OBJECTIVES To investigate feasibility and effectiveness of a cervical screening program with DNA tests as preliminary assay versus usual cytology protocols in Umbria Region. METHODS A large cohort of 35-64 aged women afferent to the unique regional laboratory was considered. The usual algorithm with cervical cytology as primary test was followed in January 2008–June 2010, whereas in August 2010–October 2011 high-risk human papillomavirus (HR-HPV) DNA test was used as primary screening. The cohorts were compared in terms of acceptance rate of invitation, cytological results, molecular results including HPV genotype, detection rate of histological lesions. RESULTS : A total of 31,228 women were invited: 21,249 were suggested to undergo classical cervical cytology screening, 9,979 HR-HPV DNA test as primary screening. A similar rate of adhesion (56.6% vs. 56.5%) was observed. Age-related differences were evidenced, with younger women (35-49) more prone to accept the invitation to HR-HPV DNA testing rather than usual cytology screening (61.6% vs. 55.5%; p<0.0001); analogously, uninvited younger women spontaneously requesting cervical screening were more prone to specifically request molecular than classical cytological testing (24.8% vs. 10.8%; p<0.0001). Among the 6,272 HR-HPV DNA testing women, 396 (6.4%) were positive, and, among them, 141 (36%) featured an altered cytology. All patients with altered cytology were suggested to undergo colposcopy and 106 out of 141 (75.1%) answered to the invitation. Among them, 89 (84%) featured abnormal histology with 48 (45.3%) CIN1 and 41 (38.7%) CIN2. If comparing the CIN2 detection rate within the two studied periods, it was almost doubled using the HR-HPV DNA than pap test as primary assay (0.64% vs. 0.37%; p=0.005). Finally, the implementation of the DNA test screening program did not increase total costs. CONCLUSIONS although with some limits, the introduction of HR-HPV DNA primary testing resulted feasible and effective, significantly increasing detection of severe lesions.