[聚合酶链反应结合银染法用于载脂蛋白E等位基因的测定]
[PCR in combination with silver as a method for the determination of apolipoprotein E alleles].
作者信息
Zhang A, Han Z, Wang L, Tao G
机构信息
Institute of Geriatrics, PLA General Hospital, Beijing 100853 P. R. China.
出版信息
Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 1998 Oct 10;15(5):310-2.
OBJECTIVE
To present a rapid, sensitive and safe procedure for the determination of human apolipoprotein E alleles.
METHODS
Genomic DNA was extracted from 0.5ml whole blood by a non-phenol protocol. After PCR and restriction enzyme digestion,short DNA fragments were detected by a simplified silver staining method. This technique was used to assay apolipoprotein E alleles in 19 patients with sporadic Alzheimer's disease and 41 normal aged persons.
RESULTS
The short DNA fragments were visualized clearly on polyacrylamide gel processed by silver staining. It took 24 hours from DNA extraction to the final result. The A4 allele frequency in patients with sporadic Alzheimer's disease was 0.29, much higher than that in control group (0.02, P<0.001).
CONCLUSION
PCR in combination with silver staining can satisfy the need for high-resolution and high-efficiency in the determination of apolipoprotein E alleles and can be used as a routine procedure in clinical and epidemiological investigations.
目的
介绍一种快速、灵敏且安全的人类载脂蛋白E等位基因检测方法。
方法
采用非酚法从0.5ml全血中提取基因组DNA。经聚合酶链反应（PCR）和限制性内切酶消化后，用简化银染法检测短DNA片段。运用该技术对19例散发性阿尔茨海默病患者及41名正常老年人的载脂蛋白E等位基因进行检测。
结果
经银染处理的聚丙烯酰胺凝胶上可清晰观察到短DNA片段。从DNA提取到得出最终结果耗时24小时。散发性阿尔茨海默病患者中A4等位基因频率为0.29，远高于对照组（0.02，P<0.001）。
结论
PCR结合银染法能够满足载脂蛋白E等位基因检测中高分辨率和高效率的需求，可作为临床及流行病学调查的常规方法。
Zotero 插件