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直接巢式PCR结合测序检测5-HT1A基因C-1019G多态性的方法的建立
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Abstract:
目的：5-HT1A基因C-1019G多态性与多种神经精神疾病如抑郁症、精神分裂症、焦虑症等的发生风险、症状严重程度及治疗效果存在关联。本研究旨在建立一种直接巢式PCR结合测序检测该位点的方法。方法：以口腔上皮细胞粗处理物为材料，通过直接巢式PCR扩增包含5-HT1A基因C-1019G位点的靶片段，PCR产物经桑格测序鉴定基因型。结果：所检样本均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巢式PCR结合测序鉴定5-HT1A基因型的方法，有良好的应用前景。
Objective: The C-1019G polymorphism of the 5-HT1A gene has been associated with the risk of occurrence, symptom severity, and treatment outcome of a variety of neuropsychiatric disorders, such as depression, schizophrenia, and anxiety disorders. The aim of this study was to establish a direct nested PCR combined with sequencing to detect this locus. Methods: The target fragment containing the C-1019G locus of the 5-HT1A gene was amplified by direct nested PCR using crude processed oral epithelial cells, and the PCR product was genotyped by Sanger sequencing. Results: PCR products of expected size were amplified from all the samples tested, and the sequencing peaks were clear. Conclusion: A direct nested PCR combined with sequencing method was successfully established to identify the genotypes of 5-HT1A gene, which has good prospects for application.
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