法医学杂志

法医学杂志 ›› 2018, Vol. 34 ›› Issue (6): 656-658.DOI: 10.12116/j.issn.1004-5619.2018.06.017

• 技术与应用 • 上一篇    下一篇

微量生物检材定量及分型

宋  立1，刘  松1，吴  卉1，方少平1，傅燕芳2   

  1. 1. 杭州市公安局西湖区分局刑事科学技术室，浙江 杭州 310013； 2. 浙江省公安物证鉴定中心 浙江省刑事科学技术应用研究重点实验室，浙江 杭州 310009
  • 发布日期:2018-12-25 出版日期:2018-12-28
  • 作者简介:宋立（1976—），男，副主任法医师，主要从事法医DNA鉴定；E-mail：songli05@163.com
  • 基金资助:
    浙江省刑事科学技术应用研究重点实验室开放基金资助项目（ZGWJ-KF2016009）

Quantification and Genotyping of Trace Samples

SONG Li1, LIU Song1, WU Hui1, FANG Shao-ping1, FU Yan-fang2   

  1. 1. Department of Criminal Science and Technology, Xihu District Branch of Hangzhou Public Security Bureau, Hangzhou 310013, China; 2. Zhejiang Key Laboratory of Forensic Science and Technology, Institute of Forensic Science, Zhejiang Province Public Security Bureau, Hangzhou 310009, China
  • Online:2018-12-25 Published:2018-12-28

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摘要： 目的 在传统微量生物检材提取方法中，引入荧光定量流程对样品进行初筛，以提高其有效检验。 方法 将DNA标准品9947A梯度稀释，使用Rotor-Gene Q实时荧光定量分析仪定量，用AmpF?STRTM IdentifilerTM Plus PCR扩增试剂盒检测基因型，构建微量生物检材可获得完整STR分型结果的定量阈值模型，并使用903个微量检材验证其可靠性。 结果 检材质量浓度>0.03 ng/μL可直接获得有效的分型；检材质量浓度>0.01~0.03 ng/μL，可通过优化增加PCR热循环参数（30个循环）获得有效分型；检材质量浓度≤0.01 ng/μL时，即使优化PCR也无法得到有效图谱。 结论 该实时荧光定量阈值模型可有效对微量生物检材进行初筛。

关键词: 法医遗传学, 串联重复序列, 实时荧光定量, 微量检材

Abstract: Objective To introduce real-time polymerase chain reaction （real-time PCR） into the initial sample screening, to improve the effectiveness of traditional trace sample extraction method. Methods Serial diluted 9947A was quantified using a Rotor-Gene Q real-time RT-PCR, and the genotype was determined with AmpF?STRTM IdentifilerTM Plus PCR kit. Thus a quantitative threshold model was built to obtain complete STR typing from the trace samples. In addition, 903 trace samples were used to verify the reliability. Results When the samples quality concentration was >0.03 ng/μL, the effective STR typing could be directly obtained; when the concentration was >0.01 and ≤0.03 ng/μL, the effective STR typing could be directly obtained by optimizing the PCR thermal cycle parameters （30 cycles）; and when the concentration was ≤0.01 ng/μL, no effective map could be obtained even if PCR was optimized. Conclusion The real-time PCR quantitative threshold model is effective for the screening of trace samples.forensic genetics; tandem repeat

Key words: forensic genetics, tandem repeat sequences, real-time fluorescence quantification, trace samples