Liquid biopsy is a minimally invasive approach to detect early cancer in body fluids such as blood and urine 3, 4, 5. The detection and treatment of cancer when the disease is still at an early stage could save many lives, reduce morbidity, and relieve the burden of cancer on healthcare systems 1, 2. Therefore, the inclusion of 5-hydroxymethylcytosine in multianalyte testing could improve sensitivity for the detection of early-stage cancer. We confirm that 5-hydroxymethylcytosine can be used to detect CRC, even in early-stage disease. Expanding the classifier to include information about cell-free DNA fragment size and abundance across the genome led to gains in sensitivity (63% at 95% specificity), with similar overall performance (AUC 91% 95% CI ). The classifier could detect CRC even when the proportion of tumor DNA in blood was undetectable by other methods. Performance was similar for early stage 1 (AUC 89% 95% CI ) and late stage 4 CRC (AUC 94% 95% CI ). In an independent validation sample set, the classifier discriminated CRC samples from controls with an area under the receiver operating characteristic curve (AUC) of 90% (95% CI ). In a large covariate-controlled study of over two thousand individual blood samples, we created, tested and explored the properties of a 5-hydroxymethylcytosine-based classifier to detect colorectal cancer (CRC). The blood biomarker 5-hydroxymethylcytosine has been shown to discriminate cancer. Scientific Reports volume 12, Article number: 16566 ( 2022)Įarly detection of cancer will improve survival rates. Hydroxymethylation profile of cell-free DNA is a biomarker for early colorectal cancer
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