Objective: To evaluate whether E7 mRNA can predict the risk of progression in women with HPV16 infection. Design: prospective observational study Setting: Tertiary university hospital Population: A cohort of 139 women referred to colposcopy for an abnormal screening result fulfilling the following inclusion criteria: 1) a positive test result confirming HPV16 infection; 2) a biopsy sample with a histological diagnosis of absence of lesion or low-grade SIL/CIN grade1 (LSIL/CIN1); 3) no previous HPV vaccination; 4) no pregnancy; and 5) no previous cervical treatments; and 6) no immunosuppression. Methods: At the first visit all women underwent a cervical sample for liquid-based cytology, HPV testing and genotyping, and HPV16 E7 mRNA analysis and a colposcopy with at least one colposcopy-guided biopsy. Follow-up visits were scheduled every six months. In each control a liquid-based Pap smear, HPV testing, as well as a colposcopy examination with biopsy if necessary were performed. Main outocome measures: Histological diagnosis of HSIL/CIN2+ at any time during follow-up Results: E7 mRNA expression was positive in 55/127 (43.3%) women included in the study and seven (12.7%) progressed to HSIL/CIN2+. In contrast, only 1/72 (1.4%) women with no HPV16 E7 mRNA expression progressed (p=0.027). HPV16 E7 mRNA expression was associated with a 10-fold increased risk of progression (HR 10.0; 95%CI 1.2-81.4). Conclusions: HPV16 E7 mRNA could be useful for risk stratification of women with HPV16 infection in whom a HSIL/CIN2+ has been ruled out.