Detection of human papillomavirus type 16 E6/E7 oncogene transcripts in dysplastic and nondysplastic cervical scrapes by nested RT-PCR

Gynecol Oncol. 1998 May;69(2):114-21. doi: 10.1006/gyno.1998.4994.

Abstract

Infections with high-risk human papillomaviruses (e.g., HPV-16) play an important role in the development of cervical intraepithelial neoplasia (CIN) and invasive cervical cancer (IC). Continued expression of the 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 E6*I and E6*II, can be detected by reverse transcriptase polymerase chain reaction (RT-PCR). To increase the sensitivity of detecting E6/E7 transcripts in cervical scrapes we took advantage of a nested RT-PCR (nRT-PCR) protocol. In a series of 30 HPV-16-positive patients with histologic diagnoses ranging from nondysplastic epithelium to IC, the application of nRT-PCR significantly improved the detection of E6/E7 transcripts compared to conventional RT-PCR. The prevalence of E6/E7 spliced transcripts correlated with lesion severity and the nRT-PCR protocol allowed detection of these transcripts even in nondysplastic epithelium and CIN I lesions. Therefore, in epidemiologic follow-up studies, detection of E6/E7 transcripts by nRT-PCR should prove 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Southern
  • Cervical Intraepithelial Neoplasia / diagnosis
  • Cervical Intraepithelial Neoplasia / virology*
  • Cervix Uteri / virology*
  • DNA, Viral / isolation & purification
  • Epithelium / virology
  • Female
  • Humans
  • Papillomaviridae / classification
  • Papillomaviridae / isolation & purification*
  • Papillomavirus Infections / diagnosis*
  • Papillomavirus Infections / virology
  • Polymerase Chain Reaction / methods
  • RNA, Messenger / isolation & purification
  • RNA, Viral / isolation & purification
  • RNA-Directed DNA Polymerase
  • Tumor Virus Infections / diagnosis*
  • Tumor Virus Infections / virology
  • Uterine Cervical Neoplasms / diagnosis
  • Uterine Cervical Neoplasms / virology*

Substances

  • DNA, Viral
  • RNA, Messenger
  • RNA, Viral
  • RNA-Directed DNA Polymerase