title: Development of a bead-based multiplex genotyping method for diagnostic characterization of HPV infection.
abstract: The accurate genotyping of human papillomavirus (HPV) is clinically important because the oncogenic potential of HPV is dependent on specific genotypes. Here, we described the development of a bead-based multiplex HPV genotyping (MPG) method which is able to detect 20 types of HPV (15 high-risk HPV types 16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68 and 5 low-risk HPV types 6, 11, 40, 55, 70) and evaluated its accuracy with sequencing. A total of 890 clinical samples were studied. Among these samples, 484 were HPV positive and 406 were HPV negative by consensus primer (PGMY09/11) directed PCR. The genotyping of 484 HPV positive samples was carried out by the bead-based MPG method. The accuracy was 93.5% (95% CI, 91.0-96.0), 80.1% (95% CI, 72.3-87.9) for single and multiple infections, respectively, while a complete type mismatch was observed only in one sample. The MPG method indiscriminately detected dysplasia of several cytological grades including 71.8% (95% CI, 61.5-82.3) of ASCUS (atypical squamous cells of undetermined significance) and more specific for high grade lesions. For women with HSIL (high grade squamous intraepithelial lesion) and SCC diagnosis, 32 women showed a PPV (positive predictive value) of 77.3% (95% CI, 64.8-89.8). Among women >40 years of age, 22 women with histological cervical cancer lesions showed a PPV of 88% (95% CI, 75.3-100). Of the highest risk HPV types including HPV-16, 18 and 31 positive women of the same age groups, 34 women with histological cervical cancer lesions showed a PPV of 77.3% (95% CI, 65.0-89.6). Taken together, the bead-based MPG method could successfully detect high-grade lesions and high-risk HPV types with a high degree of accuracy in clinical samples.
authors.affiliation: Department of Anesthesiology, College of Medicine, The Catholic University of Korea, Seocho-ku, Seoul, Korea.
authors.forename: Mee Young
authors.initials: MY
authors.lastname: Chung
authors.forename: Yong-Wan
authors.initials: YW
authors.lastname: Kim
authors.forename: Su Mi
authors.initials: SM
authors.lastname: Bae
authors.forename: Eun Hye
authors.initials: EH
authors.lastname: Kwon
authors.forename: Pankaj Kumar
authors.initials: PK
authors.lastname: Chaturvedi
authors.forename: Gantumur
authors.initials: G
authors.lastname: Battogtokh
authors.forename: Woong Shick
authors.initials: WS
authors.lastname: Ahn
mesh_terms: D000328:Adult; D000368:Aged; D005260:Female; D005814:Genes, Viral; D005821:Genetic Techniques; D005838:Genotype; D006801:Humans; D008875:Middle Aged; D008957:Models, Genetic; D009693:Nucleic Acid Hybridization; D027383:Papillomaviridae / Q000235:genetics*; D030361:Papillomavirus Infections / Q000175:diagnosis* / Q000235:genetics*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013045:Species Specificity; D002583:Uterine Cervical Neoplasms / Q000175:diagnosis / Q000235:genetics