法医学杂志 ›› 2012, Vol. 28 ›› Issue (1): 41-43.DOI: 10.3969/j.issn.1004-5619.2012.01.010

• 论著 • 上一篇    下一篇

简并寡核苷酸引物PCR技术的建立及其检测灵敏度分析

邓建强1,刘宝琴2,蔡继峰3,李文慧1,龙  仁1,侯一平4   

  1. (1. 海南医学院法医鉴定中心,海南 海口 570102; 2. 西安现代妇产医院,陕西 西安 721000; 3. 中南大学湘雅医学院法医学系,湖南 长沙 410078; 4. 四川大学华西基础医学与法医学院,四川 成都 610041)
  • 发布日期:2012-02-25 出版日期:2012-02-28
  • 通讯作者: 侯一平,男,教授,主要从事法医物证学研究;E-mail:rechtsme@wcums.edu.cn
  • 作者简介:邓建强(1971—),男,陕西千阳人,博士,副主任法医师,主要从事法医物证学与法医病理学研究;E-mail:forensicpatho@163.com
  • 基金资助:

    国家自然科学基金资助项目(30271446);高校博士点专项科研基金资助项目(20020610044)

Degenerate Oligonucleotide Primed-PCR Technology Establishment and Its Sensitivity Test Analysis

DENG JIAN-QIANG1, LIU BAO-QIN2, CAI JI-FENG3, LI WEN-HUI1, LONG REN1, HOU YI-PING4   

  1. (1. Department of Forensic Medicine, Hainan Medical College, Haikou 570102, China; 2. Xi’an Modern Woman Health Hospital, Xi’an 721000, China; 3. Department of Forensic Medicine, Xiangya School of Medicine, Central South University, Changsha 410078, China; 4. West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China)
  • Online:2012-02-25 Published:2012-02-28

摘要: 目的 建立基于简并寡核苷酸引物PCR(degenerate oligonucleotide primed-PCR,DOP-PCR)的全基因组检测体系,并对其可靠性、灵敏度进行研究。 方法 通过荧光标记STR复合扩增毛细管电泳检测系统,对DOP-PCR扩增产物进行检测,获得DOP-PCR检测体系的灵敏度、可靠性。 结果 成功建立了可靠的DOP-PCR检测体系,经过DOP-PCR预处理再进行STR分型检验,其检测灵敏度可达到5个细胞量(相当于30 pg)。 结论 本研究建立的DOP-PCR技术可靠且可提高法医学微量检材的检测灵敏度。

关键词: 法医遗传学, 基因扩增, 简并寡核苷酸引物PCR

Abstract: Objective To establish a whole genome amplification testing system based on degenerate oligonucleotide primed-PCR(DOP-PCR) and to explore its reliability and sensitivity. Methods DOP-PCR amplified production was detected by fluorescent labeled multiplex STR amplification and capillary electrophoresis detection system to determine reliability and sensitivity of DOP-PCR system. Results DOP-PCR system was successfully established and the detection sensitivity reached 5 cells (30 pg) by pretreatment of DOP-PCR and then detection of STR genotyping. Conclusion The system established in this study is reliable and more testing sensitive for forensic trace evidence.

Key words: forensic genetics, gene amplification, degenerate oligonucleotide primed-PCR

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