载脂蛋白E基因型分型方法研究

目的：为调查海南省老年性痴呆患者载脂蛋白Ｅ（Ａｐｏ Ｅ）基因频率，建立一种简便、快速、准确的人载脂蛋白Ｅ基因型检测方法。方法：应用碘化钠法提取模板ＤＮＡ，聚合酶链反应（ＰＣＲ）特异性扩增Ａｐｏ Ｅ基因含１１２ 位和１５８ 位氨基酸的基因序列，扩增产物用限制内切酶Ｈｈａ Ｉ酶切，聚丙烯酰胺凝胶电泳后，观察酶切位点的限制性片段长度多态性（ＲＦＩＰ）图谱。结果：碘化钠法提取模板ＤＮＡ 与经典法无差异；运用ＰＣＲ－ ＲＦＩＰ法检测３０ 名健康人Ａｐｏ Ｅ基因型，实验成功。结论：该方法简单、快速、准确，适合于一般实验室开展Ａｐｏ Ｅ基因型调查

METHODOLOGY FOR APOLIPOPROTEIN E GENOTYPING

Objective: To establish a simple,immediate and accurate method for apolipoprotein E(Apo E) genotyping in patients with Alzheimer's dementia in Hainan Province. Method: The template DNA was collected with the NaI method. Apo E gene sequences containing amino acid positions 112 and 158 were amplified for electrophoresis on polyacrylamide gels by polymerase chain reaction(PCR). The patterns of restriction fragment length polymorphism of Apo E gene were distinguished. Results:The NaI method showed no difference as compared with the classical method for collecting template DNA. Thus, Apo E genotyping by PCRRfip was carried out with success in 30 healthy individuals. Conclusion: The method is considered to be simple, immediate, and accurate for Apo E genotyping in ordinary laboratories.