法医学杂志

法医学杂志 ›› 2013, Vol. 29 ›› Issue (2): 125-126.DOI: 10.3969/j.issn.1004-5619.2013.02.012

• 技术与应用 • 上一篇    下一篇

循环次数与体系对DNA检测灵敏度的影响

张  璐1，2，王保捷1，丁  梅1，林子清2，庞  灏1，邢佳鑫1，宣金锋1   

  1. （1． 中国医科大学法医学院，辽宁 沈阳 110001； 2． 中国刑事警察学院法医学系，辽宁 沈阳 110035）
  • 发布日期:2013-04-25 出版日期:2013-04-28
  • 通讯作者: 王保捷，男，博士，教授，主要从事法医遗传学教学及研究；E-mail：bjwang@mail.cmu.edu.cn
  • 作者简介:张璐（1976—），女，吉林桦甸人，博士，副教授，主要从事法医遗传学研究；E-mail：zhangluzhaoyang@sohu.com

Influences of Cycle Index and Volume on Sensitivity of DNA Detection

ZHANG LU1,2, WANG BAO-JIE1, DING MEI1, LIN ZI-QING2, PANG HAO1, XING JIA-XIN1, XUAN JIN-FENG1   

  1. （1. School of Forensic Medicine, China Medical University, Shenyang 110001, China; 2. Department of Forensic Medicine, China Criminal Police University, Shenyang 100035, China）
  • Online:2013-04-25 Published:2013-04-28

被引次数

7

摘要： 目的 探讨通过增加PCR循环次数和缩小扩增体系改善DNA检测灵敏度的可行性。 方法 将10例无血缘关系健康志愿者精液DNA样本分别定量为50、40、30、25、20、15、10 pg/μL，分别用10、5、3 μL的体系和28、30、32、34、36次循环进行扩增，用3130遗传分析仪检测15个常染色体STR基因座。 结果 28次循环下，3 μL体系可对40 pg/μL及以上样本正确分型。20 pg/μL及以上样本在10、5、3 μL体系中，34次循环均可正确分型，增加到36次循环，出现非特异性谱带，无法正确分型。 结论 增加循环次数和缩小扩增体系在一定程度上可以提高DNA检测的灵敏度。

关键词: 法医遗传学, STR, 精液, 循环参数

Abstract: Objective To explore the feasibility of improving the sensitivity of DNA detection by increasing the PCR cycle index and decreasing the volume of amplifying system. Methods The DNA of semen were collected from 10 healthy irrelevant volunteers, and were quantified to 50, 40, 30, 25, 20, 15, 10 pg/μL, separately. All samples were then amplified in 10, 5, 3 μL volume and at 28, 30, 32, 34, 36 cycles, respectively. 3130 genetic analyzer was used to detect 15 autosomal STR loci. Results Under the situation of 28 cycles and 3 μL volume, samples which achieved >40 pg/μL could be correctly typed. Under the situation of 10, 5, 3 μL volume, samples which achieved >20 pg/μL could be correctly typed at 34 cycles. When increasing the index to 36 cycles, they could not be correctly typed because of the non-specific band. Conclusion DNA detecting sensitivity can be improved to a certain extent by increasing the cycle index and decreasing the volume of amplifying system.

Key words: forensic genetic, STR, semen, cycle index

中图分类号: