title: Comparison of oncogenic HPV type-specific viral DNA load and E6/E7 mRNA detection in cervical samples: results from a multicenter study.
abstract: High-risk human papillomavirus (HR-HPV) genotype viral load and E6/E7 mRNA detection are proposed as surrogate markers of malignant cervical lesion progression. Currently, the use of commercially available DNA-based or mRNA-based tests is under investigation. In this study, the viral DNA load and E6/E7 mRNA detection of the five most common HR-HPV types detected in cervical cancer worldwide were compared in 308 cervical samples by using in-house type-specific quantitative real-time PCR assays and PreTect HPV-Proofer test, respectively. Sensitivity and negative predictive values were higher for the HPV-DNA assays combined (95.0% and 96.0%, respectively) than the RNA assays (77.0% and 88.0%, respectively); conversely, the mRNA test showed a higher specificity and higher positive predictive value (81.7% and 66.9%, respectively) than the DNA test (58.6% and 52.5%, respectively) for detecting histology-confirmed high-grade cervical intraepithelial neoplasia. A significantly higher association between viral DNA load and severity of disease was observed for HPV 16 and 31 (γ = 0.62 and γ = 0.40, respectively) than for the other HPV types screened. A good degree of association between the two assays was found for detection of HPV 16 (k = 0.83), HPV 18 (k = 0.72), HPV 33 (k = 0.66), and HPV 45 (k = 0.60) but not for HPV 31 (k = 0.24). Sequence analysis in L1 and E6-LCR regions of HPV 31 genotypes showed a high level of intra-type variation. HR-HPV viral DNA load was significantly higher in E6/E7 mRNA positive than negative samples (P < 0.001), except for HPV 31. These findings suggest that transcriptional and replicative activities can coexist within the same sample.
authors.affiliation: Department of Health Sciences, University of Milano-Bicocca, Milan, Italy. francesco.broccolo@unimib.it
authors.forename: Francesco
authors.initials: F
authors.lastname: Broccolo
authors.forename: Lisa
authors.initials: L
authors.lastname: Fusetti
authors.forename: Sandra
authors.initials: S
authors.lastname: Rosini
authors.forename: Donatella
authors.initials: D
authors.lastname: Caraceni
authors.forename: Roberta
authors.initials: R
authors.lastname: Zappacosta
authors.forename: Lucia
authors.initials: L
authors.lastname: Ciccocioppo
authors.forename: Barbara
authors.initials: B
authors.lastname: Matteoli
authors.forename: Philippe
authors.initials: P
authors.lastname: Halfon
authors.forename: Mauro S
authors.initials: MS
authors.lastname: Malnati
authors.forename: Luca
authors.initials: L
authors.lastname: Ceccherini-Nelli
mesh_terms: D000328:Adult; D000368:Aged; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D005260:Female; D005838:Genotype; D006801:Humans; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D027383:Papillomaviridae / Q000235:genetics / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000150:complications / Q000175:diagnosis / Q000821:virology; D011237:Predictive Value of Tests; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012189:Retrospective Studies; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D002583:Uterine Cervical Neoplasms / Q000175:diagnosis* / Q000821:virology*; D055815:Young Adult
chemical_list: D004279:DNA, Viral; D012333:RNA, Messenger; D012367:RNA, Viral