Faculty of Medicine, University of Banja Luka, Banja Luka, Bosnia and Herzegovina.
Institute of Microbiology and Immunology, Faculty of Medicine, University of Belgrade, Belgrade, Republic of Serbia.
Eur J Oral Sci. 2020 Dec;128(6):501-507. doi: 10.1111/eos.12746. Epub 2020 Nov 17.
The potential problems of DNA extraction from formalin-fixed paraffin-embedded (FFPE) tissue samples and amplification efficiency of Human papilloma virus (HPV) may occur in the molecular studies of head and neck squamous carcinoma (HNSCC). The aim of this study was to compare HPV detection rate in FFPE tissues of oral, oropharyngeal, hypopharyngeal, and laryngeal cancers using two silica-based extraction kits and three amplification methods. A total of 50 FFPE specimens from HNSCC tissues were analyzed. The quality and quantity of the extracted DNA were tested by spectrophotometry. HPV DNA was detected using a single polymerase chain reaction (PCR), a nested PCR, and a Real-time PCR kit. Statistically significantly higher DNA quality and quantity was observed using the QIAamp DNA FFPE Tissue Kit than when using the QIAamp DNA Mini Kit. There was not HPV amplification in any of the 50 FFPE samples using the single PCR and Real-time PCR kits, whereas HPV DNA was detected in 22% of samples using nested PCR. Comparing results of the three different methods showed that HPV DNA was detected only with nested PCR. The results presented imply that nested PCR is the most appropriate method for the detection of HPV DNA in FFPE samples, along with adequate DNA extraction methods.
从福尔马林固定石蜡包埋（FFPE）组织样本中提取 DNA 以及人乳头瘤病毒（HPV）扩增效率可能会出现问题，这可能会影响头颈部鳞状细胞癌（HNSCC）的分子研究。本研究的目的是比较两种基于硅的提取试剂盒和三种扩增方法在口腔、口咽、下咽和喉癌 FFPE 组织中 HPV 检测率。共分析了 50 例 HNSCC 组织的 FFPE 标本。通过分光光度法测试提取 DNA 的质量和数量。使用单重聚合酶链反应（PCR）、嵌套 PCR 和实时 PCR 试剂盒检测 HPV DNA。与 QIAamp DNA Mini 试剂盒相比，使用 QIAamp DNA FFPE Tissue Kit 观察到 DNA 质量和数量有显著提高。使用单重 PCR 和实时 PCR 试剂盒在任何 50 个 FFPE 样本中均未扩增 HPV，而嵌套 PCR 检测到 22%的样本中存在 HPV DNA。比较三种不同方法的结果表明，只有嵌套 PCR 可检测 HPV DNA。结果表明，嵌套 PCR 是检测 FFPE 样本中 HPV DNA 的最适宜方法，同时需要适当的 DNA 提取方法。
