用PCR单链构象多态性快速判别个体间HLA－DP相容性并检出新等位基因

先确定和比较ＨＬＡ纯合细胞ＤＰＡＩ和ＤＰＢＩ基因ＰＣＲ扩增产物的单链构象多态性（ＰＣＲ－ＳＳＣＰ），然后分析了２０对无血缘关系个体单链ＤＮＡ及其两两混合状态下的构象多态性变化，在分子水平测定异体间ＤＰ匹配性，并和已有的ＤＰ基因分型（ＰＣＲ－ＲＦＬＰ）格局相比较，结果表明，ＲＦＬＰ不同的个体其ＳＳＣＰ必定不同；而ＲＦＬＰ相同的个体，ＳＳＣＰ一般相同，但有例外。从例外个体异常的ＳＳＣＰ格局中发现了新ＤＰ等位基因。本研究证实ＰＣＲ－ＳＳＣＰ在检测等位基因之间微小差异方面十分敏感，并具有简单、快速、经济等优点，对移植中的ＤＰ交叉配型有实际应用价值。

SINGLE STRAND CONFORMATION POLYMORPHISM OF PCR-AMPLIFIED DNA,AN APPROACH FOR CROSS-MATCHING TESTOF HLA-DP GENES BETWEEN UNRELATED SUBJECTS AND FOKDETECTION OF NOVEL DP ALLELES

With genotyping results of allele specificities by restriction fragment Iength polymorphismof PCR-amplified HLA-DPA1 and DPB1 genes ( PCR-RFLP),different patterns of singlestrand conformation polymorphism of the amplified gene products (PCR-SSCP)were comparedin 6 Chinese, and 4 reference HLA homologous cell of International HistocompabilityWorkshops. And 20 groups of unrelated individuals were further subjected as tentative donor-re-cipient pairs to cross -matching test by PCR-SSCP,which was recently adopted as an approachfor donor-recipient screening at DNA level in transplantation. Our results indicate that differentDP alleles typed by PCR-RFLP between donors and recipients always gave distinct PCR-SSCPpatterns; and the subjects,with identical PCR-RFLP-typing results gave well-matched SSCP ex-cept three pairs,in which two tentative recipients were identified to have a novel DPB1 or DPA1allele by peculiar SSCP patterns.These observations implicate that with advantage such as sensi-tivity, simplicity and rapidity,the PCR-SSCP method can be used both in seeking DP well-matched donor-recipient pairs in population even they have not been HLA genotyped,and in i-dentification of novel DP alleles preliminarily.