法医学杂志

法医学杂志 ›› 2018, Vol. 34 ›› Issue (4): 401-404.DOI: 10.12116/j.issn.1004-5619.2018.04.012

• 技术与应用 • 上一篇    下一篇

镜下单片状头屑DNA提取分型法与EZ-tape法的比较

白小刚1，蹇  慧2，王  惠1，毛  炯1，夏  昱1，冯  涛1，陈  丹1，李庆庆1，朱  镜2，梁伟波2   

  1. 1. 成都市公安局，四川 成都 610081； 2. 四川大学华西基础医学与法医学院，四川 成都 610041
  • 发布日期:2018-08-25 出版日期:2018-08-28
  • 通讯作者: 朱镜，女，博士研究生，主要从事法医遗传学研究；E-mail：zhujingamy2012@163.com 梁伟波，男，副教授，硕士研究生导师，主要从事法医遗传学教学和科研；E-mail：liangweibo@scu.edu.cn
  • 作者简介:白小刚（1977—），男，主检法医师，主要从事法医遗传学鉴定；E-mail：435600590@qq.com
  • 基金资助:
    国家自然科学基金资助项目（81671871）

Comparison of Single Piece of Dandruff DNA Extraction under Microscope and EZ-tape Method

BAI Xiao-gang1, JIAN Hui2, WANG Hui1, MAO Jiong1, XIA Yu1, FENG Tao1, CHEN Dan1, LI Qing-qing1, ZHU Jing2, LIANG Wei-bo2   

  1. 1. Chengdu Public Security Bureau, Chengdu 610081, China; 2. West China School of Basic Science and Forensic Medicine, Sichuan University, Chengdu 610041, China
  • Online:2018-08-25 Published:2018-08-28

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摘要： 目的 改进常规头屑类样本的EZ-tape提取分型方法，借助显微镜采集单片状头屑，提高样本利用率、降低漏检率，并减少混合分型结果比例。 方法 采用EZ-tape脱落细胞粘取器和透明胶带粘取2名志愿者轮流佩戴的帽子内侧，EZ-tape法遵循传统方法直接提取DNA，镜下单片状头屑DNA提取分型法（简称“单片头屑分析法”）则借助显微镜分拣出透明胶带中的单片状头屑。两种方法均分别进行持续振荡和静置消化。所有样本均采用Chelex-100法提取DNA，扩增及电泳后获得STR分型。比较EZ-tape法及单片头屑分析法所获的STR分型结果。 结果 EZ-tape法所获分型中11份（45.8%）样本发生漏检，仅获得单一来源的分型结果，10份（41.7%）样本为混合分型结果，其中6份样本有等位基因插入或丢失。单片头屑分析法有12份样本获得了与志愿者一致的单一来源分型结果，仅2份样本获得混合分型结果。振荡处理的分型准确数高于静置消化（P<0.05）。 结论 DNA提取过程中进行振荡有利于DNA的释放。单片头屑分析法获得单一来源STR分型成功率较高，样本漏检率低，可作为法医临案工作中头屑类特殊检材的特定样本采集方式。

关键词: 法医遗传学, 头屑, EZ-tape, DNA提取

Abstract: Objective To collect single piece of dandruff with microscopes to improve the regular EZ-tape method for DNA extraction and genotyping, increase the utilization of samples, reduce the miss rate as well as the proportion of genotyping results of mixed stains. Methods The insides of the hats worn by two volunteers were stuck by EZ-tape and scotch tape respectively. DNA on EZ-tape was directly extracted using traditional method. Single piece of dandruff was collected from the scotch tapes under microscope. The two kinds of methods were both performed under continuous oscillation and standing digestion, respectively. DNA was extracted through Chelex-100 method, and STR genotypes were obtained after amplification and electrophoresis. The results of STR genotypes obtained by EZ-tape method and single piece of dandruff analytical method were compared. Results Miss detections happened in 11 samples （45.8%） by EZ-tape method and only single-source typing results were obtained. Ten samples （41.7%） showed the genotype results of mixed stain and six of which showed allele insertions and deletions. The genotype results were obtained successfully using the single piece of dandruff analytical method, and two samples showed mixed stain genotype. The number of exact typing processed by oscillation was higher than that by standing digestion （P<0.05）. Conclusion The oscillation during the DNA extraction process is in favour of the DNA releasing. Single piece of dandruff analytical method can be used to obtain single-source STR genotype with high successful ratio and low miss rate. This method can be a collection method of special samples such as dandruff in forensic practice.

Key words: forensic genetics, dandruff, EZ-tape, DNA extraction