人外周血基因组DNA提取的影响因素及方案选择  

Influential Factor and Protocols Selection for Genomic DNA Extraction from Human Peripheral Blood Samples

作　　者：张振[1] 杨文涛 陈炯[1] 冯巍[1] 

机构地区：[1]河南科技大学法医学院,河南洛阳471003 [2]内乡县公安局,河南内乡474350

出　　处：《河南科技大学学报（医学版）》2016年第4期310-315,共6页Journal of Henan University of Science & Technology:Medical Science

基　　金：国家自然科学基金(81501634);河南科技大学高级别项目培育基金(012ZCX021)

摘　　要：目的人外周血是基因组DNA样品的主要来源之一,了解外周血基因组DNA提取的研究进展,有助于法医检案中选择并优化DNA提取方案。方法回顾外周血基因组DNA提取的主要环节和影响因素,综述文献进行比较研究。结果白细胞裂解是决定DNA提取产量和质量的关键环节,去污剂是各种白细胞裂解液的核心组分。裂解后DNA分离纯化是影响DNA提取的另一重要环节,目前固相介质分离纯化法应用较广。实验室手工方案在一般实验室处理小批量样品时仍具有独特优势,试剂盒方案目前应用较为广泛,自动化提取系统尤其适用于大批量样品处理。然而,任何方案均不能同时满足DNA产量与质量、操作时间、安全性、费用等指标的同时最优化的要求。结论人外周血基因组DNA提取方案众多,需综合考虑DNA提取效能、操作时间、安全性、费用、设备和试剂易得性等多种因素,选择适宜的DNA提取方案。Objective Human peripheral blood samples were one of the main sources used to obtain genomic deoxyribonucleic acid （gDNA）. In order to select the optimal DNA extraction protocol on forensic identification, it would be helpful to investigate the research progress of DNA extraction. Methods There reviewed the common procedures adopted by most DNA extraction protocols, and highlighted the principles of its operation and impact factors by searching the representative studies from database. Results The lysis of white blood cell （WBC） was a key step to determine the yield and quality of extracted DNA. Detergents were usually the core component of a variety of WBC lysis solution. Another key step was the separation and purification of free DNA which usually involved some solid media. The manual protocols had still common alternatives for DNA extraction from peripheral blood samples, especially when a few samples were processed in small laboratories. A variety of DNA extraction kits had been widely used at present. Several automated systems for gDNA extraction were very suitable for treating the large quantities of blood samples. However, there was not enough scientific evidence to support one particular DNA extraction method from whole blood samples in terms of DNA yield and quality, time consumption, safety, and cost. Conclusion The optimal choice for gDNA extraction had still based on comprehensive consideration of multiple items, such as DNA extraction efficiency, time consumption, safety, cost, and availability of equipment and reagents from variety protocols.

关 键 词：外周血 基因组DNA DNA提取 

分 类 号：D919.2[医药卫生—法医学]