基于下游实验的样本质量分析  

An Analysis of Sample Quality Based on Downstream Experiments

作　　者：刘丽[1,2] 林金嬉 梁宪红[1,2] 李上智 乔琳[1,2] 佟玲 秦海强 LIU Li;LIN Jinxi;LIANG Xianhong;LI Shangzhi;QIAO Lin;TONG Ling;QIN Haiqiang(BeijingTiantan Hospital,Capital Medical University,Beijing 100070,China;National Clinical Research Center for Neurological Diseases,Beijing 100070,China;Department of Neurology of BeijingTiantan Hospital,Capital Medical University,Beijing 100070,China)

机构地区：[1]首都医科大学附属北京天坛医院,北京100070 [2]国家神经系统疾病临床医学研究中心,北京100070 [3]首都医科大学附属北京天坛医院神经病学中心,北京100070

出　　处：《标记免疫分析与临床》2022年第11期1942-1945,共4页Labeled Immunoassays and Clinical Medicine

基　　金：国家自然科学基金资助项目(编号:81870907,82271299);“十三五”国家重点研发计划项目(编号:2016YFC1301604)。

摘　　要：目的依据ISO 20387生物样本库通用要求,通过下游实验对现有的白细胞提取质量进行验证,探索适用于生物样本库认可的样本前处理方法。方法对2021年1月至2022年6月使用白细胞进行靶向捕获测序的样本进行DNA提取,通过OD260/280、OD260/230、测量浓度,计算得率对DNA质量进行评价。结果5850例白细胞提取DNA,纯度足够高的样本占比为86.24%,得率大于10μg满足实验需求的样本占比为92.51%。结论将红细胞裂解步骤提前,在新鲜样本时进行,使用纯度较高的白细胞进行DNA提取,通过验证,上述提取方法可行,高纯度样本有效减少了红细胞的干扰,为样本库探索未知研究提供更为可靠的生物样本。Objective To verify the quality of the existing leukocyte extraction samples through downstream experiments according to the general requirements of ISO20387 biobanks,and to explore the sample pretreatment methods that are suitable for biobank approval.Methods From January,2021 to June,2022,DNA was extracted from samples using leukocytes by targeted capture sequencing,and the DNA quality was evaluated by OD260/280,OD260/230,measured concentration,and calculated yield.Results DNA was extracted from 5850 cases of leukocytes,in which the proportion of samples with sufficient purity was 86.24%,and the proportion of samples with a yield greater than 10ug to meet the experimental requirements was 92.51%.Conclusion Our results suggest that the red blood cell lysis step should be carried out in advance,and it is carried out in fresh samples,and DNA extraction with high purity white blood cells is helpful.After verification,this extraction method is feasible.The high purity samples can reduce the interference of red blood cells and provide the sample library for exploring unknown research.More reliable biological samples can be produced with this method.

关 键 词：质量 验证 DNA 

分 类 号：R393-33[医药卫生—基础医学]