title: Short fragment polymerase chain reaction reverse hybridization line probe assay to detect and genotype a broad spectrum of human papillomavirus types. Clinical evaluation and follow-up.
abstract: The purpose of this study was to detect and genotype 16 different human papilloma virus (HPV) types simultaneously using a short fragment polymerase chain reaction (SPF) hybridization line probe assay (LiPA). 152 women who were referred to the gynecologist because of abnormal cervical smear underwent colposcopic examination and repeat cervical smear. In addition, the cervical scrapes were analyzed for the presence of HPV by a novel general HPV polymerase chain reaction assay followed by a single reaction genotyping assay allowing for a simultaneous detection and identification of 16 different HPV types. HPV DNA was detected in 38% of normal follow-up cervical scrapes, 51% of scrapes with atypical squamous cells of undetermined significance, 78% of scrapes with mild dysplasia (low grade squamous intraepithelial lesions), 86% of scrapes with moderate dysplasia (high grade squamous intraepithelial lesions), and in 88% of scrapes with severe dysplasia and carcinoma in situ. One case of invasive squamous cell carcinoma was positive for HPV 16. Overall, a single HPV type was detected in 56% of HPV positive scrapes, with HPV 16 being the most common and accounting for 45% of all single infections. Forty-four percent of the positive scrapes contained multiple HPV types, of which double infections prevailed. Follow-up results proved the reproducibility and reliability of SPF HPV LiPA. In conclusion, we have used and evaluated the SPF-HPV-LiPA system for the detection and genotyping of HPV infections. The combined detection-typing method proved to be sensitive, specific, simple, and fast, making mass screening of cervical scrapes accessible for routine practice and facilitating individual patient management.
authors.affiliation: Department of Medical Microbiology, University of Nijmegen, Nijmegen Delft, The Netherlands. w.melchers@mmb.azn.nl
authors.forename: W J
authors.initials: WJ
authors.lastname: Melchers
authors.forename: J M
authors.initials: JM
authors.lastname: Bakkers
authors.forename: J
authors.initials: J
authors.lastname: Wang
authors.forename: P C
authors.initials: PC
authors.lastname: de Wilde
authors.forename: H
authors.initials: H
authors.lastname: Boonstra
authors.forename: W G
authors.initials: WG
authors.lastname: Quint
authors.forename: A G
authors.initials: AG
authors.lastname: Hanselaar
mesh_terms: D005260:Female; D016679:Genome, Viral; D006801:Humans; D008828:Microbiological Techniques; D009693:Nucleic Acid Hybridization / Q000379:methods; D027383:Papillomaviridae / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods