title: A general primer GP5+/GP6(+)-mediated PCR-enzyme immunoassay method for rapid detection of 14 high-risk and 6 low-risk human papillomavirus genotypes in cervical scrapings.
abstract: Two cocktails of digoxigenin-labeled human papillomavirus (HPV) type-specific oligonucleotide probes and an enzyme immunoassay (EIA) were used as a basis to developed a group-specific detection method for 14 high-risk (types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68) and 6 low-risk (types 6, 11, 40, 42, 43, and 44) HPVs, following a general primer GP5+/bioGP6(+)-mediated PCR. The sensitivity of this high-risk/low-risk (HR/LR) HPV PCR-EIA ranged from 10 to 200 HPV copies, depending on the HPV type. Comparison of HR/LR HPV PCR-EIA with radioactive Southern blot hybridization using a general probe on the same PCR products derived from 417 cytomorphologically abnormal cervical scrapings resulted in an overall agreement of 96% between the two methods. Complete concordance between group-specific HR/LR detection and individual typing results for both single and multiple infections indicate the strong specificity of this HR/LR HPV PCR-EIA. Multiple infections could be predicted by comparing PCR-EIA optical density values of the cocktail probes with one of the individual oligonucleotide probes. This novel HR/LR PCR-EIA allows accurate and rapid identification of high-risk and low-risk HPV types in cervical scrapings and will facilitate HPV detection in HPV mass-screening programs.
authors.affiliation: Department of Pathology, University Hospital Vrije Universiteit, Amsterdam, The Netherlands.
authors.forename: M V
authors.initials: MV
authors.lastname: Jacobs
authors.forename: P J
authors.initials: PJ
authors.lastname: Snijders
authors.forename: A J
authors.initials: AJ
authors.lastname: van den Brule
authors.forename: T J
authors.initials: TJ
authors.lastname: Helmerhorst
authors.forename: C J
authors.initials: CJ
authors.lastname: Meijer
authors.forename: J M
authors.initials: JM
authors.lastname: Walboomers
mesh_terms: D001483:Base Sequence; D002584:Cervix Uteri / Q000821:virology*; D017931:DNA Primers / Q000235:genetics; D005260:Female; D005838:Genotype; D006801:Humans; D007124:Immunoenzyme Techniques / Q000706:statistics & numerical data; D008403:Mass Screening; D027383:Papillomaviridae / Q000235:genetics* / Q000302:isolation & purification* / Q000472:pathogenicity; D030361:Papillomavirus Infections / Q000150:complications / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D012307:Risk Factors; D012680:Sensitivity and Specificity; D014412:Tumor Virus Infections / Q000150:complications / Q000821:virology; D002577:Uterine Cervical Diseases / Q000150:complications / Q000821:virology; D002583:Uterine Cervical Neoplasms / Q000209:etiology / Q000517:prevention & control / Q000821:virology; D014773:Virology / Q000379:methods / Q000706:statistics & numerical data
chemical_list: D017931:DNA Primers