title: Novel multiplexed genotyping of human papillomavirus using a VeraCode-allele-specific primer extension method.
abstract: A VeraCode-allele-specific primer extension (ASPE) method was applied to the detection and genotyping of human papillomavirus (HPV)-DNA. Oligonucleotide primers containing HPV-type-specific L1 sequences were annealed to HPV-DNA amplified by PGMY-PCR, followed by ASPE to label the DNA with biotinylated nucleotides. The labeled DNA was captured by VeraCode beads through hybridization, stained with a streptavidin-conjugated fluorophore, and detected by an Illumina BeadXpress® reader. By using this system, 16 clinically important HPV types (HPV6, 11, 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66 and 68) were correctly genotyped in a multiplex format. The VeraCode-ASPE genotyping of clinical DNA samples yielded identical results with those obtained by validated PGMY-reverse blot hybridization assay, providing a new platform for high-throughput genotyping required for HPV epidemiological surveys.
authors.affiliation: Riken Genesis Co., Ltd, Riken Yokohama Institute, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan.
authors.forename: Yuri
authors.initials: Y
authors.lastname: Kitamura-Muramatsu
authors.forename: Rika
authors.initials: R
authors.lastname: Kusumoto-Matsuo
authors.forename: Kazunari
authors.initials: K
authors.lastname: Kondo
authors.forename: Seiichiro
authors.initials: S
authors.lastname: Mori
authors.forename: Susumu
authors.initials: S
authors.lastname: Saito
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Tsukahara
authors.forename: Iwao
authors.initials: I
authors.lastname: Kukimoto
mesh_terms: D000483:Alleles; D052685:Alphapapillomavirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers / Q000235:genetics; D005260:Female; D005838:Genotype; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D030361:Papillomavirus Infections / Q000175:diagnosis / Q000821:virology*
chemical_list: D017931:DNA Primers