155 Detection of non-metastatic non-small cell lung cancer in urine differences between sex (P = .43) and age (P = .64). The proportions of never, former, and current smokers significantly differed between NSCLC patients and controls (P < 0.001). Two urine samples of NSCLC patients had insufficient quality (ACTB Ct ≥ 32) and were therefore excluded from further analysis, resulting in a study population of 44 patients for present methylation analysis. All control samples met the DNA quality criteria. Clinical characteristics of NSCLC patients and controls with valid qMSP results are depicted in Table 1. Table 1: Baseline characteristics of NSCLC patients (n=44) and controls (n=50). NSCLC Controls P Age Median (IQR) 66 (62-72) 66 (59-80) .64 Sex n % n % Male 21 47.7 28 56.0 .43 Female 23 52.3 22 44.0 Stage* n % 1 28 63.6 NA 2 4 9.1 3 12 27.3 4 0 0,0 Histology n % LUAD 27 61.4 NA LUSC 16 36.4 NOS 1 2.3 Smoking n % n % Never smokers 3 6,8 30 60,0 <0.001 Former smokers (stopped >1yr) 12 27,3 16 32,0 Current smokers (active or stopped <1yr) 28 63,6 4 8,0 Unknown 1 2,3 0 0,0 *Staging was according the 8th edition of the TNM criteria. IQR = interquartile range, LUAD = lung adenocarcinoma, LUSC = lung squamous cell carcinoma, NOS = carcinoma not otherwise specified, NSCLC = non-small cell lung cancer. DNA methylation levels in pre-operative urine samples DNA methylation levels of markers CDO1, SOX17, and TAC1 relative to ACTB were determined in the urine supernatant of 44 surgical NSCLC patients and 50 controls. Methylation levels of CDO1 and SOX17 were significantly higher in NSCLC patients as compared to controls (P = .016 and P < .001, respectively), while TAC1 did not show significant differences between groups (P = .347, Figure 1). As the proportion of smokers differs between the NSCLC 6
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