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

开发适用于全国肾移植网络的新 HLA-DNA 分型方法。

基本信息

批准号：
08457313
负责人：
MAEDA Hiroo
金额：
$ 4.93万
依托单位：
Saitama Medical School
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08457313/
关键词：
Kidney transplantation HLA DNA typing HPA

项目摘要

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.

从非血缘关系尸体器官移植的公平性和效率的角度来看，器官移植中供受者的HLA分型是重要的。我们已经开发了一种新的HLA-DNA分型方法，该方法使用靶向基因组DNA的RNA扩增方法、转录介导扩增（TMA）和基于荧光的杂交保护测定（HPA），这是一种序列特异性寡核苷酸（SSO）方法。本研究将聚合酶链反应（PCR）与HPA法相结合，进一步分析DR 6抗原组的精细等位基因。采用PCR方法对121份DNA样本进行扩增，并用HPA法进行检测。在DRB_（1 *）13个等位基因组中，有10个等位基因（DRB_（1 *）1301 -1307，DRB_（1 *）1310-1312），在DRB_（1 *）14个等位基因组中，有11个等位基因（DRB_（1 *）1401 -1408，DRB_（1 **）1411，DRB_（1 **）1412，DRB_（1 *）1417）。然而，等位基因的特定组合（^<**>1301/02，^<**> 11401/07）不能明确分型，需要额外的引物组合。PCR-HPA方法所需的程序时间为DNA提取1小时，扩增2小时，检测1小时，共4小时。因此，PCR-HPA法是一种简便、快速、非同位素、准确的HLA-DNA分型方法，可保证器官移植的公平性和效率。