法医学杂志

法医学杂志 ›› 2014, Vol. 30 ›› Issue (3): 188-190.DOI: 10.3969/j.issn.1004-5619.2014.03.008

• 论著 • 上一篇    下一篇

氨基比林血痕预试验处理血痕后样本的DNA定量

朱传红1，郑道利1，倪尧志2，王海生3，宁  平1，方  慧1，刘  艳4   

  1. （1. 武汉市公安局刑事侦查局，湖北 武汉 430024； 2. 武汉金宏生物科技发展有限责任公司，湖北 武汉 430015； 3. 湖北省公安厅刑警总队，湖北 武汉 430070； 4. 华中科技大学同济医学院法医学系，湖北 武汉 430030）
  • 发布日期:2014-06-25 出版日期:2014-06-28
  • 通讯作者: 刘艳，女，博士研究生，主要从事法医病理学及毒理病理学研究；E-mail：liuyan@mail.hust.edu.cn
  • 作者简介:朱传红（1968—），男，湖北武汉人，博士，主要从事法医分子生物学研究、法医现场勘查学检验及研究；E-mail：zhuchh027@sina.com
  • 基金资助:

    公安部2009年应用创新项目课题（2009YYCXHBST061）

DNA Quantification of Blood Samples Pre-treated with Pyramidon

ZHU CHUAN-HONG1, ZHENG DAO-LI1, NI RAO-ZHI2, WANG HAI-SHENG3, NING PING1, FANG HUI1, LIU YAN4   

  1. （1. Department of Criminal Investigation, Wuhan Public Security Bureau, Wuhan 430024, China; 2. Wuhan Jin Hong Biological Technology Co., Ltd., Wuhan 430015, China; 3. Team of Criminal Police, Hubei Public Security Bureau, Wuhan 430070, China; 4. Department of Forensic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China）
  • Online:2014-06-25 Published:2014-06-28

被引次数

1

摘要： 目的 探讨氨基比林血痕预试验处理血痕后样本DNA含量的变化及对STR分型检测的影响。 方法 10名健康无关个体EDTA抗凝血液制成滤纸血痕，氨基比林血痕预试验检测，按试验后血样干燥保存时间分30 min、1 h、3 h、6 h、12 h、24 h共6个实验组，并采用磁珠法、QIAcube DNA纯化法、Chelex-100法三种方法提取样本DNA，应用荧光定量PCR检测样本DNA含量，PCR-STR荧光技术进行STR分型。 结果 提取方法相同时，氨基比林血痕预试验后血样随干燥保存时间的延长，样本DNA含量呈逐渐降低的趋势。保存时间相同时，不同DNA提取方法间，样本DNA含量差异也有统计学意义。90.56%样本均可获得16个STR基因座明确分型。 结论 氨基比林血痕预试验对血痕样本DNA有损伤，24 h内多可获有效STR分型。磁珠法提取样本DNA进行STR分型，效果最好。

关键词: 法医遗传学, 氨基比林预试验, 磁珠法, QIAcube DNA纯化法, Chelex-100法

Abstract: Objective To study DNA quantification and STR typing of samples pre-treated with pyramidon. Methods The blood samples of ten unrelated individuals were anticoagulated in EDTA. The blood stains were made on the filter paper. The experimental groups were divided into six groups in accordance with the storage time, 30 min, 1 h, 3 h, 6 h, 12 h and 24 h after pre-treated with pyramidon. DNA was extracted by three methods: magnetic bead-based extraction, QIAcube DNA purification method and Chelex-100 method. The quantification of DNA was made by fluorescent quantitative PCR. STR typing was detected by PCR-STR fluorescent technology. Results In the same DNA extraction method, the sample DNA decreased gradually with times after pre-treatment with pyramidon. In the same storage time, the DNA quantification in different extraction methods had significant differences. Sixteen loci DNA typing were detected in 90.56% of samples. Conclusion Pyramidon pre-treatment could cause DNA degradation, but effective STR typing can be achieved within 24 h. The magnetic bead-based extraction is the best method for STR profiling and DNA extraction.

Key words: forensic genetics, pyramidon preliminary test, magnetic bead-based extraction, QIAcube DNA purification method, Chelex-100 method

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