Performance of a DNA methylation marker panel using liquid-based cervical scrapes to detect cervical cancer and its precancerous stages

Abstract

BackgroundA change of cervical cancer screening algorithms to an HPV-based screening setting is discussed in many countries, due to higher sensitivity of HPV testing compared to cytology. Reliable triage methods are, however, an essential prerequisite in such a setting to avoid overtreatment and higher screening costs.ResultsIn this study, a series of cervical scrapes collected in PreservCyt liquid-based cytology (LBC) medium from women with cervical cancer (n = 5), cervical intraepithelial neoplasia grade 1–3 (n = 74), and normal cytology (n = 201; further n = 352 collected in SureThin®) were assessed for methylation of the marker regions ASTN1, DLX1, ITGA4, RXFP3, SOX17, and ZNF671 using the GynTect assay and compared to cobas® HPV and CINtec Plus® biomarker results. All samples from women with cervical cancer, 61.2% of CIN3, 44.4% of CIN2 and 20.0% of CIN1 cases were scored positive for the GynTect methylation assay. In contrast, all CIN, irrespective of severity grade, and carcinomas were positive by both, CINtec Plus and cobas HPV. The specificity of GynTect for CIN3+ was 94.6% compared to 69.9% for CINtec Plus and 82.6% for cobas HPV (all HPV types) and 90.6% for cobas HPV 16/18. DNA methylation analysis of this methylation marker panel (GynTect assay) in cervical scrapes consistently detects cervical cancer and the majority of CIN3 as well as a subset of CIN1/2 lesions. The detection rate among cytologically normal samples is extraordinarily low (1.5%).ConclusionGynTect shows excellent performance when using cervical scrape material collected in liquid-based cytology media, a prerequisite for employing such a test as a triage in screening programs. Compared to the other test systems used in this work, GynTect showed higher specificity while still detecting all cancer cases.

Current status of development of methylation biomarkers for in vitro diagnostic IVD applications

“…It targets methylation of the panel of six genes: ASTN1, DLX1, ITGA4, RXFP3, SOX17, and ZNF671, in the DNA extracted from cervical smear, using methylation-specific real-time PCR technology. The test was shown to detect cervical intraepithelial neoplasia above CIN3 (CIN3 + ) with a sensitivity of 64.8% and specificity of 94.6% [63].…”

Evaluation of a methylation classifier for predicting pre-cancer lesion among women with abnormal results between HPV16/18 and cytology

“…In a self-sampling study [29], DNA methylation testing for MAL and miR-124 showed a sensitivity of 70.5% for detecting CIN2+. Also, some methylation kits were tested, such as the QIAsure [10] (based on FAM19A4 and has-mir124-2) and GynTect assay [30] (based on ASTN1, DLX1, ITGA4, RXFP3, SOX17, and ZNF671). The QIAsure methylation based on FAM19A4 and has-mir124-2 showed a sensitivity of 70.5% and a specificity of 67.8% for detecting HSIL+.…”

The use of molecular markers for cervical screening of women living with HIV in South Africa

“…At this moment no commercial assays are available for these markers. Other commercially available assays, such as the GynTect assay [48], have not been evaluated in WLHIV yet.…”