This study aimed to assess the diagnostic and prognostic value of four biochemical markers-PAX9, HK10, TBX2, and CYFRA21-1-in ovarian carcinoma (OC). We conducted a prospective diagnostic study from December 2020 to December 2021, enrolling 88 patients with ovarian carcinoma, 90 with benign ovarian lesions, and 92 healthy controls. Blood levels of HK10 and CYFRA21-1 were measured, while tissue expressions of PAX9 and TBX2 were analyzed. We used logistic regression and ROC curve analysis to evaluate the diagnostic performance of these markers. The OC group showed significantly higher positive expression rates of PAX9 and TBX2 compared to the benign group (65.91% vs. 4.44% and 26.14% vs. 3.33%, respectively). Additionally, HK10 and CYFRA21-1 levels were significantly elevated in the OC group compared to both the benign and healthy groups (P < 0.05). The expressions of all four markers were closely associated with clinical parameters such as tumor diameter, FIGO stage, degree of differentiation, and lymph node metastasis (P < 0.05). The sensitivity and specificity value of CA125, HK10, CYFRA21-1, PAX9, and TBX2 in the diagnosis of ovarian cancer were 85.09% and 80.09%, 65.87% and 61.67%, 81.25% and 79.59%, 75.32% and 74.49%, and 61.46% and 69.39%, respectively. The sensitivity and specificity of the combined biomarkers diagnosis are respectively 87.59% and 84.69%. The combined assessment of PAX9, HK10, TBX2, and CYFRA21-1 provides significant diagnostic and prognostic value in ovarian carcinoma, with HK10 showing particularly high sensitivity. These markers could play a crucial role in the early detection and management of ovarian carcinoma.