title: Highly effective detection of human papillomavirus 16 and 18 DNA by a testing algorithm combining broad-spectrum and type-specific PCR.
abstract: The use of a single broad-spectrum human papillomavirus (HPV) DNA-based PCR test may fail to detect lower concentrations of HPV DNA due to competition between different genotypes in mixed infections. To improve HPV detection by PCR, broad-spectrum and type-specific (TS) PCRs were combined, with a focus on HPV-16 and HPV-18. Cervical and cervicovaginal cell samples were obtained from 1,113 healthy women (age range, 15 to 25 years) participating in an HPV-16/HPV-18 candidate vaccine efficacy trial. These samples were tested by a broad-spectrum SPF(10) PCR-DNA enzyme immunoassay, followed by a primer SPF(10)-based line probe assay (SPF(10) LiPA), and HPV-16- and HPV-18-TS PCRs. The results for the majority of the HPV-16/18 SPF(10) LiPA-positive samples were confirmed by TS-PCR (kappa values, 0.775 for HPV-16 and 0.785 for HPV-18). However, TS PCR revealed additional positive samples among those that contained other HPV genotypes due to competition. Conversely, SPF(10) LiPA identified HPV-16 or -18 in samples that remained negative by TS PCR as a result of sampling variation. Analysis of follow-up samples from more than 1,000 women confirmed that the combination of SPF(10)-LiPA with additional HPV-16- and HPV-18-TS PCR diminishes the rate of false-negative diagnosis. The combination of broad-spectrum and TS PCRs resulted in a novel testing algorithm. This combination of assays is more accurate than either method alone, and the novel algorithm offers a highly accurate and effective method for the analysis of HPV infections.
authors.affiliation: DDL Diagnostic Laboratory, Fonteijnenburghlaan 5, 2275 CX, Voorburg, The Netherlands. L.J.van.Doorn@ddl.nl
authors.forename: Leen-Jan
authors.initials: LJ
authors.lastname: van Doorn
authors.forename: Anco
authors.initials: A
authors.lastname: Molijn
authors.forename: Bernhard
authors.initials: B
authors.lastname: Kleter
authors.forename: Wim
authors.initials: W
authors.lastname: Quint
authors.forename: Brigitte
authors.initials: B
authors.lastname: Colau
mesh_terms: D000293:Adolescent; D000328:Adult; D000465:Algorithms; D002584:Cervix Uteri / Q000821:virology; D015344:DNA Probes, HPV; D004279:DNA, Viral / Q000032:analysis* / Q000302:isolation & purification; D005260:Female; D052162:Human papillomavirus 16 / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D052161:Human papillomavirus 18 / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D030361:Papillomavirus Infections / Q000175:diagnosis* / Q000517:prevention & control / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods*; D013045:Species Specificity; D016896:Treatment Outcome; D014765:Viral Vaccines / Q000627:therapeutic use
chemical_list: D015344:DNA Probes, HPV; D004279:DNA, Viral; D014765:Viral Vaccines