Corell A, Pay A L, Little A M, Hoddinott M A, Argüello J R, Borton M, Dunne C, Ogilvie H, O'Shea J, Madrigal J A, Marsh S G
Anthony Nolan Research Institute, The Royal Free Hospital, London, United Kingdom.
Tissue Antigens. 2000 Jul;56(1):82-6. doi: 10.1034/j.1399-0039.2000.560111.x.
We show here the use of reference strand mediated conformation analysis (RSCA) to unambiguously resolve the HLA-DRB1 typing of two individuals which were selected as potential unrelated donors for bone marrow transplantation (BMT). In the first case, both sequence-specific primer (SSP) amplification and sequence-specific oligonucleotide probing (SSO), routinely used in different tissue typing laboratories gave, for the two unrelated donors, the same ambiguous typing of HLA-DRB104011+0403 or DRB10407+0413. In this case sequence-based typing (SBT) was not the method of choice to resolve the situation, due to the sequence ambiguities of these two given combinations. RSCA of both samples, using homozygous typing cells (HTCs) for DRB104011, 0403 and 0407 as internal controls, gave the unambiguous result that both donors were HLA-DRB104011+0403. In the second case, a donor was typed as DRB11102+1103 by SSP, while SSO excluded the DRB11102 allele. The patient was unambiguously typed as DRB11101+1103 by both techniques. RSCA, using DNA from reference cell lines as internal controls, gave the unambiguous typing that the donor was DRB1*1103 homozygous.
我们在此展示了参考链介导构象分析（RSCA）的应用，以明确解析两名个体的HLA - DRB1分型，这两名个体被选为骨髓移植（BMT）的潜在非亲属供体。在第一个案例中，不同组织分型实验室常规使用的序列特异性引物（SSP）扩增和序列特异性寡核苷酸探针杂交（SSO），对于这两名非亲属供体，给出了相同的模糊HLA - DRB1分型结果，即DRB104011 + 0403或DRB10407 + 0413。在这种情况下，基于序列的分型（SBT）并非解决该问题的首选方法，因为这两种给定组合存在序列模糊性。使用DRB104011、0403和0407的纯合分型细胞（HTC）作为内部对照，对两个样本进行RSCA，得出明确结果，即两名供体均为HLA - DRB104011 + 0403。在第二个案例中，一名供体通过SSP分型为DRB11102 + 1103，而SSO排除了DRB11102等位基因。两种技术均明确将患者分型为DRB11101 + 1103。使用来自参考细胞系的DNA作为内部对照进行RSCA，得出明确分型结果，即供体为DRB1*1103纯合子。
