G>A | 36.9 | 6.5 | KRAS c.38G>A |
| P44 | N/A | N/A | WT | N/A | N/A | WT |
| P45 | 28.9 | 1.1 | KRAS c.35G>T | 28.7 | 2.2 | KRAS c.35G>T |
| P46 | N/A | N/A | WT | N/A | N/A | WT |
| P47 | 32.7 | 6.38 | KRAS c.34G>T | 30.4 | 4.2 | KRAS c.34G>T |
| P48 | N/A | N/A | WT | N/A | N/A | WT |
| P49 | 32.4 | 6.97 | KRAS c.35G>A | 28.2 | 2.7 | KRAS c.35G>A |
| P50 | N/A | N/A | WT | N/A | N/A | WT |
| P51 | N/A | N/A | WT | N/A | N/A | WT |
| P52 | N/A | N/A | WT | 39.4 | 9.4 | KRAS c.34G>A |
| P53 | 32 | 4.35 | KRAS c.35G>A | 30.9 | 3.6 | KRAS c.35G>A |
| P54 | N/A | N/A | WT | N/A | N/A | WT |
| P55 | N/A | N/A | WT | N/A | N/A | WT |
| P56 | 31.4 | 3.21 | KRAS c.34G>T | 30.4 | 3.7 | KRAS c.34G>T |
| P57 | N/A | N/A | WT | N/A | N/A | WT |
| P58 | 32.5 | 4.04 | KRAS c.35G>A | 32.8 | 4.3 | KRAS c.35G>A |
| P59 | 30 | 3.12 | KRAS c.35G>T | 29.6 | 4.4 | KRAS c.35G>T |
| P60 | N/A | N/A | WT | N/A | N/A | WT |
| P61 | N/A | N/A | WT | N/A | N/A | WT |
| P62 | 30.1 | 2.73 | KRAS c.34G>C | 27.7, | 3.89, | KRAS c.34G>C |
| P63 | N/A | N/A | WT | N/A | N/A | WT |
| P64 | N/A | N/A | WT | N/A | N/A | WT |
| P65 | N/A | N/A | WT | N/A | N/A | WT |
| P66 | N/A | N/A | WT | 27.3 | 1.6 | BRAF c.1799T>A |
| P67 | 31.6 | 3.97 | KRAS c.35G>A | 28.6 | 3.6 | KRAS c.35G>A |
| P68 | 32.2 | 4.26 | KRAS c.35G>A | 30.2 | 4.4 | KRAS c.35G>A |
| P69 | N/A | N/A | WT | N/A | N/A | WT |
| P70 | N/A | N/A | WT | N/A | N/A | WT |
| P71 | N/A | N/A | WT | 26.9 | 1.4 | BRAF c.1799T>A |
| P72 | N/A | N/A | WT | N/A | N/A | WT |
| P73 | N/A | N/A | WT | N/A | N/A | WT |
| P74 | 29.5 | 2.16 | KRAS c.34G>C | 28.1 | 3.2 | KRAS c.34G>C |
| P75 | 29.7 | 1.68 | KRAS c.35G>T | 28.5 | 2.5 | KRAS c.35G>T |
| P76 | 30.5 | 2.02 | KRAS c.35G>T | 29.8 | 4.5 | KRAS c.35G>T |
| P77 | N/A | N/A | WT | N/A | N/A | WT |
| P78 | N/A | N/A | WT | N/A | N/A | WT |
| P79 | N/A | N/A | WT | N/A | N/A | WT |
| P80 | N/A | N/A | WT | N/A | N/A | WT |
| P81 | 34.1 | 2.33 | KRAS c.34G>C | 33.1 | 4.0 | KRAS c.34G>C |
| P82 | N/A | N/A | WT | N/A | N/A | WT |
| P83 | N/A | N/A | WT | N/A | N/A | WT |
| P84 | N/A | N/A | WT | 27.1 | 1.6 | BRAF c.1799T>A |
| P85 | 36.6 | 4.58 | KRAS c.38G>A | 34.3 | 5.1 | KRAS c.38G>A |
| N/A | N/A | WT | 28.8 | 2.7 | BRAF c.1799T>A | |
| P86 | 30.8 | 2.3 | KRAS c.35G>T | 30.0, 35.1 | 4.1, 8.9 | KRAS c.35G>T, BRAF c.1799T>A |
| P87 | N/A | N/A | WT | N/A | N/A | WT |
| P88 | N/A | N/A | WT | N/A | N/A | WT |
| P89 | N/A | N/A | WT | N/A | N/A | WT |
| N/A | N/A | WT | N/A | N/A | WT | |
| P90 | N/A | N/A | WT | 27.0 | 1.5 | BRAF c.1799T>A |
| W1 | 27.85 | 2.14 | KRAS c.35G>C | 30.5 | 3.4 | KRAS c.35G>C |
| W2 | 30.53 | 2.26 | KRAS c.35G>A | 29.7 | 2.6 | KRAS c.35G>A |
| W3 | 28.57 | 2.86 | KRAS c.34G>T | 31.4 | 4.0 | KRAS c.34G>T |
| W4 | N/A | N/A | WT | N/A | N/A | WT |
| W5 | N/A | N/A | WT | 28.7 | 1.3 | BRAF c.1799T>A |
| W7 | N/A | N/A | WT | N/A | N/A | WT |
| W8 | N/A | N/A | WT | N/A | N/A | WT |
Ct values for each reference gene are given with the ΔCt. An analysis matrix is used to determine the assay result (mutant or wild-type) according to thresholds defined by the manufacturer. There were three discrepant results (shown in bold): P3 showed KRAS c.35G>C by both methods on retesting, while P25 and P52 were both wild-type. Two cases (P11 and P86) showed amplification for more than one mutation. N/A, not applicable; WT, wild-type.