Objective: To define the relationship between the number of cervi-cal colposcopic biopsies performed on a patient and the diagnosis of each grade of cervical intraepithelial neoplasia (CIN). Methods: Pa-tients who underwent a colposcopy and biopsy between January and June 2018 in an Italian second-level check-point for cervical cancer screening were prospectively enrolled in the study. Cervical punch biopsies were performed on abnormal acetowhite areas that were identified by colposcopy and endocervical sampling was performed if needed. The number of cervical biopsies per patient was recorded along with the following parameters: type of transforming zone, col-poscopic grading, Pap smear result, the patient's age, and endocervi-cal sampling. All parameters were included in multivariable models. The dependent variable was a diagnosis of CIN-0/1, CIN-2, or CIN-3. Results: Independently of other variables, a Pap test result of atypical squamous cells—cannot be excluded H-SIL (ASC-H), atypical glan-dular cells, not otherwise specified (AGC-NOS), or high grade squa-mous intraepithelial lesion (H-SIL) is associated with reduced odds of a CIN-0 or CIN-1 diagnosis. More than one cervical biopsy per pa-tient is associated with reduced odds of a CIN-0 or CIN-1 diagnosis whereas three or four biopsies is associated with increased odds of a CIN-2 diagnosis. A Pap test result of HSIL, ASC-H, or AGC-NOS is the only variable that increased the odds of a CIN-3 diagnosis. Dis-cussion: A greater number of cervical biopsies performed on a patient increases the likelihood of diagnosing a CIN-2 but has no effect on the diagnoses of CIN-0/1 or CIN-3.