简并寡核苷酸引物PCR技术的建立及其检测灵敏度分析  

Degenerate Oligonucleotide Primed-PCR Technology Establishment and Its Sensitivity Test Analysis

作　　者：邓建强[1] 刘宝琴 蔡继峰[3] 李文慧[1] 龙仁[1] 侯一平[4] 

机构地区：[1]海南医学院法医鉴定中心,海南海口570102 [2]西安现代妇产医院,陕西西安721000 [3]中南大学湘雅医学院法医学系,湖南长沙410078 [4]四川大学华西基础医学与法医学院,四川成都610041

出　　处：《法医学杂志》2012年第1期41-43,共3页Journal of Forensic Medicine

基　　金：国家自然科学基金资助项目(30271446);高校博士点专项科研基金资助项目(20020610044)

摘　　要：目的建立基于简并寡核苷酸引物PCR(degenerate oligonucleotide primed-PCR,DOP-PCR)的全基因组检测体系,并对其可靠性、灵敏度进行研究。方法通过荧光标记STR复合扩增毛细管电泳检测系统,对DOP-PCR扩增产物进行检测,获得DOP-PCR检测体系的灵敏度、可靠性。结果成功建立了可靠的DOP-PCR检测体系,经过DOP-PCR预处理再进行STR分型检验,其检测灵敏度可达到5个细胞量(相当于30 pg)。结论本研究建立的DOP-PCR技术可靠且可提高法医学微量检材的检测灵敏度。Objective To establish a whole genome amplification testing system based on degenerate oligonucleotide primed-PCR（DOP-PCR） and to explore its reliability and sensitivity.Methods DOP-PCR amplified production was detected by fluorescent labeled multiplex STR amplification and capillary electrophoresis detection system to determine reliability and sensitivity of DOP-PCR system.Results DOP-PCR system was successfully established and the detection sensitivity reached 5 cells（30 pg） by pretreatment of DOP-PCR and then detection of STR genotyping.Conclusion The system established in this study is reliable and more testing sensitive for forensic trace evidence.

关 键 词：法医遗传学 基因扩增 简并寡核苷酸引物PCR 

分 类 号：DF795.2[医药卫生—法医学]