In the present study, we have for the first time demonstrated that the levels and frequencies of PAX1 and ZNF582 methylation are markedly higher in the tumor tissues compared to non-tumor tissues from ESCC patients, and methylation testing of these two genes has an excellent accuracy and great sensitivity and specificity to detect ESCC tumors, suggesting that PAX1 and ZNF582 methylation testing may be a promising biomarker for the detection of ESCC. Here, ZNF582 is linked to esophageal squamous cell carcinoma.