TP53 and hepatocellular carcinoma: As this is the first study to validate the proposed system to specifically aiming to analyze c.747G > T hotspot mutations in TP53 gene of HCC, a concentration range of 2–200 fM target ctDNA concentrations were selected, aiming to fall into the reported plasma ctDNA range of 9.8 to 487.1 ng/mL, and corresponding EC responses were examined.