中国神经再生研究(英文版) ›› 2014, Vol. 9 ›› Issue (1): 69-75.doi: 10.4103/1673-5374.125332

• 原著:退行性病与再生 • 上一篇    下一篇

无创快速法分析晚发型阿尔茨海默病载脂蛋白E基因多态性

  

  • 收稿日期:2013-12-14 出版日期:2014-01-05 发布日期:2014-01-05
  • 基金资助:

    深圳市科技研发基因基础计划项目(No. 201002063 and JC201105180757A)

A non-invasive, rapid method to genotype late-onset Alzheimer’s disease-related apolipoprotein E gene polymorphisms

Li Yi1, Ting Wu1, Wenyuan Luo1, Wen Zhou2,  Jun Wu1   

  1. 1 Department of Neurology, Peking University Shenzhen Hospital, Shenzhen, Guangdong Province, China
    2 Department of Radiology, Peking University Shenzhen Hospital, Shenzhen, Guangdong Province, China
  • Received:2013-12-14 Online:2014-01-05 Published:2014-01-05
  • Contact: Li Yi, M.D., Department of Neurology, Peking University Shenzhen Hospital, Shenzhen 518036, Guangdong Province, China, yilitj@hotmail.com.
  • Supported by:

    This work was supported by two grants from Science, Industry, Trade and Information Technology Commission of Shenzhen Municipality in China, grant No. 201002063, JC201105180757A.

摘要:

载脂蛋白E基因ε4被认为是阻碍晚发型阿尔茨海默病的危险因素。实验拟建立一种无创快速分析晚发型阿尔茨海默病相关载脂蛋白E基因多态性的方法。故应用纳米磁珠提取晚发性阿尔茨海默病患者及健康对照者的口腔拭子基因组DNA,用TaqMan-BHQ探针实时荧光PCR法对载脂蛋白E基因多态性进行分型,再通过基因测序的方法验证实时荧光PCR基因分型的准确性。结果显示,实时荧光PCR基因分型与基因测序的结果一致,说明此方法的可靠性。作者认为,这种无创、快速分析晚发型阿尔茨海默病相关载脂蛋白E基因多态性的方法适合大规模人群中分析载脂蛋白E基因ε4基因型与晚发型阿尔茨海默病的关系。

关键词: 神经再生, 神经退行性变, 晚发型阿尔茨海默病, 载脂蛋白E, 实时PCR, DNA测序, 危险因素, 等位基因, 基金资助文章

Abstract:

The apolipoprotein E gene ε4 allele is considered a negative factor for neural regeneration in late-onset Alzheimer’s disease cases. The aim of this study was to establish a non-invasive, rapid method to genotype apolipoprotein E gene polymorphisms. Genomic DNA from mouth swab specimens was extracted using magnetic nanoparticles, and genotyping was performed by real-time PCR using TaqMan-BHQ probes. Genotyping accuracy was validated by DNA sequencing. Our results demonstrate 100% correlation to DNA sequencing, indicating reliability of our protocol. Thus, the method we have developed for apolipoprotein E genotyping is accurate and reliable, and also suitable for genotyping large samples, which may help determine the role of the apolipoprotein E ε4 allele in neural regeneration in late-onset Alzheimer’s disease cases.

Key words: nerve regeneration, neurodegeneration, late-onset Alzheimer’s disease, apolipoprotein E gene, real-time PCR, DNA sequencing, risk factor, allele, neural regeneration