法医学杂志

法医学杂志 ›› 2007, Vol. 0 ›› Issue (4): 286-289.

• 论文 • 上一篇    下一篇

混合斑中精子细胞分离及其DNA制备方法

李鑫;胡兰;冯雪飞;刘晓;   

  1. 中国人民公安大学,公安部物证鉴定中心,北京市公安局刑侦总队,上海艾本德生物技术国际贸易有限公司 北京100038,淄博市公安局,山东淄博255000,北京100038,北京100085,上海200122
  • 发布日期:2007-08-25 出版日期:2007-08-28

Isolating Sperm Cells and Preparing DNA from Mixture Sample

LI XIN1,2,HU LAN3,FENG XUE-FEI4,LIU XIAO5(1.CHINESE PEOPLES PUBLIC SECURITY UNIVERSITY,BEIJING 100038,CHINA;2.ZIBO PUBLIC SECURITY BUREAU,ZIBO 255000,CHINA;3.INSTITUTE OF FORENSIC SCIENCE,MINISTRY OF PUBLIC SECURITY,BEIJING 100038,CHINA;4.FORENSIC MEDICAL   

  • Online:2007-08-25 Published:2007-08-28

被引次数

12

摘要： 目的尝试建立一种检测混合斑中精子细胞的方法。方法使用显微操作法捕获精子细胞,全基因组扩增(多重置换扩增)精子细胞DNA。结果对10管精斑检材的全基因组扩增,获得了高产、保真的产物。使用50μL体系对20个精子细胞直接进行全基因组扩增,省去了对起始模板的纯化过程,DNA扩增倍数达30000倍以上,片段长度大多在15 kb以上,其STRs复合扩增分型结果有可参照性。结论显微操作法可以有效捕获精子细胞,排除干扰,多重置换扩增可以提供足够量的产物用于法医DNA分析,该方法具有可行性。

关键词: 法医物证学, 显微操作法, 混合斑, 精子细胞, 多重置换扩增, 全基因组扩增, 短串联重复序列

Abstract: Objective To establish a method of preparing and analyzing DNA of sperm cells from sample mixtures.Methods Sperm cells were captured by micromanipulation method,DNA template was prepared from these sperm cells,and then amplified by multiple displacement amplification(MDA).Results High-yield of highly integrated DNA was produced by MDA from very small amount of DNA template of 20 sperm cells.The DNA could be amplified at least 30 000-fold,and the length of most the DNA segments were greater than 15kb,providing sufficient amount of short tandem repeat(STR) products for DNA genotyping.Conclusion The micromanipulation method is efficient to isolate sperm cells from sample mixtures and multiple displacement amplification can provide sufficient amount of DNA products for STR genotyping.The method seems to be feasible in detecting DNA from rape cases.

Key words: forensic physical evidence, micromanipulation method, sample mixtures, sperm cells, multiple displacement amplification, genome amplification, STR