title: Low-risk human papillomavirus type 6 DNA load and integration in cervical samples from women with squamous intraepithelial lesions.
abstract: BACKGROUND
  The association between human papillomavirus (HPV) viral load and high-grade squamous intraepithelial lesion (HSIL) of the uterine cervix has been demonstrated for high-risk HPV-16 but has not been investigated for low-risk HPV types.
  
  
  OBJECTIVE
  To determine the association between the presence of low-grade SIL (LSIL) and viral load of low-risk HPV type 6.
  
  
  METHODS
  107 HPV-6-positive cytobrush samples collected from 90 women (67 without SIL, 11 with LSIL, 5 with HSIL, 6 with SIL of unknown grade and 1 with a smear with LSIL and a normal colposcopy) were analyzed for their content of HPV-6 DNA.
  
  
  METHODS
  HPV-6 E6 and E2 DNA loads were measured in the cytobrush samples with two real-time PCR assays. HPV-6 integration was confirmed with restriction-site PCR.
  
  
  RESULTS
  HPV-6 DNA in cervical samples ranged from 1.8 x 10(2) to 4.25 x 10(8) copies per microg of cellular DNA. HPV-6 E6 DNA loads were higher in women with LSIL, with or without co-infection with high-risk HPV types, than in women without SIL (p=0.03). HPV-6 loads greater than 8.76 x 10(6)copies per microgDNA, corresponding to the mean HPV-6 load measured in women with LSIL, were more frequently detected in these women with LSIL than in women without LSIL, controlling for age (odds ratio 13.5, 95% confidence interval 1.1-172.4). Although 10 samples generated HPV-6 E6/E2 ratios above 2, there was no evidence of integration of HPV-6 by restriction-site PCR.
  
  
  CONCLUSIONS
  Samples from women with LSIL contained higher HPV-6 loads than women without SIL. HPV-6 DNA was not integrated into cellular DNA in these samples.
authors.affiliation: Département de Microbiologie-Infectiologie et Centre de Recherche, Centre Hospitalier de l'Université de Montréal (CHUM), Université de Montréal, Québec, Canada. francois.coutlee@ssss.gouv.qc.ca
authors.forename: François
authors.initials: F
authors.lastname: Coutlée
authors.forename: Helen
authors.initials: H
authors.lastname: Trottier
authors.forename: Simon
authors.initials: S
authors.lastname: Gagnon
authors.forename: Anita
authors.initials: A
authors.lastname: Koushik
authors.forename: Harriet
authors.initials: H
authors.lastname: Richardson
authors.forename: Michel
authors.initials: M
authors.lastname: Roger
authors.forename: Alex S
authors.initials: AS
authors.lastname: Ferenczy
authors.forename: Eduardo L
authors.initials: EL
authors.lastname: Franco
mesh_terms: D004279:DNA, Viral / Q000235:genetics*; D005260:Female; D052163:Human papillomavirus 6 / Q000302:isolation & purification*; D006801:Humans; D018307:Neoplasms, Squamous Cell / Q000821:virology*; D009856:Oncogene Proteins, Viral / Q000235:genetics; D030361:Papillomavirus Infections / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000379:methods; D002583:Uterine Cervical Neoplasms / Q000821:virology*; D014626:Vaginal Smears; D016662:Virus Integration
chemical_list: D004279:DNA, Viral; D009856:Oncogene Proteins, Viral