载脂蛋白E基因多态性检测技术的对比性研究

A contrast study of detection technology about ApoE gene polymorphism

目的建立并比较载脂蛋白Ｅ（ＡｐｏＥ）基因多态性检测技术，为ＡｐｏＥ基因分型提供良好的方法。方法应用聚合酶链反应（ＰＣＲ）限制性片段长度多态性（ＲＦＬＰ）和ＰＣＲ单链构象多态性（ＳＳＣＰ）检测技术对１１０例患者及相同人数的正常人进行ＡｐｏＥ基因型检测和分析。结果应用ＲＦＬＰ、ＳＳＣＰ技术测得正常对照组：ε２／２（０，０．０２０），ε３／３（０．７００，０．７６０），ε４／４（０，０．０４０），ε２／３（０．１００，０．０６０），ε２／４（０，０．０２０），ε３／４（０．２００，０．１００），ε２（０．０５０，０．０６０），ε３（０．８５０，０．８４０），ε４（０．１００，０．１００）。经χ２检验，ＡｐｏＥ基因型频率和等位基因频率的差异无显著意义（Ｐ＞０．０５）。上两种检测技术与既往两种检测技术（ＩＥＦ，ＷｅｓｔｅｒｐＢｌｏｔｉｎｇ）的结果经χ２检验，其ＡｐｏＥ等位基因频率的差异也无显著意义（Ｐ＞０．０５），均符合中国汉族人ＡｐｏＥ基因频率分布。结论ＰＣＲＲＦＬＰ方法是一种稳定可靠的ＡｐｏＥ基因多态性分析方法，多重ＰＣＲＳＳＣＰ技术更加简便有效，使ＡｐｏＥ基因多态性的研究更加完善。

Objective To establish and compare polymorphism testing methods of apolipoprotein E (ApoE) gene. Methods 110 patients and 110 normal persons were tested by using PCR RFLP and PCR SSCP testing method. The documents about the method of ApoE protein type and other gene types in normal persons were reviewed. Results No sigificant difference was observed between ApoE gene frequency and unit gene frequency by using RFLP and SSCP: ε 2/2 (0,0.020),ε 3/3 (0.700,0.760), ε 4/4 (0,0.040), ε 2/3 (0.100,0.060), ε 2/4 (0, 0.020), ε 3/4 (0.200,0.100), ε 2(0.050,0.060), ε 3(0.850,0.840), ε 4(0.100,0.100) being checked by using χ 2 method ( P >0.05). Also no significant difference was noted between ApoE gene frequency and unit gene frequency by using the two methods which were used in the past (IEF, Westerp blotting) and checked by χ 2 method ( P >0.05). They all agreed to the Chinese ApoE gene frequency distribution. Conclusion PCR RFLP method is a stable and reliable ApoE gene polymorphism method. PCR SSCP, simple and effective, makes the ApoE gene polymorphism measurement better.