法医学杂志 ›› 2023, Vol. 39 ›› Issue (1): 45-49.DOI: 10.12116/j.issn.1004-5619.2022.520202

• 技术与应用 • 上一篇    下一篇

细胞裂解法和磁珠法在法医DNA检验中的对比

施嘉骏1(), 吴丹2, 刘铁柱3, 郝思静1, 孟必成1, 李士林4, 刘亚楠1()   

  1. 1.法医物证学现场应用技术公安部重点实验室 上海市现场物证重点实验室 上海市刑事科学技术研究院,上海 200083
    2.上海公安学院,上海 200137
    3.上海市公安局普陀分局刑事科学技术研究所,上海 200333
    4.复旦大学生命科学学院人类学与人类遗传学系,上海 200438
  • 收稿日期:2022-02-24 发布日期:2023-02-25 出版日期:2023-02-28
  • 通讯作者: 刘亚楠
  • 作者简介:刘亚楠,男,硕士,副主任法医师,主要从事法医物证DNA分析技术的应用和研究;E-mail:dyndai@sina.cn
    施嘉骏(1992—),男,硕士,主检法医师,主要从事法医物证DNA分析技术的应用和研究;E-mail:357976470@qq.com
  • 基金资助:
    公安部科技强警基础工作专项资助项目(2020GABJC33)

Comparative of Forensic DNA Identification Using Cell Lysis Method and Magnetic Beads Method

Jia-jun SHI1(), Dan WU2, Tie-zhu LIU3, Si-jing HAO1, Bi-cheng MENG1, Shi-lin LI4, Ya-nan LIU1()   

  1. 1.Key Laboratory of Forensic Evidence and Science Technology, Ministry of Public Security, Shanghai Key Laboratory of Crime Scene Evidence, Shanghai Research Institute of Criminal Science and Technology, Shanghai 200083, China
    2.Shanghai Public Security College, Shanghai 200137, China
    3.Institute of Criminal Science and Technology, Putuo Branch of Shanghai Public Security Bureau, Shanghai 200333, China
    4.Department of Anthropology and Human Genetics, School of Life Sciences, Fudan University, Shanghai 200438, China
  • Received:2022-02-24 Online:2023-02-25 Published:2023-02-28
  • Contact: Ya-nan LIU

摘要:

目的 比较细胞裂解法和磁珠法在法医DNA检验中的效果,探讨两种方法在法医学检验中的运用。 方法 使用细胞裂解法和磁珠法获取不同数量的THP-1细胞的基因组DNA并使用实时定量PCR检测DNA含量。使用细胞裂解法和磁珠法对不同稀释倍数的人血进行STR分型检测。 结果 当THP-1细胞数量为100、400和800个时,使用细胞裂解法提取的DNA含量分别为(1.219±0.334)、(5.081±0.335)、(9.332±0.318) ng;使用磁珠法提取的DNA含量分别为(1.020±0.281)、(3.634±0.482)、(7.896±0.759) ng,在THP-1细胞数量为400和800个时,细胞裂解法提取的DNA含量高于磁珠法(P<0.05)。使用细胞裂解法和磁珠法检测不同稀释倍数人血的STR分型时灵敏度相近,在血样稀释100、300和500倍时均能获取完整的STR分型,血样稀释700倍时,均无法检出完整STR分型。使用细胞裂解法无法检出未稀释人血的STR分型。 结论 细胞裂解法操作方便,能最大程度保留模板DNA,有望适用于微量血痕检验。

关键词: 法医遗传学, 细胞裂解法, 磁珠法, 短串联重复序列, THP-1细胞

Abstract:

Objective To compare the effects of cell lysis method and magnetic beads method in forensic DNA identification and to explore these two methods in forensic DNA identification. Methods The genome DNA of THP-1 cells in different quantities was extracted by the cell lysis method and magnetic beads method, and the DNA content was quantified by real-time quantitative PCR. The cell lysis method and magnetic beads method were used to type the STR of human blood with different dilution ratios. Results When the numbers of THP-1 cell were 100, 400 and 800, the DNA content extracted by cell lysis method were (1.219±0.334), (5.081±0.335), (9.332±0.318) ng, respectively; and the DNA content extracted by magnetic beads method were (1.020±0.281), (3.634±0.482), (7.896±0.759) ng, respectively. When the numbers of THP-1 cells were 400 and 800, the DNA content extracted by the cell lysis method was higher than that by the magnetic beads method. The sensitivity of cell lysis method and magnetic beads method was similar in STR typing of human blood at different dilution ratios. Complete STR typing could be obtained at 100, 300 and 500-fold dilutions of blood samples, but could not be detected at 700-fold dilution. STR typing of undiluted human blood could not be detected by cell lysis method. Conclusion The cell lysis method is easy to operate and can retain template DNA to the maximum extend. It is expected to be suitable for trace blood evidence tests.

Key words: forensic genetics, cell lysis method, magnetic beads method, short tandem repeat (STR), THP-1 cells

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