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The change of APP mRNA levels in peripheral blood of Alzheimer's disease patients using fluorescence quantitative PCR method
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- Author:
- No author available
- Journal Title:
- CHINESE JOURNAL OF NEUROMEDICINE
- Issue:
- 4
- DOI:
- 10.3760/cma.j.issn.1671-8925.2006.04.001
- Key Word:
- 阿尔茨海默病;淀粉样蛋白前体;荧光定量PCR;基因表达
Abstract： 目的建立荧光定量PCR方法检测Alzheimer病(AD)患者外周血中淀粉样蛋白前体(APP)的mRNA水平,并探讨该基因在AD患者外周血中的表达及意义.方法根据APP的基因序列,设计并合成引物和荧光标记探针.将PCR扩增目的片段用AT克隆的方法克隆入T载体,重组质粒经筛选、鉴定后,作为阳性模板,用于标准曲线的制定和样品检测.用该方法检测30例AD患者和23例正常老年人对照组外周血中APP基因的mRNA水平.结果应用重组质粒制作的定量曲线循环阈值与模板浓度具有良好的线性关系.AD组APP基因平均表达水平为(2.54×105±1.53×105)copies/μgRNA,高于对照组(6.03×104±7.58×105)copies/μgRNA(P＜0.001). 结论荧光定量PCR检测AD患者APPmRNA水平的方法较常规PCR技术更为简便、快速、准确.用该法测得APP在AD患者外周血中的mRNA水平有所增加.
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