title: Clinical performance of RNA and DNA based HPV testing in a colposcopy setting: Influence of assay target, cut off and age.
abstract: BACKGROUND
  As HPV testing is used increasingly for cervical disease management, there is a demand to optimise the performance of HPV tests, particularly with respect to specificity.
  
  
  OBJECTIVE
  To compare the clinical performance of an HPV DNA and a RNA based test in women with cytological abnormalities. The influence of age and assay cut off on test performance was also assessed.
  
  
  METHODS
  A prospective comparison of the Hybrid Capture 2 test (HC2) and the Aptima HPV assay (AHPV) was performed within a colposcopy setting. Clinical sensitivity and specificity were determined for the detection of cervical intraepithelial neoplasia (CIN) grade 2 or worse.
  
  
  RESULTS
  Both assays were >90% sensitive for the detection of CIN2+. AHPV was slightly more specific than HC2 [49.9% (46.8-53.1) vs 45.9% (42.8, 49.1), p<0.0001]. Raising HC2 cut off to 2 RLU did not improve specificity. A cut-off of 10 RLU increased specificity by approximately 10% - although this led to a reduction in sensitivity of 6.3% which equated to 24 missed cases of CIN2+. Both assays were more specific in women over 30 years of age, compared to women under 30 (p<0.001).
  
  
  CONCLUSIONS
  Although AHPV was more specific than HC2 in the total cohort (p<0.001), we found this difference to be smaller than other studies. This could be attributed to different indications for colposcopic referral across different settings. This study also confirms the relatively poor specificity of commercial HPV assays in women under 30.
authors.affiliation: Scottish HPV Reference Laboratory, Royal Infirmary of Edinburgh, 51 Little France Crescent, Edinburgh EH16 4SA, United Kingdom. Electronic address: Kate.Cuschieri@luht.scot.nhs.uk.
authors.forename: Kate
authors.initials: K
authors.lastname: Cuschieri
authors.affiliation: HPV Research Group, University of Edinburgh MRC Centre for Reproductive Health, The Queen's Medical Research Institute, 47 Little France Crescent, Edinburgh EH16 4TJ, United Kingdom.
authors.forename: Heather
authors.initials: H
authors.lastname: Cubie
authors.affiliation: Epidemiology and Statistics Core, Wellcome Trust Clinical Research Facility, Edinburgh, United Kingdom.
authors.forename: Catriona
authors.initials: C
authors.lastname: Graham
authors.affiliation: Simpsons Centre for Reproductive Health, Royal Infirmary of Edinburgh, United Kingdom.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Rowan
authors.affiliation: Scottish HPV Reference Laboratory, Royal Infirmary of Edinburgh, 51 Little France Crescent, Edinburgh EH16 4SA, United Kingdom.
authors.forename: Alison
authors.initials: A
authors.lastname: Hardie
authors.affiliation: MRC Centre for Reproductive Health, University of Edinburgh, United Kingdom.
authors.forename: Andrew
authors.initials: A
authors.lastname: Horne
authors.affiliation: Epidemiology and Statistics Core, Wellcome Trust Clinical Research Facility, Edinburgh, United Kingdom.
authors.forename: Camille Busby
authors.initials: CB
authors.lastname: Earle
authors.affiliation: Manchester Medical Microbiology Partnership, Manchester Royal Infirmary, United Kingdom.
authors.forename: Andrew
authors.initials: A
authors.lastname: Bailey
authors.affiliation: Institute of Cancer Sciences, University of Manchester, St Mary's Hospital, Manchester, United Kingdom.
authors.forename: Emma J
authors.initials: EJ
authors.lastname: Crosbie
authors.affiliation: Institute of Cancer Sciences, University of Manchester, St Mary's Hospital, Manchester, United Kingdom.
authors.forename: Henry
authors.initials: H
authors.lastname: Kitchener
mesh_terms: D000328:Adult; D003127:Colposcopy / Q000379:methods*; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D055088:Early Detection of Cancer / Q000379:methods*; D005260:Female; D006801:Humans; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D027383:Papillomaviridae / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000150:complications / Q000175:diagnosis* / Q000821:virology; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D055815:Young Adult
chemical_list: D004279:DNA, Viral; D012367:RNA, Viral
keywords: AHPV; Aptima HPV test; CIN; Cervical screening; Clinical-performance; Diagnostics; HC2; HPV; Hybrid Capture 2 test; cervical intraepithelial neoplasia; human papillomavirus