oncgnostics GmbH, Jena, Germany.
Institute of Medical Statistics, Information Sciences and Documentation, Jena University Hospital, Jena, Germany.
Clin Epigenetics. 2017 Oct 24;9:118. doi: 10.1186/s13148-017-0419-2. eCollection 2017.
HPV DNA testing as a primary screening marker is being implemented in several countries. Due to the high HPV prevalence in the screening population, effective triage strategies for HPV-positive cases are required. The aim of this study was to evaluate the performance of a methylation-specific real-time PCR assay (GynTect®) comprising six marker regions as a triage test.
An analytical sensitivity of 0.1 ng genomic DNA corresponding to 15 SiHa cells was achieved. Absolute specificity was observed in the presence of 20 ng unmethylated genomic DNA. In a clinical setting, cervical scrapes of 306 women showing abnormal colposcopy were tested for cytology, HPV positivity, and the GynTect markers ASTN1, DLX1, ITGA4, RXFP3, SOX17, and ZNF671. Of all women, histopathological data were available. The overall sensitivity for GynTect to detect CIN3+ was 67.7% (95% CI 57.3%-77.1%) whereas sensitivity was significantly higher for women of age ≥ 30 years ( = 0.04). All cancer cases ( = 5) were detected by GynTect. The overall false positive rate (= 1-specificity) for women with no CIN was 17.4% (95% CI 12.5-23.1%), with a higher proportion among HPV-positive women (24.0%, 95% CI 16.0-33.6%). In a triage screening setting, where all women underwent HPV testing and the HPV positives in addition GynTect testing, the overall sensitivity would slightly decline but specificity would reach the maximum value of 88.7% (95% CI 83.7-92.6%).
The GynTect® assay is a robust easy to use assay with high analytical sensitivity and specificity. Moreover, the performance of the assay based on cervical scrapes provides further evidence for the usefulness of methylation markers to detect HPV-positive women with clinically relevant disease.
HPV DNA 检测作为一种主要的筛查标志物，正在多个国家实施。由于筛查人群中 HPV 的高患病率，需要有效的 HPV 阳性病例分流策略。本研究的目的是评估包含六个标记区域的甲基化特异性实时 PCR 检测（GynTect®）作为一种分流检测方法的性能。
实现了 0.1ng 基因组 DNA（相当于 15 个 SiHa 细胞）的分析灵敏度。在存在 20ng 未甲基化基因组 DNA 的情况下，观察到绝对特异性。在临床环境中，对 306 名出现异常阴道镜的女性的宫颈刮片进行细胞学检查、HPV 阳性检测以及 GynTect 标记物 ASTN1、DLX1、ITGA4、RXFP3、SOX17 和 ZNF671 检测。所有女性均有组织病理学数据。GynTect 检测 CIN3+的总体灵敏度为 67.7%（95%CI57.3%-77.1%），年龄≥30 岁的女性灵敏度显著更高（=0.04）。所有癌症病例（=5）均被 GynTect 检测到。无 CIN 女性的总假阳性率（=1 特异性）为 17.4%（95%CI12.5%-23.1%），HPV 阳性女性的比例更高（24.0%，95%CI16.0%-33.6%）。在一种筛查性筛选设置中，所有女性都接受 HPV 检测，HPV 阳性者除了接受 HPV 检测外，还接受 GynTect 检测，总体灵敏度会略有下降，但特异性会达到 88.7%（95%CI83.7%-92.6%）的最大值。
GynTect® 检测是一种强大的、易于使用的检测方法，具有高分析灵敏度和特异性。此外，基于宫颈刮片的检测结果进一步证明了甲基化标志物在检测 HPV 阳性且具有临床相关疾病的女性中的有用性。
