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 b...

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Bibliographic Details
Published in:Microbiology and immunology 2012-02, Vol.56 (2), p.128-133
Main Authors: Kitamura-Muramatsu, Yuri, Kusumoto-Matsuo, Rika, Kondo, Kazunari, Mori, Seiichiro, Saito, Susumu, Tsukahara, Yusuke, Kukimoto, Iwao
Format: Article
Language:English
Subjects:
allele-specific primer extension
Alleles
Alphapapillomavirus - classification
Alphapapillomavirus - genetics
Alphapapillomavirus - isolation & purification
DNA Primers - genetics
Female
Genotype
genotyping
human papillomavirus
Humans
multiplex assay
Multiplex Polymerase Chain Reaction - instrumentation
Multiplex Polymerase Chain Reaction - methods
Papillomavirus Infections - diagnosis
Papillomavirus Infections - virology
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Summary: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.
ISSN:0385-5600
1348-0421
DOI:10.1111/j.1348-0421.2011.00406.x