改良全血DNA 提取法的质量与效度分析

doi:10.16389/j.cnki.cn42-1737/n.2020.05.005

江汉大学学报（自然科学版） ›› 2020, Vol. 48 ›› Issue (5): 42-46.doi: 10.16389/j.cnki.cn42-1737/n.2020.05.005

改良全血DNA 提取法的质量与效度分析

左文博¹，王嘉政¹，徐颖¹，杨小婷¹，倪维²，胡松^*1

1. 江汉大学医学院，湖北武汉 430056；2. 湖北省中医院，湖北武汉 430061

发布日期:2020-09-22
作者简介:左文博（1997— ），男，研究方向：免疫学。
基金资助:
HU Song

Quality and Validity Analysis of Improved Whole Blood DNA Extraction Method

ZUO Wenbo¹，WANG Jiazheng¹，XU Ying¹，YANG Xiaoting¹，NI Wei²，HU Song^*1

1. School of Medicine，Jianghan University，Wuhan 430056，Hubei，China；2. Hubei Provincial Hospital of TCM，Wuhan 430061，Hubei，China

Published:2020-09-22
Contact: 胡松
Supported by:
国家自然科学基金资助项目（31470270）；江汉大学2018 年度学生学术科技项目（2018yb249）

摘要/Abstract

摘要： 目的 为适应大批量提取外周血DNA 需求，针对现有试剂盒提取法进行改良。方法 将武汉地区健康人群血液样本80 份，随机分为两组，每组40 份。分别采用改良全血DNA 提取法和外周血单个核细胞（peripheral blood mononuclear cell，PBMC）DNA 提取法提取DNA，比较其效度和质量。结果 通过比较分析实验时间、提取DNA 的质量、纯度和PCR 结果，发现改良全血DNA 提取法提取的时间显著少于全血PBMC DNA 提取法，提取DNA 质量显著高于全血PBMC DNA 提取法（P <= 0. 05），而两种方法提取的基因组DNA 的OD₂₆₀/OD₂₈₀ 数值差异无统计学意义（P > 0. 05），且PCR 产物均可见单一且明亮的条带。结论 本实验中改良全血DNA 提取法是一种快速、经济、高效的DNA 提取方法，可在临床移植配型、临床基因多样性检测中发挥作用。

关键词: 改良全血DNA 提取法, 全血PBMC DNA 提取法, 基因组DNA

Abstract: Objective To meet the demand for large-scale extraction of peripheral blood DNA，we make improvements in the existing kit extraction method. Methods Eighty blood samples from healthy people in Wuhan were randomly divided into two groups with 40 samples in each group. Improved whole blood DNA extraction method and PBMC（peripheral blood mononuclear cell） DNA extraction method were used to extract DNA respectively， and their validity and quality were compared. Results By comparing and analyzing the experimental time，the quality and purity of the extracted DNA，and the PCR results， it was found that the improved whole blood DNA extraction method took significantly less time than the whole blood PBMC DNA extraction method， and the extracted DNA quality was significantly higher than the whole blood PBMC DNA extraction method（P < 0. 05），while the OD260/OD280 values of the genomic DNA extracted by the two methods were not statistically different（P > 0. 05），and single and bright bands can be seen in the PCR products of the two methods. Conclusion The improved whole blood DNA extraction method is a fast，economical，and efficient DNA extraction method，which can play a role in clinical transplantation matching and clinical genetic diversity detection.

Key words: improved whole blood DNA extraction method, PBMC DNA extraction method from whole blood, genomic DNA

中图分类号:

R446.61

左文博，王嘉政，徐颖，杨小婷，倪维，胡松. 改良全血DNA 提取法的质量与效度分析[J]. 江汉大学学报（自然科学版）, 2020, 48(5): 42-46.

ZUO Wenbo，WANG Jiazheng，XU Ying，YANG Xiaoting，NI Wei，HU Song. Quality and Validity Analysis of Improved Whole Blood DNA Extraction Method[J]. Journal of Jianghan University (Natural Science Edition), 2020, 48(5): 42-46.

改良全血DNA 提取法的质量与效度分析

Quality and Validity Analysis of Improved Whole Blood DNA Extraction Method

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