Background: Ovarian carcinoma (OC) has an unfavorable prognosis due to lack of screening and an asymptomatic course. New diagnostic methods are being sought to enable earlier diagnosis of this condition. The purpose of this study was to determine the diagnostic utility of collagenases (MMP-1, MMP-8 and MMP-13) in the diagnosis of OC compared to HE4 and CA125 and the ROMA. Methods: The study group consisted of 120 patients with OC, the control group: 70 patients with benign ovarian lesions (BLs) and 50 healthy women (HS). MMP-1, MMP-8 and MMP-13 were determined by ELISA and HE4 and CA125 by CMIA. Results: OC patients had higher levels of MMP-1 and MMP-13 compared to the BL and HS groups. MMP-1 (SE: 81.66%; SP: 94%; PPV: 97.02%; NPV: 68.11%; AUC: 0.9625) and MMP-13 (SE: 77.50%; SP: 94%; PPV: 96.875%; NPV: 63.51%; AUC: 0.917) showed similar or higher diagnostic values to routine markers (HE4: SE:85%; SP: 92%; PPV: 96.22%; NPV: 71.875%; AUC: 0.943; CA125: SE: 80%; SP: 98%; PPV: 98.96%; NPV: 67.12%; AUC: 0.909) and the ROMA (SE: 90.83%; SP: 94%; PPV: 97.32%; NPV: 81.03%; AUC: 0.955). Performing combined analyses of individual MMPs and MMPs with ROMA was associated with further increases in diagnostic parameters. Conclusions: MMP-1 and MMP-13 have shown preliminary potential as diagnostic markers and auxiliary markers to ROMA in biochemical diagnosis of OC.