临床样本中15种人乳头瘤病毒基因型的不对称GP5+/6+聚合酶链反应及荧光偏振检测杂交
Asymmetric GP5+/6+ PCR and hybridization with fluorescence polarization assay of 15 human papillomavirus genotypes in clinical samples.
作者信息
Ju Zhang, Yane Gao, Ding Li, Guowu Yin, Yanhai Guo, Yonglan Liu
机构信息
Institute of Gene Diagnosis, State Key Laboratory of Cancer Biology, Fourth Military Medical University, 17 Changle West Road, Xian, Shaanxi 710032, China.
出版信息
J Clin Virol. 2009 Feb;44(2):106-10. doi: 10.1016/j.jcv.2008.12.003. Epub 2009 Jan 13.
BACKGROUND
The detection of a broad spectrum of HPV genotypes has not been used widely in cervical cancer screening due to technical difficulties and high costs.
OBJECTIVES
To develop an asymmetric GP5+/6+ PCR assay and hybridization with a fluorescence polarization (FP) assay of 15 HPV genotypes.
STUDY DESIGN
HPV genes in controls and samples were amplified by an asymmetric GP5+/6+ PCR. Fifteen HPV genotypic probes labeled with fluorophores hybridized with target PCR product to identify the presence of specific HPV genotypes. The HPV genotypes in samples were verified by sequence assay.
RESULTS
The genotypes determined with the hybridization and FP assay were confirmed by sequence analysis when a monotypic infection was evaluated.
CONCLUSION
A simple, economical and specific HPV genotyping assay has been developed that will be adequate for cervical cancer screening programs.
背景
由于技术困难和成本高昂，广泛检测多种HPV基因型尚未在宫颈癌筛查中广泛应用。
目的
开发一种不对称GP5+/6+ PCR检测方法，并与15种HPV基因型的荧光偏振（FP）检测方法进行杂交。
研究设计
通过不对称GP5+/6+ PCR扩增对照和样本中的HPV基因。15种用荧光团标记的HPV基因型探针与目标PCR产物杂交，以鉴定特定HPV基因型的存在。通过序列分析验证样本中的HPV基因型。
结果
当评估单一型感染时，通过杂交和FP检测确定的基因型经序列分析得到证实。
结论
已开发出一种简单、经济且特异的HPV基因分型检测方法，适用于宫颈癌筛查项目。
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