Table 1.

Step-wise validation results for a selected subset of 25 marker candidates derived from the discovery process and from the literature.¹

Gene symbol (HUGO)	Methylation array			Real-time PCR
Gene symbol (HUGO)	AUC, CRC vs all controls	AUC, CRC vs colon controls	PBL: median PMR, %	PBL: median PMR, %	Healthy colon: median PMR (90% range), %	CRC: median PMR, (90% range), %	AUC	Exclusion criteria
BCL6	0.73	0.78	25					Failed chip-selection criteria
DNAJC5	0.67	0.73	23
DUX2	0.75	0.70	17
ESTG020896²	0.64	0.63	9
MSH6³	0.58	0.64	0
ESTG2308609²	0.62	0.56	22
DLX5	0.84	0.83	0	PBL+				Failed PBL filter
NPBWR1 (GPR7)	0.73	0.82	0	PBL+
APBA2³	NT⁴	NT	NT	PBL+
SLC32A1	0.85	0.89	0	1.0
SLITRK1	0.86	0.85	1	0.3
KCTD12	0.67	0.77	0	0.0	0 (0–0)	0 (0–28)	0.53	Insufficient CRC methylation
ONECUT2	0.72	0.77	2	0.0	1 (0–1)	0 (0–2)	0.24
RNF4	0.79	0.84	0	0.0	0 (0–0)	0 (0–0)	0.52
SMAD7	0.68	0.71	0	0.0	0 (0–0)	0 (0–1)	0.59
TUSC3³	0.82	0.74	1	0.2	3 (1–10)	11 (1–32)	0.71	Failed LOD test
TMEFF2	0.90	0.94	0	0.1	4 (1–9)	30 (1–81)	0.81
ZDHHC22	0.88	0.87	9	0.0	0 (0–1)	3 (0–27)	0.78	HM assay failed
VTN	0.93	NT	1	0.0	2 (0–4)	20 (0–77)	0.81	Plasma positive
FOXL2	0.67	NT	0	0.0	NT	7 (0–38)	NT	Candidates for plasma testing
NGFR	0.83	0.87	0	0.0	0 (0–2)	3 (0–60)	0.69
ALX4	0.65	0.83	0	0.1	1 (0–2)	11 (0–58)	0.74
SIX6	0.77	0.85	0	0.0	0 (0–0)	16 (0–46)	0.83
EYA4	0.66	0.81	4	0.0	2 (0–4)	26 (0–76)	0.88
SEPT9	0.84	NT	0	0.0	2 (1–7)	26 (0–89)	0.87