法医学杂志

法医学杂志 ›› 2010, Vol. 26 ›› Issue (5): 349-352.DOI: 10.3969/j.issn.1004-5619.2010.05.008

• 论著 • 上一篇    下一篇

HE染色切片组织的STR检测及法医学应用

柳  燕，赵珍敏，李  莉，邓恺飞   

  1. （司法部司法鉴定科学技术研究所 上海市法医学重点实验室，上海 200063）
  • 发布日期:2010-10-25 出版日期:2010-10-28
  • 作者简介:柳燕（1963—），女，云南昆明人，副研究员，主要从事法医遗传学研究；E-mail：qliuyan@163.com
  • 基金资助:

    中央级科研院所社会公益研究资助项目（GY0804）

STR Profiling of Genomic DNA from HE Stained Tissue Sections

LIU YAN, ZHAO ZHEN-MIN, LI LI, DENG KAI-FEI   

  1. （Shanghai Key Laboratory of Forensic Medicine, Institute of Forensic Science, Ministry of Justice, P.R.China, Shanghai 200063, China）
  • Online:2010-10-25 Published:2010-10-28

被引次数

5

摘要： 目的 建立并优化HE染色切片组织STR分型方法，评估HE染色切片组织的STR分型有效性。 方法 用QIAgen法提取2例尸检人体的心、肝、肺、肠等8种组织的HE染色切片DNA，用TaqMan荧光探针法通过荧光域值循环数（Ct值）测定获得各提取液中的DNA质量浓度，以阳性内对照（internal positive control，IPC）监测PCR过程中的抑制水平。再用Identifiler试剂盒扩增，在3100-Avant上进行STR片段分析。 结果 在本研究建立优化的STR分型技术条件下，8种人体组织的HE染色切片DNA提取液质量浓度均可达到1 ng/μL以上，其IPC的Ct值提示无PCR抑制剂。HE染色切片组织的STR分型有效性与石蜡包埋组织相当，其DNA随时间延续而缓慢降解。 结论 在一定保存时限内，HE染色切片的DNA质和量可以满足STR分型检测需要，但受残余甲醛固定剂的影响，HE染色切片的分型成功率随保存时间延长而降低。

关键词: 法医遗传学, 石蜡包埋, STR分型, HE染色

Abstract: Objective To develop a STR analysis method for analyzing DNA from stained tissue sections and to evaluate the capability of this protocol in forensic application. Methods Eight kinds of HE stained human tissue, for example heart, liver, lung and intestine, were collected from two autopsy cases. The genomic DNA from those tissues was extracted using a QIAgen kit. DNA quantitation was performed using the TaqMan PCR method. The concentration of DNA isolated was determined based on Ct values. Internal positive controls （IPC） were used to monitor inhibitors. DNA amplifications were performed using Identifiler PCR Amplification kit. PCR products were analyzed on 3100-Avant Genetic Analyzer. Results The concentrations of DNA obtained from all samples were greater than 1 ng/μL. PCR inhibition was not observed. However, DNA degradation, potentially due to the effect of residual formalin fixative, was observed among tissue samples stored for long periods of time. Conclusion Sufficient amounts of DNA were extracted from HE stained tissue sections. STR profiles were successfully generated. The number of genotype alleles detected decreased as sample storage time increased.

Key words: forensic genetics, paraffin embedding, STR typing, HE staining

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