用热启动加两步改良PCR法探讨载脂蛋白E基因多态性与老年性骨质疏松的关系

Improve the efficiency and specificity of ApoE gene amplification by hot-started and two-step PCR,and search the relationship of ApoE gene type with senile osteoporosis

目的探索ApoE基因的优化扩增方法及ApoE基因多态性与老年人骨质疏松之间的关系。方法分别用常规PCR法和热启动加两步PCR法,对ApoE基因进行扩增,对比两种方法的扩增效果。收集60岁以上老年骨质疏松组和正常对照组患者血样各100份,采用热启动加两步PCR法对ApoE扩增产物进行测序,将测序结果与GENBANK中ApoE目的片段序列比对结果并分析ApoE基因多态性对骨质疏松影响。结果采用热启动加两步循环法优化ApoE的扩增方法,可以有效提高ApoE的PCR扩增的产率并可减少非特异性扩增。骨质疏松组E4等位基因频率高于对照组(P<0.05)。结论热启动加两步循环法可以有效地优化ApoE的扩增效率和特异性,ApoE4等位基因与老年性骨质疏松可能相关。

Objective To quest the method of improve the efficiency and specificity of ApoE gene amplification and the relationship between ApoE gene type and Senile Osteoporosis. Methods Compared Regulated PCR with Hot - started and two - step PCR in amplify ApoE gene. Subsequently, Compared Hot - started and two - step PCR sequence with GEN- BANK. Comparing the distribution of gene type and allelic gene of Senile osteoporosis group with control group. Results Hot - started and two - step PCR can improve the efficiency and decrease non - specific amplification of ApoE gene PCR. The allele frequency of ApoE4 is increasing in Senile osteoporosis group （ P 〈 0.05 ）. Conclusion Hot - started and two - step PCR can improve the efficiency and specificity of ApoE gene amplification. It will play an important role in the process of searching ApoE gene type, and ApoE4 allele maybe have close relationship with Senile osteoporosis.