SHOX2 DNA Methylation Is a Biomarker for the Diagnosis of Lung Cancer in Plasma Christoph Kneip, PhD, Bernd Schmidt, MD, Anke Seegebarth, Sabine Weickmann,

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SHOX2 DNA Methylation Is a Biomarker for the Diagnosis of Lung Cancer in Plasma Christoph Kneip, PhD, Bernd Schmidt, MD, Anke Seegebarth, Sabine Weickmann, Michael Fleischhacker, PhD, Volker Liebenberg, MD, John K. Field, MD, PhD, Dimo Dietrich, PhD Journal of Thoracic Oncology Volume 6, Issue 10, Pages 1632-1638 (October 2011) DOI: 10.1097/JTO.0b013e318220ef9a Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 1 Analytical assay performance. Analytical performance of the quantitative real-time polymerase chain reaction assay for quantifying SHOX2 DNA methylation. Different amounts of methylated DNA (3–720 pg) were spiked into a background of 50,000 pg unmethylated DNA. Number of replicates: 12 (0–60 pg), 6 (180 and 360 pg), and 4 (720 pg), respectively. Journal of Thoracic Oncology 2011 6, 1632-1638DOI: (10.1097/JTO.0b013e318220ef9a) Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 2 Results of the training study. Plasma from 20 individuals without cancer (controls) and 20 late stage (stage IV) patients with lung cancer (cases) was analyzed. Shown is a receiver operating characteristic plot (A) and the single SHOX2 methylation values (B). A methylation cutoff of 0.05% was used to discriminate between plasma from cases and controls (B). Journal of Thoracic Oncology 2011 6, 1632-1638DOI: (10.1097/JTO.0b013e318220ef9a) Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 3 Results of the testing study. Box plot displaying SHOX2 DNA methylation values measured in controls, cases, and in cases according to tumor stages. A methylation cutoff of 0.05% as determined in the training study was used to stratify patients into SHOX2 methylation-positive (>0.05%) and SHOX2 methylation-negative (<0.05%) groups. Journal of Thoracic Oncology 2011 6, 1632-1638DOI: (10.1097/JTO.0b013e318220ef9a) Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 4 Results of the clinical performance evaluation study. A, Receiver operating characteristic plot. B, Resulting sensitivity and specificity of the assay when using alternative cutoffs for patient stratification. Journal of Thoracic Oncology 2011 6, 1632-1638DOI: (10.1097/JTO.0b013e318220ef9a) Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions