评估和验证 HPV 实时 PCR 检测方法在口腔刷和 FFPE 样本中 HPV DNA 的检测。
Evaluation and validation of HPV real-time PCR assay for the detection of HPV DNA in oral cytobrush and FFPE samples.
机构信息
Université de Lorraine, Nancy, France.
CNRS, UMR, 7039 CRAN, Nancy, France.
出版信息
Sci Rep. 2018 Jul 27;8(1):11313. doi: 10.1038/s41598-018-29790-z.
Specific HPV genotypes have been recognized as risk factors inducing head and neck cancers (HNC). The aim of this study was to validate a real-time PCR assay to detect accurately High Risk HPV DNA in Formalin Fixed Paraffin Embedded (FFPE) and oral cytobrush samples and compare the results with conventional PCR. Repeatability, reproducibility and limit of detection of Cobas assay were estimated for oral cytobrush and FFPE samples of patients with HNC. 53 samples of patients with a HNC were then used for assay comparison with conventional PCR. Finally, 26 samples of patients with anogenital neoplasia cancer were analyzed as control and assays comparison. Among the 53 samples of patients with HNC, 12 (26.7%) were HPV positive, 33 (73.3%) were HPV negative and 8 (15.1%) were non contributive with the Cobas assay. Among the 26 samples of patients with anogenital neoplasia, 15 (57.7%) were HPV positive and 11 were HPV negative (42.3%). One sample was found with an HPV 16 and HPV 18 co-infection. Only 3 samples were found with discrepant results. Cobas assay was found suitable for routine HPV detection with a very good repeatability and reproducibility for all HPV genotypes (CV < 0.6% and <0.4% respectively). Sensitivity and specificity for Cobas assay were 91.7% [61.5%;99.8%] and 96.9% [83.8%;99.9%] respectively. Ten nanograms of DNA were sufficient for the detection of HPV 16, HPV 18 and HPV in FFPE and oral cytobrush samples. Cobas assay was found comparable to conventional PCR and can detect accurately and rapidly HPV DNA in FFPE and oral cytobrush samples for the management of HNC and other types of HPV-associated neoplasia.
特定的 HPV 基因型已被认为是导致头颈部癌症（HNC）的危险因素。本研究旨在验证一种实时 PCR 检测方法，以准确检测福尔马林固定石蜡包埋（FFPE）和口腔刷样本中的高危 HPV DNA，并将结果与传统 PCR 进行比较。对 HNC 患者的口腔刷和 FFPE 样本进行了 Cobas 检测的重复性、重现性和检测限的评估。然后，将 53 例 HNC 患者的样本与传统 PCR 进行了比较。最后，分析了 26 例肛门生殖器肿瘤患者的样本作为对照和比较。在 53 例 HNC 患者的样本中，12 例（26.7%）HPV 阳性，33 例（73.3%）HPV 阴性，8 例（15.1%）Cobas 检测无结果。在 26 例肛门生殖器肿瘤患者的样本中，15 例（57.7%）HPV 阳性，11 例 HPV 阴性（42.3%）。一个样本发现同时感染 HPV 16 和 HPV 18。只有 3 个样本的结果不一致。Cobas 检测法适用于 HPV 的常规检测，对所有 HPV 基因型均具有极好的重复性和重现性（CV<0.6%和<0.4%）。Cobas 检测的敏感性和特异性分别为 91.7%[61.5%；99.8%]和 96.9%[83.8%；99.9%]。10ng DNA 足以检测 FFPE 和口腔刷样本中的 HPV 16、HPV 18 和 HPV。Cobas 检测与传统 PCR 具有可比性，可快速准确地检测 FFPE 和口腔刷样本中的 HPV DNA，用于 HNC 和其他类型 HPV 相关肿瘤的管理。
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