HLA-DQB1-PCR-SSP方法的建立及其在骨髓移植中的应用研究

为了改进骨髓移植中供、受体配型技术,在已有技术的基础上改进、建立了新的HLA-DQB1-PCR-SSP方法。利用澳大利亚皇家医院赠送的HLA标准分型细胞对自行设计引物的特异性进行鉴定,发现该引物设计合理、扩增特异。通过内部阳性对照引物的设立,可较容易地将纯合子及杂合子分型细胞分开,并对个体DNA进行了尝试。结论表明,HLA-DQB1-PCR-SSP快速、简便、灵敏、重复性好,结果易于判断,适于小样本及急症病人的器官及骨髓移植配型,是比较理想的HLA-Ⅱ类基因分型方法。

Establishment of HLA-DQBI-PCR-SSP method and its application in bone marrow transplantation

It has been established that mismatches at certain important loci of the human leucocyte antigen (HLA) in donor/recipient pairs assignment increase the incidence of GVHD or the risk of graft failure following organ transplantation. Thus, the correct identification of HLA allele is the key step to successful transplantation. In order to improve the donor and recipient matching methods in bone marrow transplantation, we improved and established a new HLA-DQB1-PCR-SSP method by basing on already established techniques. Special attention was directed towards the simple technical aspects of procedures , the level of typing resolution and the speed of data analysis. By using DQB1 standard typing cells which are gifts from Royal Hospital of Australia, we verified the specificity of the primers which were designed by ourselves . The result showed that the designed primers were specific and reasonable. By adding internal positive controls , we can easily distinguish homozygous or heterozygous cells, as well as homozygous or heterozygous individuals. Our findings suggested that HLA-DQB1-PCR-SSP method is a fast, sample, sensitive and reproducible method, its results can be easily interpreted, and it may be suitable for small numbers of samples, urgent cases of organ or bone marrow transplant donor and recipient typings.