Detection of Colorectal Cancer Using a Simplified SEPT9 Gene Methylation Assay Is a Reliable Method for Opportunistic Screening  Dong Wu, Guangpeng Zhou, Peng Jin, Jiqing Zhu, Shijie Li, Qi Wu, Guiqi Wang, Jianqiu Sheng, Jianming Wang, Lele Song, Xiaoliang Han, Jiaming Qian  The Journal of Molecular Diagnostics  Volume 18, Issue 4, Pages 535-545 (July 2016) DOI: 10.1016/j.jmoldx.2016.02.005 Copyright © 2016 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 1 Recruitment and clinical assignment of patients for opportunistic screening. Patients (N = 1200) were recruited to obtain a statistically significant number of CRC cases. Ten-milliliter blood samples were obtained from all patients, and, in addition, FIT and CEA analysis could be performed on 71 and 235 CRC subjects, respectively. Because of incomplete history or loss of tracking, 169 patients were excluded from the trial such that the final enrollment comprised 1031 subjects. All subjects received colonoscopy to confirm diagnosis and were divided into six clinical classifications. CEA, carcinoembryonic antigen; CRC, colorectal cancer; FIT, fecal immunochemical test; GI, gastrointestinal; NED, no evidence of disease. The Journal of Molecular Diagnostics 2016 18, 535-545DOI: (10.1016/j.jmoldx.2016.02.005) Copyright © 2016 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 2 The receiver operating characteristic curve for the new SEPT9 assay in the case-control training study. The AUC equals 0.873. Cycle threshold values from no evidence of disease and colorectal cancer subjects were used in plotting the curve. AUC, area under curve. The Journal of Molecular Diagnostics 2016 18, 535-545DOI: (10.1016/j.jmoldx.2016.02.005) Copyright © 2016 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 3 The number and ratio of recruited patients in each disease classification. A: In the histogram the number of patients is shown for cancer stage and disease class. B: These data are represented as percentages in the pie chart. CRC, colorectal cancer; GI, gastrointestinal; NED, no evidence of disease; NS, not specified. The Journal of Molecular Diagnostics 2016 18, 535-545DOI: (10.1016/j.jmoldx.2016.02.005) Copyright © 2016 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 4 Positive detection rate for NED, and all stages of CRC. The SEPT9 assay detects 64.9% stage I, 72.7% stage II, 79.3% stage III, and 93.9% stage IV CRC, and the overall sensitivity was 76.6%. In contrast, only 4.1% NED subjects show positive results, indicating that the SEPT9 assay is a specific test for CRC detection. Late-stage CRC exhibits a higher positive detection rate than early-stage CRC, which is consistent with previous observations with Epi proColon 2.0 in the Chinese population.23 Data are presented as calculated positive detection rate with 95% CI. CRC, colorectal cancer; NED, no evidence of disease. The Journal of Molecular Diagnostics 2016 18, 535-545DOI: (10.1016/j.jmoldx.2016.02.005) Copyright © 2016 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 5 The receiver operating characteristic curve for the new SEPT9 assay in opportunistic screening. The area under curve equals 0.882. Cycle threshold values from 291 NED and 295 colorectal cancer subjects were used in plotting the curve. AUC, area under curve; NED, no evidence of disease. The Journal of Molecular Diagnostics 2016 18, 535-545DOI: (10.1016/j.jmoldx.2016.02.005) Copyright © 2016 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions