DETECTION OF HUMAN-PAPILLOMAVIRUS TYPE-16 E6 E7 ONCOGENE TRANSCRIPTS IN DYSPLASTIC AND NONDYSPLASTIC CERVICAL SCRAPES BY NESTED RT-PCR/

Infections with high-risk human papillomaviruses (e.g., HPV-16) play a n important role in the development of cervical intraepithelial neopla sia (CIN) and invasive cervical cancer (IC). Continued expression of t he viral E6 and E7 genes is believed to be a key factor for oncogenic transformation of infected cells. Two spliced transcripts of the E6/E7 oncogenes, termed E6I and E6*II, can be detected by reverse transcri ptase polymerase chain reaction (RT-PCR). To increase the sensitivity of detecting E6/E7 transcripts in cervical scrapes we took advantage o f a nested RT-PCR (nRT-PCR) protocol. In a series of 30 HPV-16-positiv e patients with histologic diagnoses ranging from nondysplastic epithe lium to IC, the application of nRT-PCR significantly improved the dete ction of E6/E7 transcripts compared to conventional RT-PCR. The preval ence of E6/E7 spliced transcripts correlated with lesion severity and the nRT-PCR protocol allowed detection of these transcripts even in no ndysplastic epithelium and CIN I lesions. Therefore, in epidemiologic follow-up studies, detection of E6/E7 transcripts by nRT-PCR should pr ove to be a useful diagnostic tool for risk evaluations regarding the development of CIN and its progression to cervical cancer, especially in high-risk HPV-type-infected patients with CIN 0 and CIN I. (C) 1998 Academic Press.