Simultaneous amplification and identification of 25 human papillomavirus types with Templex technology
Simultaneous amplification and identification of 25 human papillomavirus types with Templex technology
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DOI：
10.1128/jcm.01762-06
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发表时间：
2006-11-01
影响因子：
9.4
通讯作者：
Tang, Yi-Wei
中科院分区：
文献类型：
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作者：
Han, Jian;Swan, David C.;Tang, Yi-Wei
大多数现有的人乳头瘤病毒（HPV）基因分型测定是基于使用共有或简并引物的多重PCR。我们开发了一种Templex HPV检测试剂盒，使用HPV特异性E6和E7基因的类型特异性引物，在单管反应中同时检测和鉴定25种常见的HPV基因型。对于HPV 16型（HPV-16）、-18型和-56型，Templex检测试剂盒的分析灵敏度分别为20、100和20拷贝/反应混合物。当多种HPV类型共存于一个反应中时，Templex检测试剂盒可提供每种类型的半定量信息。我们检测了109例临床宫颈标本，这些标本之前曾用Digene HC 2高危型HPV DNA检测进行过评估，结果一致率为95.4%。Templex检测提供了类型特异性结果，并在29.2%（48例中的14例）的高危HPV阳性样本中发现了多种类型。整个Templex程序，包括DNA提取，可以在5小时内完成，为HPV检测和分型提供了一种快速可靠的诊断工具，可以自动化。
The majority of existing human papillomavirus (HPV) genotyping assays are based on multiplex PCR using consensus or degenerate primers. We developed a Templex HPV assay that simultaneously detects and identifies 25 common HPV genotypes in a single-tube reaction using type-specific primers for the HPV-specific E6 and E7 genes. The analytical sensitivities of the Templex assay for HPV type 16 (HPV-16), -18, and -56 were 20, 100, and 20 copies per reaction mixture, respectively. The Templex assay provides semiquantitative information on each type when multiple HPV types coexist in one reaction. We tested 109 clinical cervical specimens previously evaluated with the Digene HC2 high-risk HPV DNA test and found 95.4% concordance between the assay results. The Templex assay provided type-specific results and found multiple types in 29.2% (14 of 48) of high-risk HPV-positive samples. The entire Templex procedure, including DNA extraction, can be completed within 5 hours, providing a rapid and reliable diagnostic tool for HPV detection and typing that is amenable to automation.