Table 2.
Selection of HRG primer and amount of RNA input for the Qiagen kit.a
| RNA origin |
Ct values |
||||||||
|---|---|---|---|---|---|---|---|---|---|
|
HRG primer/probe Bb |
HRG primer/probe Ab |
HRG primer probe Cb |
|||||||
| 20 ng | 50 ng | 100 ng | 20 ng | 50 ng | 100 ng | 20 ng | 50 ng | 100 ng | |
| FFPE NSCLC | |||||||||
| 1173692B | N/A | N/A | N/A | N/A | 36.7 | 36.6 | 38.2 | 37.3 | 35.4 |
| 1173127B | N/A | N/A | N/A | N/A | 38.6 | 37.7 | 38.0 | 36.5 | 37.0 |
| 1173740B | N/A | N/A | N/A | N/A | N/A | 38.7 | 39.0 | 38.2 | 37.3 |
| 2YHODAZN | 39.8 | N/A | 39.4 | 39.8 | 37.2 | 35.2 | 36.9 | 35.6 | 35.1 |
| AF8ALAN9 | 39.8 | 38.8 | 38.2 | 39.8 | 34.5 | 33.3 | 34.8 | 33.7 | 32.5 |
| QISIZA1M | 36.5 | 35.0 | 34.3 | 36.5 | 32.6 | 31.3 | 34.4 | 33.4 | 31.8 |
| TRQR7A81 | 32.1 | 31.5 | 30.0 | 32.1 | 28.4 | 27.3 | 29.4 | 28.3 | 26.8 |
| Cell linesc | |||||||||
| Fresh HRG+ | 29.1 | 29.0 | 27.2 | 27.3 | 26.9 | 25.4 | 26.9 | 26.4 | 24.5 |
| FFPE HRG+ | 36.7 | 35.4 | 34.3 | 36.7 | 31.2 | 30.3 | 32.1 | 30.9 | 29.3 |
| Fresh HRG(−) | N/A | N/A | N/A | N/A | N/A | 39.4 | 38.8 | 37.3 | 38.2 |
| FFPE HRG(−) | N/A | N/A | N/A | N/A | N/A | 38.8 | N/A | 38.8 | 37.1 |
Abbreviations: Ct, cycle threshold; FFPE, formalin-fixed paraffin-embedded; HRG, heregulin; N/A, not amplifiable; NSCLC, non–small cell lung cancer.
a
Mean Ct values for each assay for the 3 input amounts using Qiagen kit–extracted RNA.
b
Primer/probe A = Hs00247620_m1, B = Hs01108479_m1, and C = Hs01101537_m1.
c
HRG-positive cell line = A549 and HRG-negative cell line = T47D.