法医学杂志

法医学杂志 ›› 2013, Vol. 29 ›› Issue (1): 17-20.DOI: 10.3969/j.issn.1004-5619.2013.01.004

• 论著 • 上一篇    下一篇

多重置换扩增在病理切片DNA分型中的应用

张  越1，陈  阳1，杨元立2，李继周2，李朝品1   

  1. （1. 皖南医学院医学三系，安徽 芜湖 241002； 2. 公安部物证鉴定中心，北京 100038）
  • 发布日期:2013-02-25 出版日期:2013-02-28
  • 通讯作者: 陈阳，男，博士，副教授，硕士研究生导师，主要从事法医分子病理学研究；E-mail：chy5454@163.com
  • 作者简介:张越（1972—），男，安徽灵璧人，硕士，讲师，主要从事法医遗传学研究；E-mail：905970576@qq.com
  • 基金资助:

    国家自然科学基金资助项目（C080804）；安徽省自然科学基金资助项目（070413088）；安徽省高校青年教师科研资助计划项目（2007jq1176）

Applying Multiple Displacement Amplification to DNA Typing in the Pathological Section

ZHANG YUE1, CHEN YANG1, YANG YUAN-LI2, LI JI-ZHOU2, LI CHAO-PIN1   

  1. （1. The Third Medical Department, Wannan Medical College, Wuhu 241002, China; 2. Institute of Forensic Science, Ministry of Public Security, P.R.China, Beijing 100038, China）
  • Online:2013-02-25 Published:2013-02-28

摘要： 目的 探讨多重置换扩增（multiple displacement amplification，MDA）技术应用于法医学组织病理切片DNA分型的可行性。 方法 采用重复拉丁方设计实验，按保存时间、组织类型、尸源年龄3个因素分7个水平进行随机分组，共制作98例病理切片。硅珠法制备DNA模板，对照组直接采用AmpF?STR® IdentifilerTM试剂盒进行PCR扩增，实验组进行MDA后再进行扩增。以新鲜尸体组织的分型结果为标准，对比分析切片实验组与对照组的基因座检出数与检出率。 结果 各保存时间切片实验组与对照组的基因座检出率相比差异有统计学意义（P<0.01）；组织切片保存时间为360 d以内，实验组的16个基因座检出率均可达到95%以上。各组织类型间的差异具有统计学意义（P<0.01），而尸源年龄组间差异无统计学意义（P>0.01）。 结论 MDA可提高病理切片模板基因组量，相对降低PCR扩增抑制物浓度，减少等位基因脱扣现象，有效提高基因座检出率，MDA在法医切片DNA分型中具有运用价值，但应注意切片保存时间与组织类型等因素对分型结果的影响。

关键词: 法医遗传学, 冷冻切片, 多重置换扩增

Abstract: Objective To investigate the feasibility of applying multiple displacement amplification （MDA） to DNA typing in forensic pathological section. Methods Ninety-eight pieces of pathological sections were prepared in terms of 3 factors as the period of preservation, tissue types and death ages, and randomized into groups by Latin square by double 7-order design. Silicon bead method was used to extract the DNA template. Compared with the PCR amplification performed directly by AmpF?詛STR?誖IdentifilerTM kit in the control group, MDA was performed before amplification in the experimental group. Based on the samples from fresh autopsies as the standard genotypes, the number of detection and the detection rate were analyzed and compared between the experimental group and the control group. Results Between the control group and the experimental group, there was significantly statistical difference regarding the rate of DNA typing in each period of the tissue sections preserved （P<0.01）. The detection rate of the 16 loci in the experimental group was more than 95% when the period of the tissue sections were preserved within 360 d. There was significant difference in different tissue types （P<0.01）. But there was no significant difference in different death ages （P>0.01）. Conclusion MDA is efficacious in DNA typing of forensic pathological sections, for it can improve the DNA template quantification through abating the inhibiting factor’s concentration of PCR and reducing the rate of allele drop out （ADO）. However, the period of the sections preserved and tissue types would affect the results of genotyping by MDA.

Key words: forensic genetics, frozen sections, multiple displacement amplification

中图分类号: