Objectives: The purpose of this study was to evaluate the clinical significance of genomic amplification of the human telomerase RNA gene (TERC) for cervical cancer screening and explore whether it can serve as a biomarker to improve the specificity of human papillomavirus (HPV) DNA testing for cervical cancer screening. Methods: One hundred twenty women, including 20 cases of normal (control), 14 cases of cervical intraepithelial neoplasia grade I (CIN I), 35 cases of CIN II, 36 cases of CIN III, and 15 cases of squamous cervical cancer diagnosed by histopathologic evaluation, were subjected to cytopathologic examination, TERC detection by fluorescence in situ hybridization (FISH), and HPV DNA testing by Hybrid Capture II. Results: TERC amplification was significantly associated with cytopathologic diagnosis (P < 0.001) and histopathologic evaluation (P < 0.001). The positive rate of TERC gain was significantly higher in patients with CIN III or squamous cervical cancer than in patients with CIN I or those in the control group (P < 0.001). The specificity and positive predictive value of FISH for detecting CIN II or more severe cervical lesions (≥CIN II) were obviously higher than those of HPV DNA testing (97.1% vs 52.9%, 98.7% vs 83.8%). Conclusions: TERC amplification analyzed by FISH may serve as an adjunctive test to HPV DNA testing for improving the specificity and positive predictive value of cervical cancer screening. Copyright © 2010 by IGCS and ESGO.