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焦磷酸测序和MassARRAY技术用于年龄估计的DNA甲基化定量分析:一项比较研究

[Quantitative analysis of DNA methylation by pyrosequencing and MassARRAY technique for age estimation: a comparative study].

作者信息

Wang Ling, Peng Fuduan, Zhao Hui, Li Shanfei, Sun Xiaomeng, Liu Tianzi, Feng Lei

机构信息

Institute of Forensic Science, Ministry of Public Security//Ministry of Public Security Key Laboratory of Forensic Genetics, Beijing 100000, China.

China University of Political Science and Law, Institute of Evidence Law and Forensic Science, Beijing 100088.

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2020 Dec 30;40(12):1831-1837. doi: 10.12122/j.issn.1673-4254.2020.12.21.

Abstract

OBJECTIVE

To study the difference in age estimation based on quantitative analysis of DNA methylation by MassARRAY and pyrosequencing techniques.

METHODS

The methylation levels of 9 CpG sites from two independent whole blood sample sets (containing 65 and 62 samples) were detected using MassARRAY and pyrosequencing techniques. Z-score transformation was used to remove the batch effects of different techniques, and a linear regression model was used for age prediction.

RESULTS

For age prediction using the MassARRAY system, the 65 samples showed a mean absolute difference (MAD) of 2.49 years before Z-score transformation of the data and 2.44 years after the transformation, similar to the results in the 62 samples (MAD of 3.36 years before and 3.42 years after Z-score transformation). For data typed from pyrosequencing, the 65 samples showed a MAD of 4.20 years before and 2.76 years after data Z-score transformation, also similar to the results in the 62 samples (MAD of 3.92 years before and 3.63 years after data transformation).

CONCLUSIONS

Z-score transformation can effectively reduce the system batch effect between MassARRAY and pyrosequencing. Data from the MassARRAY system allows direct age estimation without further data processing, while the pyrosequencing data may increase the error in age estimation, which can be corrected by Z-score transformation of the data.

摘要

目的

通过MassARRAY和焦磷酸测序技术对DNA甲基化进行定量分析,研究年龄估计的差异。

方法

使用MassARRAY和焦磷酸测序技术检测两个独立全血样本集(分别包含65个和62个样本)中9个CpG位点的甲基化水平。采用Z分数转换去除不同技术的批次效应,并使用线性回归模型进行年龄预测。

结果

对于使用MassARRAY系统进行年龄预测,65个样本在数据进行Z分数转换前的平均绝对差(MAD)为2.49岁,转换后的平均绝对差为2.44岁,与62个样本的结果相似(Z分数转换前为3.36岁,转换后为3.42岁)。对于焦磷酸测序分型的数据,65个样本在数据进行Z分数转换前的平均绝对差为4.20岁,转换后的平均绝对差为2.76岁,也与62个样本的结果相似(数据转换前为3.92岁,转换后为3.63岁)。

结论

Z分数转换可有效降低MassARRAY和焦磷酸测序之间的系统批次效应。MassARRAY系统的数据无需进一步数据处理即可直接进行年龄估计,而焦磷酸测序数据可能会增加年龄估计的误差,通过对数据进行Z分数转换可对其进行校正。

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