title: DNA methylation analysis in self-sampled brush material as a triage test in hrHPV-positive women.
abstract: BACKGROUND
  Primary high-risk human papillomavirus (hrHPV) testing in cervical cancer screening shows relatively low specificity, which makes triage testing necessary. In this study, DNA methylation analysis was compared with cytology for triage testing in hrHPV-positive women. Moreover, feasibility of DNA methylation analysis directly on brush-based self-sampled specimens was assessed.
  
  
  METHODS
  Non-responding women from population-based screening were invited to self-collect a cervico-vaginal specimen for hrHPV testing; hrHPV-positive women were referred to a physician for triage liquid-based cytology. DNA methylation analysis was performed on 128 hrHPV-positive physician-collected triage samples and 50 matched brush self-samples with QMSP for C13ORF18, EPB41L3, JAM3 and TERT.
  
  
  RESULTS
  In physician-taken triage material, DNA methylation analysis of JAM3 showed the highest combined specificity (88%) and sensitivity (82%) for detection of CIN3+, whereas cytology showed a specificity of 48% and a sensitivity of 91%. Out of 39 women with abnormal cytology and normal histology (false-positive by cytology), 87% were negative for JAM3 and 90% for C13ORF18 methylation. Agreement between DNA methylation analysis performed directly on the matched self-sampled material and physician-taken samples was 88% for JAM3 (κ=0.75, P<0.001) and 90% for C13ORF18 (κ=0.77; P<0.001).
  
  
  CONCLUSIONS
  DNA methylation analysis as a triage test in hrHPV-positive women is an attractive alternative to cytology. Furthermore, DNA methylation is feasible directly on brush-based self-samplers and showed good correlation with matched physician-taken samples. Direct molecular triage on self-collected specimens could optimise the screening program, especially for non-responders, as this would eliminate the need for an additional physician-taken scraping for triage testing.
authors.affiliation: Department of Gynaecologic Oncology, Cancer Research Center, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands.
authors.forename: A
authors.initials: A
authors.lastname: Boers
authors.affiliation: Department of Obstetrics and Gynaecology, Radboud University Medical Center, Nijmegen, The Netherlands.
authors.forename: R P
authors.initials: RP
authors.lastname: Bosgraaf
authors.affiliation: Department of Gynaecologic Oncology, Cancer Research Center, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands.
authors.forename: R W
authors.initials: RW
authors.lastname: van Leeuwen
authors.affiliation: Department of Pathology, Cancer Research Center, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands.
authors.forename: E
authors.initials: E
authors.lastname: Schuuring
authors.affiliation: Department of Pathology, VU University Medical Center, Amsterdam, The Netherlands.
authors.forename: D A M
authors.initials: DA
authors.lastname: Heideman
authors.affiliation: Department of Obstetrics and Gynaecology, Radboud University Medical Center, Nijmegen, The Netherlands.
authors.forename: L F A G
authors.initials: LF
authors.lastname: Massuger
authors.affiliation: Department of Pathology, VU University Medical Center, Amsterdam, The Netherlands.
authors.forename: V M J
authors.initials: VM
authors.lastname: Verhoef
authors.affiliation: Department of Pathology, Radboud University Medical Center, Nijmegen, The Netherlands.
authors.forename: J
authors.initials: J
authors.lastname: Bulten
authors.affiliation: Department of Medical Microbiology, Radboud University Medical Center, Nijmegen, The Netherlands.
authors.forename: W J G
authors.initials: WJ
authors.lastname: Melchers
authors.affiliation: Department of Gynaecologic Oncology, Cancer Research Center, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands.
authors.forename: A G J
authors.initials: AG
authors.lastname: van der Zee
authors.affiliation: Department of Obstetrics and Gynaecology, Radboud University Medical Center, Nijmegen, The Netherlands.
authors.forename: R L M
authors.initials: RL
authors.lastname: Bekkers
authors.affiliation: Department of Gynaecologic Oncology, Cancer Research Center, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands.
authors.forename: G B A
authors.initials: GB
authors.lastname: Wisman
mesh_terms: D000328:Adult; D015815:Cell Adhesion Molecules / Q000235:genetics; D019175:DNA Methylation; D055088:Early Detection of Cancer / Q000379:methods; D005260:Female; D006801:Humans; D008840:Microfilament Proteins / Q000235:genetics; D008875:Middle Aged; D030361:Papillomavirus Infections / Q000821:virology*; D010349:Patient Compliance; D012307:Risk Factors; D012648:Self Care; D012680:Sensitivity and Specificity; D013048:Specimen Handling; D019098:Telomerase / Q000235:genetics; D014218:Triage / Q000379:methods*; D025521:Tumor Suppressor Proteins / Q000235:genetics; D002583:Uterine Cervical Neoplasms / Q000175:diagnosis* / Q000821:virology; D014626:Vaginal Smears / Q000379:methods*; D002578:Uterine Cervical Dysplasia / Q000175:diagnosis* / Q000821:virology
chemical_list: D015815:Cell Adhesion Molecules; C116887:EPB41L3 protein, human; C448000:JAM3 protein, human; D008840:Microfilament Proteins; C585115:RUBCNL protein, human; D025521:Tumor Suppressor Proteins; C509186:TERT protein, human; D019098:Telomerase