Figure 3

Clinical validation of high-throughput solid-phase protein truncation test (HTS-PTT) on BRCA1/2 exons 11 for inherited breast cancer susceptibility. One hundred blood-derived clinical genomic DNA samples were analyzed by HTS-PTT. (a) BRCA1 covered in two overlapping segments. (b) BRCA2 covered in three overlapping segments. (c) Standard curve of different protein inputs into the HTS-PTT enzyme-linked immunosorbent assay (ELISA) using serial dilutions of known wild-type samples. % C/N is the C- to N-terminal signal ratio of a given sample normalized against known wild-type controls. The green bars and lines indicate the designated threshold (3 standard deviations below the mean for the wild-type cohort), above which samples are scored as wild-type (blue bars) and below which samples are scored as mutant (red bars). Yellow bars indicate the predicted mutant % C/N based on the known wild-type standard curves (c). Black bars denote false-positives. The tick mark on the x-axis divides the 50 patients with known BRCA1/2 truncation mutations (left of tick mark) from the 50 wild-type patients (right of tick mark). BRCA1/BRCA2, breast and ovarian cancer susceptibility gene 1/2; RLU, relative light unit.