Blasczyk R, Mohr M, Zimmermann R, Schwella N, Huhn D
Blutbank, Abteilung für Innere Medizin mit Schwerpunkt Hämatologie und Onkologie, Universitätsklinikum Rudolf Virchow, Freie Universität, Berlin.
Infusionsther Transfusionsmed. 1994 Dec;21(6):401-4. doi: 10.1159/000223019.
The allelic diversity of HLA-DPB1 antigens can be determined at the DNA level after PCR amplification. The pattern of polymorphism at the DPB1 locus makes it difficult to unambiguously assign all genotypes in a typing system using one single pair of generic primers.
We apply here a simple technique based on the reverse dot blot analysis to the typing of HLA-DPB1 alleles. In order to increase its resolution, a group-specific amplification based on sequence variations of the polymorphic region F was used subdividing the HLA-DPB1 alleles in 2 nonoverlapping families. A separate analysis was then performed within each group of alleles.
Using these 2 primer pairs, 21 group 1 and 30 group 2 alleles were separately amplified. From 1,378 possible allele combinations for DPB1*0101-5301 only 33 gave ambiguous typing results compared to 61 using a single pair of generic primers.
This procedure provides a rapid and simple HLA-DPB1 genotyping. Especially in heterozygotes the hybridization patterns were easier to interpret. The utilization of group-specific amplification substantially reduced ambiguous typing results.
聚合酶链反应（PCR）扩增后，可在DNA水平确定HLA - DPB1抗原的等位基因多样性。DPB1基因座的多态性模式使得在使用一对通用引物的分型系统中难以明确确定所有基因型。
我们在此将基于反向斑点杂交分析的一种简单技术应用于HLA - DPB1等位基因分型。为了提高其分辨率，基于多态性区域F的序列变异进行分组特异性扩增，将HLA - DPB1等位基因分为2个不重叠的家族。然后在每个等位基因组内进行单独分析。
使用这2对引物，分别扩增出21个第1组和30个第2组等位基因。对于DPB1*0101 - 5301的1378种可能的等位基因组合，与使用一对通用引物时的61种相比，只有33种产生了模糊的分型结果。
该方法提供了一种快速简便的HLA - DPB1基因分型方法。特别是在杂合子中，杂交模式更易于解释。分组特异性扩增的应用大大减少了模糊的分型结果。
