Development of a new HLA-DNA typing method applicable in the nation-wide renal transplantation network.

器官移植供体和受体的HLA分型对于非亲属尸体器官移植的公平和效率具有重要意义。我们已经开发了一种新的HLA-DNA分型方法，使用靶向基因组DNA的RNA扩增方法，转录介导扩增（TMA）和基于化学发光的杂交保护试验（HPA），这是一种序列特异性寡核苷酸（SSO）方法。在本研究中，我们将聚合酶链反应（PCR）与HPA法相结合，进一步分析DR6抗原群的精细等位基因。用dr356特异性引物PCR扩增121份DNA样本，用HPA法检测。DRB1^<** >3等位基因组确定分型的等位基因有10个（DRB1^<**>130l-1307, ^<**>1310-1312）， DRB1^<** >4等位基因组确定分型的等位基因有11个（DRB1^<**> l401-1408, ^<**>1411, **>1412, ^<**>l417）。然而，某些等位基因组合（^<**>1301/02,^<**>1140l/07）没有明确分型，需要额外的引物组合。PCR-HPA法的操作时间为DNA提取1小时，扩增2小时，检测1小时，共4小时。因此，PCR-HPA法是一种简单、快速、无同位素、准确的HLA-DNA分型方法，将保证器官移植的公平和效率。

HLA typing of both donor and recipients in organ transplantation is important from the stand point of equity and efficiency in unrelated cadaver organ transplantation. We have already developed a new HLA-DNA typing method using a RNA amplification method targeting at the genomic DNA,Transcription-Mediated Amplification (TMA) and a chemiluminescence-based Hybridization Protection Assay (HPA), which is one type of sequence-specific oligonucleotide (SSO) methods. In this research, we combined polymerase chain reaction (PCR) with HPA method, to analyze further fine alleles of the DR6 antigen group. One-hundred and twenty-one DNA samples were amplified with DR356-specific primers by PCR and detected by HPA method. Among DRB1^<**>13 allele group, 10 alleles (DRB1^<**>130l-1307, ^<**>1310-1312) were derfinitely typed, and among DRB1^<**>14,11 a11eles (DRBl^<**>l401-1408, ^<**>1411^<, **>1412, ^<**>l417). However, a certain combination of alleles (^<**>1301/02, ^<**>1140l/07) were not typed unequivocally and need additional primer combinations. The procedure time required for PCR-HPA method is one hour for extraction of DNA,2 hours for amplification, and one hour for detection, a total of 4 hours. Thus, PCR-HPA method for HLA-DNA typing is a simple, fast, non-isotopic and accurate method, which will guarantee the equity and efficiency in organ transplantation.