Increased levels of HPV16 E6*I transcripts in high-grade cervical cytology and histology (CIN II+) detected by rapid real-time RT-PCR amplification

Cytopathology. 2007 Oct;18(5):290-9. doi: 10.1111/j.1365-2303.2007.00481.x. Epub 2007 Jul 27.


Objective: As cervical dysplasia may regress to normal cytology or progress to cervical carcinoma, it would be valuable to have a diagnostic tool to help decide whether therapeutic conization should be performed.

Methods: Cervical samples of 301 HPV16 positive women were collected in RNAlater reagent to prevent RNA degradation. Relative levels of HPV16 DNA and HPV16 E6*I mRNA in the samples were determined using real-time polymerase chain reaction. Findings were correlated with histological diagnoses and cytological follow-up.

Results: HPV16 E6*I mRNA levels were significantly higher in women with cytologically diagnosed severe cervical dysplasia (mean normalized ratio = 0.25) than in those with mild-to-moderate dysplasia (mean norm. ratio = 0.12), atypical squamous cells of uncertain origin (mean norm. ratio = 0.071) or normal cytology (mean norm. ratio = 0.034). Viral DNA levels were not significantly different between severe and mild-to-moderate dysplasia (mean norm. ratios, 55.8 and 63.5, respectively). The PPV for a histological diagnosis of severe cervical dysplasia [cervical intraepithelial neoplasia (CIN) II+] increased with the amounts of E6*I mRNA to more than 90% whereas the sensitivity decreased. The absence of HPV16 E6*I transcripts as well as HPV16 DNA considerably increased the negative predictive value and the specificity. However, low concentrations (or complete absence) of E6*I mRNA did not preclude a CIN II+ diagnosis.

Conclusions: Although the sensitivity is low, high levels of HPV16 E6*I mRNA are indicative of CIN II+ in cytologically diagnosed cervical dysplasia of individual patients. Thus, quantification of HPV16 E6*I mRNA could be helpful in managing follow-up and treatment in a subset of HPV16 positive women.

MeSH terms

  • Adolescent
  • Adult
  • Biomarkers, Tumor / analysis*
  • Cervical Intraepithelial Neoplasia / pathology*
  • Cervical Intraepithelial Neoplasia / virology*
  • DNA, Viral / analysis
  • Female
  • Humans
  • Middle Aged
  • Oncogene Proteins, Viral* / genetics
  • Papillomavirus Infections / complications
  • Precancerous Conditions / pathology
  • Precancerous Conditions / virology
  • Prognosis
  • RNA, Messenger / analysis
  • RNA, Viral / analysis
  • Repressor Proteins* / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sensitivity and Specificity
  • Tumor Virus Infections / complications
  • Uterine Cervical Dysplasia / pathology
  • Uterine Cervical Dysplasia / virology
  • Uterine Cervical Neoplasms / pathology*
  • Uterine Cervical Neoplasms / virology*


  • Biomarkers, Tumor
  • DNA, Viral
  • E6 protein, Human papillomavirus type 16
  • Oncogene Proteins, Viral
  • RNA, Messenger
  • RNA, Viral
  • Repressor Proteins