Figure 2
Sensitivity of RT-qPCR using TLDA platform. Expression levels of 93 candidate genes were analyzed using cDNA generated from total RNA isolated from peripheral blood samples from a healthy female donor and the same blood spiked with 4, 40 and 400 T47-D tumor cells after cell enrichment. RNA was pre-amplified using the TargetAmp™1-Round aRNA Amplification Kit. Average Ct values obtained from RT-qPCR amplification of CCNE2, DKFZp762E1312, EMP2, MAL2, PPIC, and SLC6A8 transcripts using the TLDA format are shown. MAL2 and CCNE2 gene expression was below the detection limit of RT-qPCR in the unspiked blood. The detection sensitivity of the respective marker gene was estimated to be 40 and 400 tumor cells per 15 ml whole blood.