中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2013, Vol. 8 ›› Issue (32): 3036-3046.doi: 10.3969/j.issn.1673-5374.2013.32.007

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

无创纳米磁珠法提取脑卒中患者唾液DNA

  

  • 收稿日期:2013-06-24 修回日期:2013-08-27 出版日期:2013-11-15 发布日期:2013-11-15
  • 基金资助:

    深圳市科技研发资金基础研究 (No. JC201105180757)；深圳市科技计划项目（No. 201002063）

A magnetic nanoparticles-based method for DNA extraction from the saliva of stroke patients

Li Yi1, Ying Huang1, Ting Wu2, Jun Wu3   

  1. Department of Neurology, Peking University Shenzhen Hospital, Shenzhen 518036, Guangdong Province, China
  • Received:2013-06-24 Revised:2013-08-27 Online:2013-11-15 Published:2013-11-15
  • Contact: Li Yi, M.D., Associate professor, Chief physician, Department of Neurology, Peking University Shenzhen Hospital, Shenzhen 518036, Guangdong Province, China, yilitj@hotmail.com
  • About author:Li Yi and Ying Huang contributed equally to this article.
  • Supported by:

    This work was supported by the Scientific and Technological Innovation Foundation of Shenzhen, China, No. JC201105180757 and Shenzhen Science and Technology Projects, No. 201002063.

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摘要：

亚甲四氢叶酸还原酶（MTHFR）C677T基因多态性是脑卒中的危险因素之一，对其进行基因筛查对脑卒中的预防有重要意义。实验采用纳米磁珠法提取70例脑卒中患者和70名健康对照者唾液标本中的DNA，并应用传统试剂盒法提取受试者血液中的DNA。Real-time PCR检测结果显示，唾液和血液样本中的MTHFR C677T基因分型结果完全一致，并与基因测序结果相吻合。脑卒中组的TT突变基因型频率显著高于对照组，且TT基因型受试者血浆同型半胱氨酸水平均较高。可见利用纳米磁珠从唾液样本中提取DNA可用于MTHFR C677T基因多态性的检测，实现了无创性基因筛查的目的。

关键词: 神经再生, 脑损伤, 脑卒中, 纳米磁珠, 唾液, 亚甲四氢叶酸还原酶, 同型半胱氨酸, 基因多态性, 基因筛查, 基金资助文章

Abstract:

C677T polymorphism in the methylenetetrahydrofolate reductase (MTHFR) gene is a risk factor for stroke, suggesting that widespread detection could help to prevent stroke. DNA from 70 stroke pa-tients and 70 healthy controls was extracted from saliva using a magnetic nanoparticles-based method and from blood using conventional methods. Real-time PCR results revealed that the C677T polymorphism was genotyped by PCR using DNA extracted from both saliva and blood samples. The genotype results were confirmed by gene sequencing, and results for saliva and blood samples were consistent. The mutation TT genotype frequency was significantly higher in the stroke group than in controls. Homocysteine levels were significantly higher than controls in both TT genotype groups. Therefore, this noninvasive magnetic nanoparticles-based method using saliva samples could be used to screen for the MTHFR C677T polymorphism in target populations.

Key words: neural regeneration, brain injury, stroke, magnetic nanoparticles, saliva, methylenetetrahydrofolate reductase, homocysteine, gene polymorphism, gene screening, grants-supported paper, neuroregeneration