利用36B4基因扩增效率评价PBMC基因组DNA的制备质量  

Evaluation of the preparation quality of genomic DNA from PBMC by using 36B4 gene amplification efficiency

作　　者：周丹[1] 崔晓娴 孙淑惠[3] 蒋琳[1] 荣星喻 汪慧菁[2] 赵超[2,3] 史虹莉[1] 

机构地区：[1]复旦大学附属华山医院健康管理中心,上海201206 [2]复旦大学老年医学研究中心和上海市老年医学临床重点实验室,上海200032 [3]复旦大学基础医学院教育部/卫计委医学分子病毒学重点实验室,上海200032

出　　处：《中国当代医药》2016年第28期4-7,共4页China Modern Medicine

基　　金：教育部霍英东教育基金会第十四届高等院校青年教师基金应用研究课题资助项目(142016);上海市老年医学临床重点实验室开放课题(13dz2260700)

摘　　要：目的建立一种评价PBMC基因组DNA制备质量的简便有效的方法。方法本研究采用实时定量PCR技术,对样本中的单拷贝基因36B4进行扩增,以考察扩增效率与样品质量之间的关系。结果经DNaseⅠ处理的基因组DNA中的36B4扩增效率显著低于未处理样品。采用该方法,对18份健康志愿者的血液标本基因组样品进行检测分析,结果显示,其可以有效地评估血液标本基因组制备质量。结论该方法可用于评价健康体检标本采集中的流程和操作对体检的影响,提高涉及基因组DNA检测体检项目的准确性。Objective To establish a simple and effective method for evaluating the preparation quality of the genomic DNA from PBMC.Methods In this study,real-time quantitative PCR technique was used to amplify the single copy gene 36B4 in a sample to investigate the relationship between the amplification efficiency and the quality of samples. Results The amplification efficiency of 36B4 in the genomic DNA treated by DNase [ was significantly lower than that of the untreated sample.By using this method,the blood samples from 18 healthy volunteers were detected and analyzed,the results showed that the preparation quality of blood sample genome was effectively evaluated.Conclusion The method can be used to evaluate the influence of the process and operation on the physical examination in the sample collection of health examination,and to improve the accuracy of the test item of the genomic DNA.

关 键 词：单拷贝基因 基因组DNA 血液标本 定量PCR 

分 类 号：R-331[医药卫生]