title: The diagnostic accuracy of two methods for E6&7 mRNA detection in women with minor cytological abnormalities.
abstract: OBJECTIVE
  To examine the diagnostic accuracy of nucleic acid sequence based amplification (NASBA) and flow cytometry for E6&7 human papillomavirus (HPV) mRNA detection in the triage of minor cytological abnormalities.
  
  
  METHODS
  Prospective diagnostic accuracy study.
  
  
  METHODS
  Gynecology outpatient clinics of a university hospital.
  
  
  METHODS
  472 women with low-grade squamous intraepithelial lesion (LSIL) or atypical squamous cells of undetermined significance (ASCUS).
  
  
  METHODS
  Residual material of the liquid-based smears was tested by NASBA and by flow cytometry for E6&E7 mRNA expression. Histological diagnosis was used as reference standard.
  
  
  METHODS
  Accuracy indices of the two techniques and of type 16-specific NASBA for the detection of cervical intraepithelial neoplasia (CIN) 2+ and CIN3+, accuracy indices at age >35 years, correlation between NASBA and flow, comparison between integrated and episomal high-risk HPV infection for risk of CIN2+.
  
  
  RESULTS
  Both tests showed increased positivity rates with increasing severity of the lesion (p < 0.05, chi-squared test for trend). There was a positive correlation between NASBA and flow results (phi coefficient = 0.325). NASBA-positive cases were more likely to have CIN2+ than were NASBA-negative/DNA-positive for types 16, 18, 31, 33, 45 (25/73 vs. 4/52, p= 0.0004; Fisher's exact test). In the LSIL group the NASBA accuracy indices for CIN3+ were: sensitivity 75%, specificity 78.7% and positivity rate 20.8%, and for flow 77.8%, 64.5% and 35.9%, respectively.
  
  
  CONCLUSIONS
  NASBA has favorable specificity and positivity rates for triaging LSIL prior to colposcopy. A relatively low sensitivity warrants cytological surveillance of the NASBA-negative LSILs. Flow cytometry does not perform as well overall.
authors.affiliation: 3rd Department of Obstetrics and Gynecology, Attikon University Hospital, Athens, Greece. georgekoliopoulos@yahoo.com
authors.forename: George
authors.initials: G
authors.lastname: Koliopoulos
authors.forename: Charalambos
authors.initials: C
authors.lastname: Chrelias
authors.forename: Asimakis
authors.initials: A
authors.lastname: Pappas
authors.forename: Sofia
authors.initials: S
authors.lastname: Makridima
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Kountouris
authors.forename: Maria
authors.initials: M
authors.lastname: Alepaki
authors.forename: Aris
authors.initials: A
authors.lastname: Spathis
authors.forename: Vicky
authors.initials: V
authors.lastname: Stathopoulou
authors.forename: Ioannis
authors.initials: I
authors.lastname: Panayiotides
authors.forename: Pericles
authors.initials: P
authors.lastname: Panagopoulos
authors.forename: Petros
authors.initials: P
authors.lastname: Karakitsos
authors.forename: Dimitrios
authors.initials: D
authors.lastname: Kassanos
mesh_terms: D000328:Adult; D003937:Diagnosis, Differential; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D050725:Papillomavirus E7 Proteins / Q000235:genetics*; D030361:Papillomavirus Infections / Q000175:diagnosis*; D011446:Prospective Studies; D012333:RNA, Messenger / Q000032:analysis*; D012097:Repressor Proteins / Q000235:genetics*; D021161:Self-Sustained Sequence Replication / Q000379:methods*; D012680:Sensitivity and Specificity; D014218:Triage; D002578:Uterine Cervical Dysplasia / Q000821:virology; D002583:Uterine Cervical Neoplasms / Q000821:virology*
chemical_list: C068651:E6 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D012333:RNA, Messenger; D012097:Repressor Proteins