人类白细胞抗原基因分型与微珠反应格局  

Human leucocyte antigen genotyping and beads reaction patterns

作　　者：王大明[1] 何柳媚[1] 张红[1] 魏天莉[1] 李茜[1] 邹红岩[1] 

机构地区：[1]深圳市血液中心,广东省深圳市518035

出　　处：《中国组织工程研究与临床康复》2010年第53期9960-9964,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：背景:Flow-rSSO结合流式细胞分析技术和PCR-SSO技术的高通量分型方法,是目前国外人类白细胞抗原分型中应用最多的方法。目的:筛选同组不同等位基因之间的微珠反应格局,加强人类白细胞抗原基因分型方法的标准化,使分型向最快最准确的方向发展。方法:使用TECAN DNA全自动工作站从457份中华骨髓库捐献者全血样本中提取基因组DNA,DNA样本用One Lambda rSSO HLA-A、B和DRB1基因分型高分试剂盒进行PCR扩增、分子杂交和Luminex流式磁珠分析仪检测。参考One Lambda公司提供等位基因微珠格局表(HLA-A,Band DRB1)分析常见等位基因,并对常见等位基因与微珠的相关性进行研究。结果与结论:同一组常见等位基因除相同微珠反应格局外,与某个或某几个有区别的微珠是密切相关的。就临床组织器官移植而言,采用中分辨度DNA分型技术是最佳选择,一方面有利于快速筛选,另一方面能够降低匹配难度。就科研工作而言,一般采用高分辨率分型方法。BACKGROUND：Flow-rSSO in combination with flow cytometry and PCR-SSO with high throughout subtyping are the most commonly used methods for human leucocyte antigen（HLA） genotyping at present in the countries outside China.OBJECTIVE：To select the beads reaction patterns of different alleles in the same group and standardize the HLA genotype method,hopefully to require a more specific and rapid genotype development.METHODS：Genomic DNA was extracted automatically from 457 blood samples using TECAN DNA workstation.Each DNA sample was subjected to PCR amplification,molecular hybridization and Luminex magnetic beads analysis using One lambada rSSO HLA-A,B and DRB1 commercial kit.With reference to the HLA-A,B and DRBI provided by One Lambda Inc.,the common alleles were analyzed and the correlation between common alleles and beads was studied.RESULTS AND CONCLUSION：The common alleles in the same group are closely related to a certain or some different beads except the same beads reaction situation.To the organ transplantation in the clinic,the adoption of DNA genotype of middle resolution is the best choice.It not only contributes to quick screening,but also reduces the difficulty of matching.The high resolution genotype is commonly adopted in research work.

关 键 词：基因组DNA 全血 中华骨髓库 HLA基因分型 

分 类 号：R318[医药卫生—生物医学工程]