title: Type-specific human papillomavirus E6/E7 mRNA detection by real-time PCR improves identification of cervical neoplasia.
abstract: DNA-based human papillomavirus (HPV) assays show high sensitivity but poor specificity in detecting high-grade cervical lesions. Assays detecting mRNA of the oncoproteins E6 and E7 show higher specificity but lack either detection of all high-risk HPV genotypes or the capacity to specify the detected genotypes. Therefore, a real-time PCR assay detecting type-specific E6/E7 mRNA was developed and the clinical performance evaluated. A total of 210 cervical LBC (liquid-based cytology) samples from 204 women were analyzed for HPV DNA and mRNA with the in-house real-time PCR as well as PreTect HPV-Proofer. The sensitivity of real-time PCR mRNA detection to identify histologically confirmed CIN2+ (cervical intraepithelial neoplasia, grade 2 or higher) was 0.91, compared to 0.95 for DNA analysis. The specificity was 0.68 compared to 0.38, and the positive predictive value (PPV) was higher for mRNA (0.67 versus 0.52) without any loss in negative predictive value (NPV). The sensitivity of the real-time PCR mRNA test was somewhat higher than that for PreTect HPV-Proofer (0.83 versus 0.75) in analyses for the same genotypes. The specificities were similar (0.76 versus 0.77). In analyses for mRNA of the eight most common genotypes in cervical cancer (HPV16, -18, -31, -33, -35, -45, -52, and -58), the sensitivity of detection of CIN2+ lesions was 0.87 and the specificity 0.74, with a PPV of 0.70. In conclusion, real-time PCR for detection of HPV E6/E7 mRNA transcripts can be a sensitive and specific tool in screening and investigation of cervical neoplasia. The composition of HPV types in mRNA testing needs to be further investigated to optimize sensitivity and specificity.
authors.affiliation: Department of Clinical Virology, University of Gothenburg, Gothenburg, Sweden. elin.andersson@microbio.gu.se
authors.forename: Elin
authors.initials: E
authors.lastname: Andersson
authors.forename: Cecilia
authors.initials: C
authors.lastname: Kärrberg
authors.forename: Thomas
authors.initials: T
authors.lastname: Rådberg
authors.forename: Lennart
authors.initials: L
authors.lastname: Blomqvist
authors.forename: Britt-Marie
authors.initials: BM
authors.lastname: Zetterqvist
authors.forename: Walter
authors.initials: W
authors.lastname: Ryd
authors.forename: Magnus
authors.initials: M
authors.lastname: Lindh
authors.forename: Peter
authors.initials: P
authors.lastname: Horal
mesh_terms: D000328:Adult; D000368:Aged; D055088:Early Detection of Cancer / Q000379:methods; D005260:Female; D006801:Humans; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D027383:Papillomaviridae / Q000235:genetics / Q000302:isolation & purification*; D011247:Pregnancy; D012333:RNA, Messenger / Q000235:genetics* / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D002583:Uterine Cervical Neoplasms / Q000175:diagnosis* / Q000821:virology
chemical_list: D009856:Oncogene Proteins, Viral; D012333:RNA, Messenger