Background <p>Cervical cancer continues to be a major worldwide health concern. Diagnostic methods are limited, and therefore, more specific, non-invasive serum biomarkers are required for greater efficacy in detecting and monitoring the disease.</p> Objective <p>This research aims to investigate the diagnostic and prognostic utility of serum Cytokeratin 5 (CK5) and p63 as biomarkers for cervical cancer.</p> Material and methods <p>A case–control study was conducted with 50 women with newly diagnosed, histologically confirmed cervical cancer and 50 age-matched healthy controls. Serum levels of CK5 and p63 were quantified using enzyme-linked immunosorbent assay (ELISA).</p> Results <p>Serum levels of both CK5 (286 ± 94 vs. 77.5 ± 28.2 ng/L, p &lt; 0.001) and p63 (35.1 ± 10.6 vs. 7.5 ± 3.2 ng/mL, <i>p</i> &lt; 0.001) were significantly elevated in cervical cancer patients compared to controls. ROC analysis revealed excellent diagnostic accuracy, with an area under the curve (AUC) of 0.96 for CK5 and 0.97 for p63 both biomarkers being statistically significantly more in advanced-stage than in early-stage of the disease on <i>p</i> = 0.002 and <i>p</i> &lt; 0.0001, respectively, and independent predictors of advanced disease in multivariate analysis.</p> Conclusion <p>Serum CK5 and p63 had an excellent diagnostic efficiency for cervical cancer and correlated with disease progression, positioning them as promising non-invasive biomarkers for diagnosis and staging.</p>