title: Use of the HPV MLPA assay in cervical cytology for the prediction of high grade lesions.
abstract: Current screening methods for uterine cervical cancer such as Papanicolaou smears and/or high risk human Papillomavirus (HR-HPV) detection have a high negative predictive value but a low positive predictive value for the presence of high grade cervical lesions. Therefore, new parameters are needed to reduce the rate of unnecessary referrals for colposcopy. The predictive value of the HPV multiplex ligation-dependent probe amplification (MLPA) assay, which can assess simultaneously HPV16/18 viral load and viral integration, was evaluated. The assay was applied to 170 cervical cytological samples, and the results were correlated with the matching histological follow-up. The GP5+/6+ assay and qPCR were used as a control for HR-HPV typing. The MLPA assay classified a higher percentage of cases as high-risk (high-viral load and/or viral integration) with higher grades of dysplasia. There was a high correlation between the HPV MLPA assay and qPCR for viral load and HPV genotyping, and between the MLPA assay and the GP5+/6+ assay for HPV genotyping. The sensitivity and specificity of the HPV MLPA assay for the detection of high-grade lesions were 44% and 93%, respectively. This study demonstrates that the HPV MLPA assay can reliably detect HPV 16/18, viral load, and viral integration in cytological samples. Also, high-risk classification correlated well with the presence of high-grade dysplasia. However, for the implementation of the MLPA assay into clinical practice, additional HR-HPV types need to be included to increase the sensitivity of the assay, and thereby increase its negative predictive value.
authors.affiliation: Department of Molecular Cell Biology, GROW-School for Oncology and Developmental Biology, Maastricht University Medical Center, Maastricht, The Netherlands.
authors.forename: Rogier J N T M
authors.initials: RJ
authors.lastname: Litjens
authors.forename: Wendy
authors.initials: W
authors.lastname: Theelen
authors.forename: Yvonne
authors.initials: Y
authors.lastname: van de Pas
authors.forename: Jessica
authors.initials: J
authors.lastname: Ossel
authors.forename: Martin
authors.initials: M
authors.lastname: Reijans
authors.forename: Guus
authors.initials: G
authors.lastname: Simons
authors.forename: Ernst-Jan M
authors.initials: EJ
authors.lastname: Speel
authors.forename: Brigitte F M
authors.initials: BF
authors.lastname: Slangen
authors.forename: Frans C S
authors.initials: FC
authors.lastname: Ramaekers
authors.forename: Roy F P M
authors.initials: RF
authors.lastname: Kruitwagen
authors.forename: Anton H N
authors.initials: AH
authors.lastname: Hopman
mesh_terms: D000328:Adult; D003584:Cytological Techniques / Q000379:methods*; D005260:Female; D006801:Humans; D008403:Mass Screening / Q000379:methods; D008875:Middle Aged; D027383:Papillomaviridae / Q000302:isolation & purification* / Q000472:pathogenicity / Q000502:physiology; D030361:Papillomavirus Infections / Q000150:complications* / Q000175:diagnosis*; D057089:Pathology, Molecular / Q000379:methods*; D012680:Sensitivity and Specificity; D002583:Uterine Cervical Neoplasms / Q000175:diagnosis* / Q000821:virology*; D019562:Viral Load; D016662:Virus Integration; D055815:Young Adult