Novel multiplexed genotyping of human papillomavirus using a VeraCode-allele-specific primer extension method.
Novel multiplexed genotyping of human papillomavirus using a VeraCode-allele-specific primer extension method.
复制标题
使用 VeraCode 等位基因特异性引物延伸方法对人乳头瘤病毒进行新型多重基因分型。
DOI：
10.1111/j.1348-0421.2011.00406.x
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
and Kukimoto I.
中科院分区：
文献类型：
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作者：
Kitamura-Muramatsu Y.;Kusumoto-Matsuo R.;and Kukimoto I.
将VeraCode等位基因特异性引物延伸（ASPE）方法应用于人乳头瘤病毒（HPV）-DNA的检测和基因分型。将含有HPV型特异性L1序列的寡核苷酸引物与通过PGMY-PCR扩增的HPV-DNA退火，然后通过ASPE用生物素化核苷酸标记DNA。通过VeraCode珠通过杂交捕获标记的DNA，用链霉亲和素缀合的荧光团染色，并通过Illumina BeadXpress®读数器检测。通过使用该系统，16种临床上重要的HPV类型（HPV 6、11、16、18、31、33、35、39、45、51、52、56、58、59、66和68）以多重格式被正确地基因分型。临床DNA样本的VeraCode‐ASPE基因分型结果与经验证的PGMY‐反向印迹杂交检测结果完全一致，为HPV流行病学调查所需的高通量基因分型提供了新平台。
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.