Forssmann U, Mytilineos J, Scherer S, Opelz G
Department of Transplantation Immunology, Institute of Immunology, University of Heidelberg, Germany.
Transpl Int. 1994;7 Suppl 1:S515-8. doi: 10.1111/j.1432-2277.1994.tb01432.x.
There is preliminary evidence that matching for HLA-DQ is important for kidney graft survival. We developed a method for HLA-DQA typing based on the PCR-SSP principle. The procedure consisted of three steps: DNA isolation, PCR amplification and visualization of the PCR product under UV light. For the identification of all currently known DQA1 alleles, we designed 18 different primers that allowed typing for the specificities DQA1*0101, *1012, *0103, *0104, *0201, *03, *0401, 0501 and DQA10601. For the typing of a single individual, 12 PCR mixes were needed, each containing a primer pair specific for a certain allele group, and a pair of control primers that amplified a non-polymorphic region. The time required for this procedure was approximately 3 h from the time of blood collection. Comparison of this method with DQA typing by the RFLP method in 151 individuals revealed only a single discrepancy. The method can be easily applied for prospective cadaver donor typing.
有初步证据表明，HLA-DQ配型对肾移植存活很重要。我们基于PCR-SSP原理开发了一种HLA-DQA分型方法。该程序包括三个步骤：DNA分离、PCR扩增以及在紫外光下观察PCR产物。为鉴定所有目前已知的DQA1等位基因，我们设计了18种不同引物，可用于DQA1*0101、*1012、*0103、*0104、*0201、*03、*0401、0501和DQA10601特异性分型。对于单个个体的分型，需要12种PCR混合物，每种混合物包含一对针对特定等位基因组的引物对，以及一对扩增非多态性区域的对照引物。从采血开始，该程序所需时间约为3小时。在151名个体中，将该方法与RFLP法进行DQA分型比较，仅发现一处差异。该方法可轻松应用于前瞻性尸体供体分型。
