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目的:探索应用焦磷酸微测序技术进行HLADRB基因型分析.方法:PCR扩增外显子2基因片段后,应用焦磷酸微测序技术进行实时测序和HLADRB基因分型.结果:磷酸微测序反应可以判读HLADRB基因的多态性位点,最长可判读90个核苷酸,采用特异性等位基因和PCR反应所得到的纯化DNA对血样中的杂合子进行分析,测序结果与HLA数据库基因序列比较,可准确进行HLADRB基因型分析.结论:用焦磷酸微测序技术进行HLADRB基因型分析具有高分辨率的优点,该方法可应用于临床器官移植的供体/受体筛查.
OBJECTIVE: To explore the genotype analysis of HLADRB using pyrosequencing technology.Methods: Real-time sequencing and HLADRB genotyping were performed using pyrosequencing technology after amplification of exon 2 gene fragment.Results: Phosphate micro sequencing reaction could Interpretation of HLADRB gene polymorphic sites, the maximum interpretation of 90 nucleotides, using specific alleles and PCR reaction of purified DNA samples of blood heterozygous analysis, sequencing and HLA database gene sequence Compared with HLADRB genotypes.Conclusion: HLADRB genotype analysis using pyrosequencing with high resolution has the advantages of high resolution, the method can be applied to clinical donor / donor screening.