Easy and fast detection and genotyping of high-risk human papillomavirus by dedicated DNA microarrays.
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.
Albrecht
Valérie
V
Laboratoire de Virologie, Hôpital de l'Archet, BP 3079, 151 route de Saint-Antoine de Ginestière, 06200 Nice Cedex 3, France.
Chevallier
Anne
A
Magnone
Virginie
V
Barbry
Pascal
P
Vandenbos
Fanny
F
Bongain
André
A
Lefebvre
Jean-Claude
JC
Giordanengo
Valérie
V
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
D002232:Carbocyanines; D017931:DNA Primers; D004279:DNA, Viral; D003856:Deoxyuracil Nucleotides; C089187:cyanine dye 3; C027078:deoxyuridine triphosphate