Increase in viral load, viral integration, and gain of telomerase genes during uterine cervical carcinogenesis can be simultaneously assessed by the HPV 16/18 MLPA-assay

Am J Pathol. 2010 Oct;177(4):2022-33. doi: 10.2353/ajpath.2010.090901. Epub 2010 Sep 2.


Oncogenic human papillomavirus (HPV) infection is the most important risk factor in cervical carcinogenesis cases; high viral loads, viral integration into the host genome, and gain of the telomerase-related genes, TERT and TERC, are all factors associated with progression to cancer. A recently developed multiparameter HPV 16/18 multiplex ligation-dependent probe amplification (MLPA) assay, which allows the simultaneous assessment of these factors, was applied to a series of 67 normal and (pre)malignant frozen uterine cervical samples, as well as to 91 cytological preparations, to test the ability of the MLPA assay to identify high-risk lesions on the basis of these factors. Validation was performed using quantitative PCR, the PapilloCheck and fluorescence in situ hybridization. Only 5 out of 37 normal tissue samples or low-grade cervical lesions (ie, CIN1 and condyloma) showed either an HPV16 viral load higher than 25 copies per cell, viral integration, and/or gain of one of the telomerase-related genes, whereas for the high-grade cervical lesions, one or more of these risk factors was found in 25 of 30 cases. The HPV MLPA assay showed a sensitivity of 83% and a specificity of 86% in frozen cervical specimens. Furthermore, the feasibility of the MLPA assay was shown for cytological samples, where in 57% of high-grade squamous intraepithelial lesion cases, the high-risk factors were detected using this assay.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Carcinoma, Squamous Cell / diagnosis
  • Carcinoma, Squamous Cell / etiology
  • Cell Transformation, Neoplastic / genetics
  • Cell Transformation, Neoplastic / pathology
  • Cervical Intraepithelial Neoplasia / diagnosis
  • Cervical Intraepithelial Neoplasia / etiology
  • DNA, Viral / genetics
  • Feasibility Studies
  • Female
  • Human papillomavirus 16 / isolation & purification*
  • Human papillomavirus 18 / isolation & purification*
  • Humans
  • In Situ Hybridization, Fluorescence
  • Middle Aged
  • Nucleic Acid Amplification Techniques
  • Papillomavirus Infections / diagnosis*
  • Papillomavirus Infections / etiology
  • Polymerase Chain Reaction
  • Telomerase / genetics*
  • Telomerase / metabolism
  • Uterine Cervical Neoplasms / diagnosis*
  • Uterine Cervical Neoplasms / etiology
  • Uterus / metabolism
  • Uterus / pathology
  • Viral Load*
  • Virus Integration*
  • Young Adult


  • DNA, Viral
  • TERT protein, human
  • Telomerase