Han Jian, Swan David C, Smith Sharon J, Lum Shanjuan H, Sefers Susan E, Unger Elizabeth R, Tang Yi-Wei
Genaco Biomedical Products, Inc., Huntsville, Alabama 35805, USA.
J Clin Microbiol. 2006 Nov;44(11):4157-62. doi: 10.1128/JCM.01762-06. Epub 2006 Sep 27.
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.
大多数现有的人乳头瘤病毒(HPV)基因分型检测方法是基于使用共有引物或简并引物的多重PCR。我们开发了一种Templex HPV检测方法,该方法使用针对HPV特异性E6和E7基因的型特异性引物,在单管反应中同时检测和鉴定25种常见的HPV基因型。Templex检测方法对HPV 16型(HPV-16)、-18型和-56型的分析灵敏度分别为每个反应混合物20、100和20个拷贝。当一种反应中同时存在多种HPV类型时,Templex检测方法可提供每种类型的半定量信息。我们对109份先前用Digene HC2高危HPV DNA检测评估过的临床宫颈标本进行了检测,发现检测结果之间的一致性为95.4%。Templex检测方法提供了型特异性结果,在29.2%(48份中的14份)的高危HPV阳性样本中发现了多种类型。整个Templex操作流程,包括DNA提取,可在5小时内完成,为HPV检测和分型提供了一种快速可靠且适合自动化的诊断工具。