Department of Forensic Medicine, Seoul National University College of Medicine, Seoul, Korea.
Institute of Forensic and Anthropological Science, Seoul National University College of Medicine, Seoul, Korea.
Electrophoresis. 2021 Aug;42(14-15):1497-1506. doi: 10.1002/elps.202000312. Epub 2021 May 20.
DNA methylation is the most promising biomarker for estimating human age. There are various methods used for analyzing DNA methylation. Among those, the SNaPshot assay-based method provides a semi-quantitative measurement of DNA methylation using capillary electrophoresis on genetic analyzers. However, DNA methylation measures produced using different types of genetic analyzers have never been compared, although differences in methylation values can directly affect age estimates. To evaluate the differences between the results generated by different genetic analyzers, we analyzed the same blood, saliva, and control methylated DNA using three genetic analyzers-the Applied Biosystems 3130, 3500, and SeqStudio-and compared the methylation values at five CpG sites: ELOVL2, FHL2, KLF14, MIR29B2C, and TRIM59. The methylation value at each of the five CpG sites decreased in the order 3130, 3500, and SeqStudio. The differences in the results produced by the different genetic analyzers resulted in significant errors when applying the 3500 and SeqStudio data to a previous age estimation model constructed using the 3130 Genetic Analyzer data. Therefore, DNA methylation measurements from 3500 and SeqStudio were corrected using the regression functions obtained by plotting the DNA methylation data of one instrument versus the other to facilitate the application of DNA methylation data from one instrument to the age prediction model based on other instruments. The age prediction accuracy obtained by applying corrected 3500 and SeqStudio data to the existing age estimation model was as high as observed in the 3130 data.
DNA 甲基化是估计人类年龄最有前途的生物标志物。有各种分析 DNA 甲基化的方法。其中，SNaPshot 测定法基于毛细管电泳在基因分析仪上对 DNA 甲基化进行半定量测量。然而，使用不同类型的基因分析仪产生的 DNA 甲基化测量值从未进行过比较，尽管甲基化值的差异可能直接影响年龄估计。为了评估不同基因分析仪产生的结果之间的差异，我们使用三种基因分析仪——Applied Biosystems 3130、3500 和 SeqStudio——分析了相同的血液、唾液和对照甲基化 DNA，并比较了五个 CpG 位点的甲基化值：ELOVL2、FHL2、KLF14、MIR29B2C 和 TRIM59。五个 CpG 位点的甲基化值按 3130、3500 和 SeqStudio 的顺序降低。不同基因分析仪产生的结果差异导致将 3500 和 SeqStudio 数据应用于使用 3130 基因分析仪数据构建的先前年龄估计模型时产生显著误差。因此，使用回归函数校正了来自 3500 和 SeqStudio 的 DNA 甲基化测量值，该回归函数是通过绘制一个仪器的 DNA 甲基化数据与另一个仪器的 DNA 甲基化数据绘制得到的，以方便将一个仪器的 DNA 甲基化数据应用于基于其他仪器的年龄预测模型。将校正后的 3500 和 SeqStudio 数据应用于现有年龄估计模型时获得的年龄预测准确性与在 3130 数据中观察到的一样高。
