A novel real-time quantitative PCR method for analyzing HLA-C allele expression level / 中国免疫学杂志

ABSTRACT

Objective: To establish real-time qPCR method to analyze each HLA-C allele expression level of individual.Methods:Database including exon 2&3 sequences of HLA class Ⅰalleles was built ,HLA-C allelic specific primers were designed and the real-time quantitative PCR method for analyzing HLA-C allele expression level was built .The allelic specificity of these primers were confirmed in database and 835 normal peripheral blood samples of Han population .The mRNA level of each HLA-C allele from 20 pairs of liver tumor tissues and non-tumor tissues was analyzed by the qPCR method we built .Results:20 pairs of allelic specific primers were designed to distinguish the two HLA-C alleles of each individual with frequency over 0.96%in 835 cases of Han population.Among 55%of the liver cancer tissues ,the expression levels of the two HLA-C alleles from the same tumor tissue changes differently compared to that of the relevant non-tumor tissue.Conclusion:This study provides method for HLA-C allele expression level analysis of Han population and each HLA-C allele expression level is inconsistent of liver cancer tissue .