title: Easy and fast detection and genotyping of high-risk human papillomavirus by dedicated DNA microarrays.
abstract: Persistent cervical high-risk human papillomavirus (HPV) infection is correlated with an increased risk of developing a high-grade cervical intraepithelial lesion. A two-step method was developed for detection and genotyping of high-risk HPV. DNA was firstly amplified by asymmetrical PCR in the presence of Cy3-labelled primers and dUTP. Labelled DNA was then genotyped using DNA microarray hybridization. The current study evaluated the technical efficacy of laboratory-designed HPV DNA microarrays for high-risk HPV genotyping on 57 malignant and non-malignant cervical smears. The approach was evaluated for a broad range of cytological samples: high-grade squamous intraepithelial lesions (HSIL), low-grade squamous intraepithelial lesions (LSIL) and atypical squamous cells of high-grade (ASC-H). High-risk HPV was also detected in six atypical squamous cells of undetermined significance (ASC-US) samples; among them only one cervical specimen was found uninfected, associated with no histological lesion. The HPV oligonucleotide DNA microarray genotyping detected 36 infections with a single high-risk HPV type and 5 multiple infections with several high-risk types. Taken together, these results demonstrate the sensitivity and specificity of the HPV DNA microarray approach. This approach could improve clinical management of patients with cervical cytological abnormalities.
authors.affiliation: Laboratoire de Virologie, Hôpital de l'Archet, BP 3079, 151 route de Saint-Antoine de Ginestière, 06200 Nice Cedex 3, France.
authors.forename: Valérie
authors.initials: V
authors.lastname: Albrecht
authors.forename: Anne
authors.initials: A
authors.lastname: Chevallier
authors.forename: Virginie
authors.initials: V
authors.lastname: Magnone
authors.forename: Pascal
authors.initials: P
authors.lastname: Barbry
authors.forename: Fanny
authors.initials: F
authors.lastname: Vandenbos
authors.forename: André
authors.initials: A
authors.lastname: Bongain
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Lefebvre
authors.forename: Valérie
authors.initials: V
authors.lastname: Giordanengo
mesh_terms: D001483:Base Sequence; D002232:Carbocyanines / Q000378:metabolism; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis* / Q000235:genetics; D003856:Deoxyuracil Nucleotides / Q000378:metabolism; D005838:Genotype; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D027383:Papillomaviridae / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000175:diagnosis / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D014626:Vaginal Smears
chemical_list: D002232:Carbocyanines; D017931:DNA Primers; D004279:DNA, Viral; D003856:Deoxyuracil Nucleotides; C089187:cyanine dye 3; C027078:deoxyuridine triphosphate