定量PCR检测HLA-C等位基因表达方法的建立和初步应用

发布时间：2018-05-07 01:39

  本文选题：定量PCR + HLA-C　； 参考：《中国免疫学杂志》2016年08期

【摘要】：目的:建立实时定量PCR方法检测同一个体两条不同的HLA-C等位基因的表达水平。方法:构建HLAⅠ类等位基因外显子2和3数据库,设计等位基因特异性系列引物,建立等位基因定量PCR方法;在数据库和835例汉族人外周血标本中对方法进行理论和实验验证;检测并比较20对肝癌和癌旁组织标本中HLA-C等位基因的表达水平。结果:本实验设计的20对引物能检测23种杂合基因型中不同的HLA-C等位基因,覆盖汉族人群中出现频率大于0.96%的个体;55%(11/20)的肝癌组织中HLA-C位点两条等位基因表达水平变化不一致。结论:成功建立了针对汉族人群的HLA-C等位基因表达水平的检测方法;肝癌组织中普遍存在着两条HLA-C等位基因表达水平变化不一致的现象。
[Abstract]:Objective: to establish a real-time quantitative PCR method to detect the expression of two different HLA-C alleles in the same body. Methods: HLA class I allele exons 2 and 3 databases were constructed, allele specific series primers were designed and allele quantitative PCR method was established, and the method was verified theoretically and experimentally in the database and 835 peripheral blood samples of Han nationality. The expression level of HLA-C allele in 20 pairs of HCC and adjacent tissues was detected and compared. Results: the 20 pairs of primers designed in this experiment can detect different HLA-C alleles in 23 heterozygous genotypes, and the two alleles of HLA-C locus in HCC tissues with frequency of more than 0.96% and 0.96% were not consistent with each other. Conclusion: a method of detecting HLA-C allele expression in Han population was successfully established and two HLA-C alleles were found to be inconsistent in HCC tissues.
【作者单位】： 东南大学医学院;南京医科大学第一附属医院普外科肝脏外科;
【基金】：国家自然基金青年科学基金项目(81201601)资助
【分类号】：R440
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本文编号：1854855