Human Papilloma Virus Analysis of HPV FISH patterns in low and high grade Cervical Intraepithelial Neoplasia

  • V.M.M. van Meegen


Human Papilloma Virus is the most common sexually transmitted infection, with anestimated 80% of sexually active men and women acquiring an infection at some pointin their lifetime. 10-20% of infected individuals can not clear this infection effectivelyand consequentially are at risk for progression of Cervical Intraepithelial Neoplasia(CIN) to cancer. Presence of HPV can be determined using PCR and/or (Fluorescence) InSitu Hybridization. The aim of the performed experiments was to determine the generalFISH patterns that are specifically linked to low grade and high grade CIN lesions and toinvestigate whether or not these patterns could be used to grade these lesions.12 formalin fixed and paraffin embedded sections from one patient and 30 formalin fixedand paraffin embedded sections from different patients where used to perform a FISHprocedure and to analyze the general FISH hybridization pattern for CIN 1,2 and 3. For theanalysis 3 distinct patterns for the physical status of the virus were determined: episomal,integrated and mixed pattern. Also the presence of replication, load and the ratio betweenbasal load and superficial load was analyzed to determine the general pattern.Results show that load and physical status of the virus are not associated with the severityof the lesions. High loads are present in both high and low grade lesions. Also physicalstatus of the virus is not different for the sections, episomal and mixed patterns are foundin low and high grades. Only integrated pattern is a marker for severity, as this is only foundin CIN 3. Presence of replication is most common in CIN 1, this might contribute to correctgrading. The ratio between the load in the Basal layer and the load in the superficial layeris the most informative discriminant of severity: <0.5 for CIN 1, 0.5<load<1 for CIN 2 and 1for CIN 3. Based on these results it is possible to classify the severity of the lesion


Stanley M; HPV-immune response to infection and vaccination; Infectious agents and cancer;2010;5;19

Burd EM; Human Papillomavirus and Cervical Cancer; Clinical Microbiology Reviews; 2003;16;1;1-17

Muñoz N;Castellsagué X et al; Chapter 1: HPV in the etiology of human cancer; Vaccine; 2006;24

Molijn A; Kleter B et al; Molecular diagnosis of human papillomavirus (HPV) infections; journal of clinical virology;2005;S43-S51

Banks L; Pim D and Thomas M; Human tumour viruses and the deregulation of cell polarity in cancer; Nature Reviews Cancer, 2012;12;877-886 Solomon D; Davey D; Kurman R et al; The 2001 Bethesa System: terminology for reporting results of cervical cytology; The Journal of the American Medical Association;2002;287;16;2114-2119

Villa LL; Denny L; Methods for detection of HPV infection and its clinical utility; International Journal of Gynecology and Obstetrics;2006;94;S71-S80

Birmer P; Bachtiary B et al; signal amplified colorimetric in situ hybridization for assessment of human papillomarvirus infection in cervical lesions;Modern Pathology 2001;14;7;702-709

Fluorescence in Situ Hybridization; Nature Methods;2005;2;237-238

Speel EJ; Schutte B et al; The effect of avidin-biotin interactions in detection systems for in situ hybridization; the journal of histochemistry and cytochemistry;1992;40;1;135-141

Hopman AHN; Remaekers FCS; Current protocols in cytometry; John Wiley and Sons,Inc; 1998;8.5.1 -8.5.22

Evans MF; Mount SL et al; biotinyl-tyrammide based in situ hybridization signal patterns distinguish human papillomavirus type and grade of cervical intraepithelial neoplasia;modern pathology;2002;15;12;1339-1347

Cooper K; Herrington CS et al; Episomal and integrated human papillomavirus in cervical neoplasia shown by non-isotopic in situ hybridization; journal of clinical pathology;1991;44;990-996

Kristiansen E, Jenkins A et al; Coexistence of episomal and integrated HPV16 DNA in squamous cell carcinoma of the cervix; journal of clinical pathology;1994;47;253-256

Chatuverdi AK et al; Human papillomavirus infections with multiple types; pattern of coinfection and risk of cervical disease; The Journal of Infectious Diseases;2001;2011;203;910-920

Rousseau MC; Pereira JS et al; Cervical coinfection with human papillomavirus (HPV) types as a predictor of acquisitions and persistence of HPV infection; Journal of Infectious Diseases;2001;15;1508-1517

Feller L; Khammissa RAG et al; epithelial maturation and molecular biology of oral HPV; Infectious agents and cancer;2009;4;16