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 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.
Sotlar
K
K
Institute for Pathology, University of Tübingen, Tübingen, Germany. klstlar@med.uni-tuebingen.de
Selinka
H C
HC
Menton
M
M
Kandolf
R
R
Bültmann
B
B
D015139:Blotting, Southern; D002584:Cervix Uteri / Q000821:virology*; D004279:DNA, Viral / Q000302:isolation & purification; D004848:Epithelium / Q000821:virology; D005260:Female; D006801:Humans; D027383:Papillomaviridae / Q000145:classification / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000175:diagnosis* / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000302:isolation & purification; D012367:RNA, Viral / Q000302:isolation & purification; D012194:RNA-Directed DNA Polymerase; D014412:Tumor Virus Infections / Q000175:diagnosis* / Q000821:virology; D002583:Uterine Cervical Neoplasms / Q000175:diagnosis / Q000821:virology*; D002578:Uterine Cervical Dysplasia / Q000175:diagnosis / Q000821:virology*
D004279:DNA, Viral; D012333:RNA, Messenger; D012367:RNA, Viral; D012194:RNA-Directed DNA Polymerase