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Novel multiplexed genotyping of human papillomavirus using a VeraCode-allele-specific primer extension method.

Authors
  • Kitamura-Muramatsu, Yuri
  • Kusumoto-Matsuo, Rika
  • Kondo, Kazunari
  • Mori, Seiichiro
  • Saito, Susumu
  • Tsukahara, Yusuke
  • Kukimoto, Iwao
Type
Published Article
Journal
Microbiology and immunology
Publication Date
Feb 01, 2012
Volume
56
Issue
2
Pages
128–133
Identifiers
DOI: 10.1111/j.1348-0421.2011.00406.x
PMID: 22146070
Source
Medline
License
Unknown

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.

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