This T4-assisted system enables methylation analysis of RXFP3-L1, RXFP3-L2, ZNF671, PAX1, and SOX1 for cervical cancer (CESC) evaluation with only 1.5 ng of DNA input; furthermore, lower input is sufficient for assessing the hypermethylated PAX1 gene.<h4>Results</h4>When applied to 24 blood samples from CESC patients and 21 matched tissues, the modified MSRE-qPCR method achieved 87.5% sensitivity and 90.3% specificity using 1.5ng of DNA, outperforming MethyLight without requiring more sensitive quantitative techniques. Here, PAX1 is linked to cervical carcinoma.