title: Evaluation of combined general primer-mediated PCR sequencing and type-specific PCR strategies for determination of human papillomavirus genotypes in cervical cell specimens.
abstract: A strategy combining human papillomavirus general primer (mainly the PGMY primers)-directed PCR sequencing and type-specific PCR is presented. DNA samples were first tested in general primer-mediated PCR. The amplified fragments of positive samples after ethidium bromide-stained DNA gel analysis were further sequenced, and corresponding DNA samples were further analyzed by PCR using type-specific primers for human papillomavirus (HPV) types 16, 18, 31, and 52. The comparison of the results of 157 samples analyzed by this strategy in parallel with the Hybrid Capture 2 tests and with the HPV INNO-LiPA (Innogenetics line probe assay) shows that this method is suitable for HPV detection and genotyping in cervical cell samples. Although the PCR sequencing method is as sensitive as the HPV INNO-LiPA for HPV detection, our method allows the identification of a broader range of HPV types. In contrast, the HPV INNO-LiPA was less time-consuming and better identified coinfections.
authors.affiliation: Laboratory of Molecular Virology, ISP/Institut Pasteur, rue Engeland 642, 1180 Brussels, Belgium. vfontaine@pasteur.be
authors.forename: Véronique
authors.initials: V
authors.lastname: Fontaine
authors.forename: Corinne
authors.initials: C
authors.lastname: Mascaux
authors.forename: Christine
authors.initials: C
authors.lastname: Weyn
authors.forename: Aurore
authors.initials: A
authors.lastname: Bernis
authors.forename: Nathalie
authors.initials: N
authors.lastname: Celio
authors.forename: Philippe
authors.initials: P
authors.lastname: Lefèvre
authors.forename: Leonard
authors.initials: L
authors.lastname: Kaufman
authors.forename: Christian
authors.initials: C
authors.lastname: Garbar
mesh_terms: D002584:Cervix Uteri / Q000166:cytology* / Q000821:virology*; D017931:DNA Primers; D005260:Female; D005838:Genotype; D052162:Human papillomavirus 16 / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D052161:Human papillomavirus 18 / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D027383:Papillomaviridae / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D013045:Species Specificity; D013048:Specimen Handling / Q000379:methods; D002583:Uterine Cervical Neoplasms / Q000821:virology
chemical_list: D017931:DNA Primers