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Novel multiplexed genotyping of human papillomavirus using a VeraCode‐allele‐specific primer extension method
Author(s) -
KitamuraMuramatsu Yuri,
KusumotoMatsuo Rika,
Kondo Kazunari,
Mori Seiichiro,
Saito Susumu,
Tsukahara Yusuke,
Kukimoto Iwao
Publication year - 2012
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.2011.00406.x
Subject(s) - genotyping , biology , primer extension , multiplex , biotinylation , microbiology and biotechnology , oligonucleotide , multiplex polymerase chain reaction , genotype , primer (cosmetics) , dna , polymerase chain reaction , genetics , gene , base sequence , chemistry , organic chemistry
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.