直接半巢式PCR结合测序鉴定ApoE基因型的方法的建立

Establishment of Method for Identifying ApoE Genotype by Direct Semi-Nested PCR Combined with Sequencing

摘要: 目的：建立一种直接使用口腔上皮细胞为模板PCR扩增ApoE基因rs429358和rs7412位点从而采用桑格法测序鉴定ApoE基因型的方法。方法：自行设计3条引物，以口腔上皮细胞粗处理物为模板，通过半巢式PCR扩增包含ApoE基因2个SNP位点的靶片段，PCR产物经桑格法测序鉴定样本基因型。结果：所检样本能扩增出预期大小的PCR产物，测序峰图清晰。结论：成功建立了一种直接半巢式PCR结合测序鉴定ApoE基因型的方法，有良好的应用前景。

Abstract: Objective: To establish a method for identifying ApoE genotype by PCR and Sanger sequencing directly using oral epithelial cells. Meth-ods: Self-designed three primers were used to amplify the target fragments containing two SNP loci of ApoE gene by semi-nested PCR. The PCR products were sequenced to identify the genotype of the samples. Results: The PCR products of the expected size can be amplified from the tested samples, and the sequencing peak diagram is clear. Conclusion: A method of direct semi-nested PCR com-bined with sequencing to identify ApoE gene was successfully established, which has a certain ap-plication value.