Direct bisulphite conversion of cervical samples for DNA methylation analysis. Issue 10 (3rd October 2022)

• Record Type: Journal Article
• Authors: Verhoef, Lisanne; Floore, A. N; Doorn, Saskia; Cuschieri, Kate; Bhatia, Ramya; Hesselink, A. T; Meijer, Chris J.L.M.; Steenbergen, Renske D.M.; Heideman, Daniëlle A.M.
• Is Part Of: Epigenetics. Volume 17:Issue 10(2022)
• Page Start: 1173
• Page End: 1179
• Publication Date: 2022-10-03
• Subjects: Sodium bisulphite treatment -- high-throughput -- host-cell DNA methylation -- HPV triage -- cervical screening -- liquid-based cytology; Epigenesis -- Periodicals; Epigenetica; 572.86505
• DOI: 10.1080/15592294.2021.1992911
• Languages: English
• ISSNs: 1559-2294

Abstract

ABSTRACT: Sodium bisulphite conversion of DNA to separate methylated from unmethylated cytosines is a standard for methylation analysis. This study evaluated a direct cell conversion protocol on cervical samples as alternative to isolated genomic DNA as input. Clinician-collected cervical samples (n = 120) were subjected to a direct conversion protocol, or genomic DNA was isolated with a fixed amount used for subsequent bisulphite conversion. Converted samples were compared for ACTB control gene and methylation of FAM19A4 and miR124-2 genes using quantitative methylation-specific PCR (QIAsure Methylation Test). Direct conversion resulted in a high success rate, i.e., 119/120 (99.2%) samples reported a valid test result. ΔΔCq values of FAM19A4 and miR124-2 were significantly correlated between both protocols (Spearman Rho 0.708 and 0.763, respectively, all p-values = 0.000). Agreement between both the bisulphite protocols was demonstrated by Bland–Altman plots. A direct cell conversion protocol shows good technical and analytical performance and offers a streamlined workflow for methylation analysis.