Abstract Objective.To evaluate whether the telomerase activity in CIN lesions can be affected by a chemical solution of acetic acid, which is required to apply to the cervix prior to colposcopy-directed biopsy. Materials and methods.Thirty-five patients with histologically confirmed high-grade squamous intraepithelial lesions of the cervix entered the study. Two specimens were collected from each patient, one before and one after the cervix was swabbed with 5% acetic acid. The standard telomeric repeat assay protocol (TRAP) was used to examine telomerase activity in these fresh frozen tissue samples. Normal cervical tissues from 10 control individuals were also examined for the presence of telomerase activity. A total of 80 specimens was analyzed. Results.Telomerase activity was detectable in 27 of 35 (77.1%) fresh tissue samples, 15 of 35 (42.9%) tissue samples swabbed with 5% acetic acid, and 0 of 10 (0%) normal cervical tissue samples, respectively. Twelve samples became telomerase negative after 5% acetic acid applied. Among the 15 telomerase-positive tissue samples swabbed with 5% acetic acid, 12 had relative weak telomerase activity compared to corresponding fresh tissue samples, the other 3 remained the same. Therefore, it is concluded that telomerase activity was affected by 5% acetic acid in 24 of 27 (88.9%) samples. Telomerase activity in HeLa cell line was also inhibited by 5% acetic acid. Conclusion.We reported a relative high percentage of telomerase expression in high-grade CIN lesions when compared with previous reports. If detection of telomerase activity is to become a tool for diagnosis and prognosis of cervical neoplasias, applying acetic acid prior to colposcopy-directed biopsy that is submitted for telomerase assay should be avoided in order to increase the detection rate.