Amsterdam UMC, Vrije Universiteit Amsterdam, Pathology, Cancer Center Amsterdam, De Boelelaan Amsterdam, The Netherlands.
Self-screen B.v., Amsterdam, The Netherlands.
Epigenetics. 2022 Oct;17(10):1173-1179. doi: 10.1080/15592294.2021.1992911. Epub 2021 Oct 27.
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 control gene and methylation of and 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 and 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.
亚硫酸氢盐将 DNA 转化为分离甲基化和未甲基化胞嘧啶是甲基化分析的标准方法。本研究评估了一种直接细胞转化方案，作为替代分离基因组 DNA 作为输入的方法，用于宫颈样本。临床医生收集的宫颈样本（n=120）进行直接转化方案，或使用固定量的基因组 DNA 进行后续亚硫酸氢盐转化。使用定量甲基化特异性 PCR（QIAsure Methylation Test）比较转化样本中 和 基因的对照基因和甲基化。直接转化的成功率很高，即 119/120（99.2%）个样本报告了有效的测试结果。两种方案之间的 和 的 ΔΔCq 值显著相关（Spearman Rho 分别为 0.708 和 0.763，所有 p 值均=0.000）。Bland-Altman 图显示了两种亚硫酸氢盐方案之间的一致性。直接细胞转化方案显示出良好的技术和分析性能，并为甲基化分析提供了简化的工作流程。
