Resequencing microarray technology for genotyping human papillomavirus in cervical smears

• Published in: PloS one 2014-11, Vol.9 (11), p.e109301-e109301
• Main Authors: Berthet, Nicolas, Falguières, Michael, Filippone, Claudia, Bertolus, Chloé, Bole-Feysot, Christine, Brisse, Sylvain, Gessain, Antoine, Heard, Isabelle, Favre, Michel
• Format: Article
• Language: English
• Subjects: Alphapapillomavirus - genetics; Anogenital; Base Sequence; Biology and Life Sciences; Carcinoma; Cervix; Cluster Analysis; Deoxyribonucleic acid; Diagnostic systems; DNA; DNA microarrays; DNA Primers - genetics; DNA, Viral - genetics; Epidemiology; Female; Genomics; Genotype; Genotypes; Genotyping; Health aspects; Human health and pathology; Human papillomavirus; Humans; Identification; Infections; Influenza; Life Sciences; Medicine and Health Sciences; Methods; Molecular Sequence Data; Oligonucleotide Array Sequence Analysis - methods; Orthomyxoviridae; Papillomaviridae; Papillomavirus infections; Phylogeny; Public health; Real-Time Polymerase Chain Reaction; Research and Analysis Methods; Sequence Analysis, DNA; Sequences; Sexually transmitted diseases; STD; Technology; Vaccination; Vaginal Smears - methods; Viruses; Women's health
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0109301

There are more than 40 human papillomaviruses (HPVs) belonging to the alpha genus that cause sexually transmitted infections; these infections are among the most frequent and can lead to condylomas and anogenital intra-epithelial neoplasia. At least 18 of these viruses are causative agents of anogenital carcinomas. We evaluated the performance of a resequencing microarray for the detection and genotyping of alpha HPV of clinical significance using cloned HPV DNA. To reduce the number of HPV genotypes tiled on microarray, we used reconstructed ancestral sequences (RASs) as they are more closely related to the various genotypes than the current genotypes are among themselves. The performance of this approach was tested by genotyping with a set of 40 cervical smears already genotyped using the commercial PapilloCheck kit. The results of the two tests were concordant for 70% (28/40) of the samples and compatible for 30% (12/40). Our findings indicate that RASs were able to detect and identify one or several HPV in clinical samples. Associating RASs with homonym sequences improved the genotyping of HPV present in cases of multiple infection. In conclusion, we demonstrate the diagnostic potential of resequencing technology for genotyping of HPV, and illustrate its value both for epidemiological studies and for monitoring the distribution of HPV in the post-vaccination era.