法医学杂志

法医学杂志 ›› 2016, Vol. 32 ›› Issue (3): 180-183.DOI: 10.3969/j.issn.1004-5619.2016.03.005

• 论著 • 上一篇    下一篇

24个Y-STR基因座荧光标记复合扩增体系的建立

刘  宏1，李  越1，刘长晖1，刘  超1，葛斌文2，陈林丽2   

  1. （1. 广州市刑事科学技术研究所 广东省法医遗传学重点实验室，广东 广州 510030； 2. 无锡中德美联生物科技有限公司，江苏 无锡 214174）
  • 发布日期:2016-06-25 出版日期:2016-06-28
  • 通讯作者: 刘超，男，博士，主任法医师，博士研究生导师，主要从事法医遗传学研究；E-mail：liuchaogzf@163.com
  • 作者简介:刘宏（1979—），男，硕士，副主任法医师，主要从事法医DNA检验和研究；E-mail：79liuhong@163.com
  • 基金资助:

    广东省法医遗传学重点实验室资助项目（2010A06 0801001）；广东省科技计划资助项目（2013B021500010）

Development of a Fluorescence Multiplex Amplification System with 24 Y-STR Loci

LIU HONG1, LI YUE1, LIU CHANG-HUI1, LIU CHAO1, GE BIN-WEN2, CHEN LIN-LI2   

  1. （1. Guangdong Provincial Key Laboratory of Forensic Genetics, Guangzhou Institute of Forensic Science, Guangzhou 510030, China; 2. AGCU ScienTech Incorporation, Wuxi 214174, China）
  • Online:2016-06-25 Published:2016-06-28

被引次数

1

摘要： 目的 构建包含24个Y-STR基因座的荧光标记复合扩增体系。 方法 选择DYS531、DYS630、DYS622、DYS552、DYS510、DYS449、DYS459a/b、DYS446、DYS443、DYS635、DYS587、DYS527a/b、DYS460、Y-GATA-A10、DYS520、DYS557、DYS522、DYS481、DYS570、DYS385a/b、DYS444 24个Y-STR基因座，构建荧光复合扩增体系。检测该体系的特异性、同一性、灵敏度、扩增均衡性、抗干扰性和准确性，调查其在广东地区人群的基因多样性。 结果 建立的复合扩增体系在检测的非人类及女性样本中未发现谱带，同一个体不同组织检测结果一致，0.1 ng以上标准品9948可检测获得完整分型结果。常见抑制物中体系内加入120~200 μmol/L血红素、1.5~2.0 mmol/L钙离子时出现等位基因丢失，对靛蓝、腐殖酸、EDTA具有较强抗干扰性。通过146例无关个体与Yfiler系统平行检测比对，24 Y-STR检测体系分型准确。广东地区人群单倍型多样性（HD）为0.999 72，优于Yfiler系统（HD=0.998 58）。 结论 本研究建立的24个Y-STR基因座荧光标记复合扩增体系法医学应用前景广阔，可用于案件检验、亲权鉴定与Y-STR数据库建设。

关键词: 法医遗传学, Y染色体, 短串联重复序列, 荧光复合扩增

Abstract: Objective To establish a novel multiplex amplification system which comprises 24 Y-STR loci. Methods Total 24 Y-STR gene loci, concluding DYS531, DYS630, DYS622, DYS552, DYS510, DYS449, DYS459a/b, DYS446, DYS443, DYS635, DYS587, DYS527a/b, DYS460, Y-GATA-A10, DYS520, DYS557, DYS522, DYS481, DYS570, DYS385a/b, DYS444, were chosen for establishing the fluorescence multiplex amplification system. The specificity, identity, sensitivity, balance of the amplification, anti-interference and accuracy of the system were detected and the gene diversity was investigated in the population of Guangdong. Results No band was found in nonhuman and female samples that were tested by the established multiplex amplification system. The same genotyping results were obtained from different tissues of the same person. Complete profiles could be obtained from more than 0.1 ng of the standard sample 9948. The loss of alleles was found when the common inhibitors such as hemoglobin and calcium ion were added 120-200 μmol/L and 1.5-2.0 mmol/L respectively to the system which with a strong anti-interference to the indigo, humic acid and EDTA. The typing of 24 Y-STR system could give the reliable results when 146 unrelated male individuals were detected and compared with the Yfiler system parallelly. The haplotype diversity （HD） of the population in Guangdong reached 0.999 72 that was better than the result retained from Yfiler system, which the HD was 0.998 58. Conclusion The fluorescence amplification system with 24 Y-STR loci established in present study has a wildly application prospect and can be used for cases inspection, paternity tests and Y-STR database construction.

Key words: forensic genetics, Y chromatosome, tandem repeat sequences, fluorescent multiplex amplification

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