Non-invasive diagnosis of early-stage lung cancer using high-throughput targeted DNA methylation sequencing of circulating tumor DNA (ctDNA).

In order to assess assay sensitivity, we first selected informative CpG regions based on the following criteria: (1) the percentage of co-methylated reads (co-methylated reads / all mapped reads with at least 3 CpGs) is required to be greater than 7.5% from the undiluted tumor gDNA sample; (2) the percentage of co-methylated reads at dilution 1% is required to be greater than the percentage of co-methylation at background using WBC gDNA; (3) at least one replicate of dilutions from 1% to 100% conform to an increasing trend.

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