法医学杂志 ›› 2011, Vol. 27 ›› Issue (1): 33-35,42.DOI: 10.3969/j.issn.1004-5619.2011.01.009

• 技术与应用 • 上一篇    下一篇

激光捕获显微切割技术用于分离混合斑中精子细胞

刘  芳1,王  静2,俞丽娟1,郭剑章1,高俊薇2,焦章平2,唐  晖2,刘雅诚2   

  1. (1. 山西医科大学 法医学院,山西 太原 030001; 2. 北京市公安局 法医检验鉴定中心,北京 100192)
  • 发布日期:2011-02-25 出版日期:2011-02-28
  • 通讯作者: 通信作者:王静,女,硕士,主要从事法医物证学研究;E-mail:wangj1118@126.com
  • 作者简介:刘芳(1985—),女,山西运城人,硕士研究生,主要从事法医物证学研究;E-mail:lf0911@qq.com

Isolating Sperm Cells by Laser Capture Microdissection Technique from Mixture Sample

LIU FANG1, WANG JING2, YU LI-JUAN1, GUO JIAN-ZHANG1, GAO JUN-WEI2, JIAO ZHANG-PING2, TANG HUI2, LIU YA-CHENG2   

  1. (1. School of Forensic Medicine, Shanxi Medical University, Taiyuan 030001, China; 2. Forensic Medical Examination Center, Beijing Public Security Bureau, Beijing 100192, China)
  • Online:2011-02-25 Published:2011-02-28

摘要: 目的 评估激光捕获显微切割(laser capture microdissection,LCM)技术在分离混合斑中少量精子细胞的应用价值。 方法 配制不同比例的精液-阴道上皮细胞混合液,分别用差异裂解法和LCM法分离精子细胞,用磁珠法提取精子细胞DNA,并用IdentifilerTM试剂盒进行STR基因型检测。 结果 LCM法分离精子细胞的STR成功率为92.86%(13/14),采用差异裂解法检测的成功率为7.14%(1/14),二者差异有统计学意义(P<0.05)。 结论 LCM法可有效排除女性细胞成分的干扰,并捕获少量精子细胞得到单一的男性STR分型,显著优于差异裂解法。

关键词: 法医遗传学, 精子细胞, 混合斑, 激光捕获显微切割技术

Abstract: Objective To assess the application value of laser capture microdissection(LCM) technique for isolating a small number of sperm cells from mixture sample. Methods Mixture samples were prepared with sperm cells and vaginal epithelia at different concentrations. Both LCM technique and the differential lysis method were employed to obtain sperm cells from the mixture samples, and DNA was extracted by magnetic beads method. STR genotyping was determined using IdentifilerTM kit. Results The successful STR genotype rate of sperm cells isolated from mixture samples with LCM technique was 92.86%(13/14). The rate of differential lysis method was 7.14%(1/14). The successful rates between the two methods were statistically different(P<0.05). Conclusion LCM technique can effectively exclude the interference of female cell component and isolate a small number of sperm cells to obtain a single male STR genotyping. LCM technique is obviously better than the differential lysis method.

Key words: forensic genetics, sperm cell, mixed stain, laser capture microdissection(LCM)

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