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目的 将PCR指纹图用于个体间HLA DP的基因水平交叉配型 ,确定供 受体间DPB基因的相容性。方法 引入猴的类DPB等位基因的扩增产物作通用性异源双链生成物 (UHG) ,使原不能产生PCR指纹图的DPB基因扩增产物在非变性PAGE中显示特定的“卫星条带” ,从而可用于供 受体间HLA DP基因的相容性测定。结果 11株HLA纯合的B淋巴细胞系和随机个体进行PCR DPB指纹图及交叉配型分析 ,能敏感地反映个体间DP基因的差异 ,特别是在纯合子DPB 0 2 0 1和杂合子DPB 0 40 2 0 5 0 1中初步发现了可能存在着未能被PCR RFLP区分的新的亚型或新的等位基因 ,但是尚不能区分DPB 0 2 0 1和DPB 0 40 2两个等位基因 ,有待今后进一步探索。结论 初步表明该技术可有效判别供 受体DPB基因匹配性 ,并有简单、快速等优点。
OBJECTIVE: To use PCR fingerprinting for HLA DP cross-genotyping between individuals to determine the compatibility of DPB genes for receptors. Methods The amplification product of monkey DPB allele was introduced into universal heteroduplex product (UHG). The amplification products of DPB gene that did not produce PCR fingerprinting were displayed as “ Band, ”thus allowing for the determination of the compatibility of the HLA DP genes between the receptors. Results Eleven HLA homozygous B lymphocyte lines and random individuals were analyzed by PCR DPB fingerprinting and crossmatch analysis, which could sensitively reflect the differences of DP genes among individuals, especially in homozygous DPB 0 2 0 1 and heterozygous DPB A new subtype or new allele that could not be distinguished by PCR RFLP was initially found in 0 40 2 0 5 0 1 but no distinction was made between the two alleles of DPB 0 2 0 1 and DPB 0 40 2 Gene, to be further explored in the future. Conclusions Preliminary results show that the technology can effectively discriminate donor-donor DPB gene matching, and has the advantages of simple and rapid.