Smrzka O W, Faé I, Pickl W F, Fischer G F
Institute for Blood Group Serology University of Vienna, Austria.
Tissue Antigens. 1991 May;37(5):205-10. doi: 10.1111/j.1399-0039.1991.tb01873.x.
Locus HLA-DRB3 codes for the serologically defined supertypic specificity DRw52 in HLA-DR3, -5 and -w6 haplotypes. Three specificities of DRw52 (DRw52a, -b and -c) can further be distinguished by cellular techniques or by DNA typing with allele-specific oligonucleotide probes. These specificities were recently reported to have significant importance in antigen presentation. To avoid a time-consuming hybridization procedure, we have developed a simple typing system using PCR and subsequent digestion by allele-specific restriction endonucleases. A system was established with locus-specific amplification of HLA-DRB3 and digestion by the enzymes KpnI, ScaI and HinfI which recognize unique restriction sites within the amplified region. This allowed HLA-DRB3 typing on agarose gel by determining whether the amplification product has been digested or not. This typing system was compared to conventional oligotyping by analyzing 145 RFLP-typed individuals for their DRw52 specificity using both methods. Agarose typing correlated well with oligotyping and was shown to be more simple and practical even in heterozygous individuals.
HLA - DRB3基因座编码HLA - DR3、- 5和 - w6单倍型中血清学定义的超型特异性DRw52。DRw52的三种特异性（DRw52a、- b和 - c）可以通过细胞技术或使用等位基因特异性寡核苷酸探针的DNA分型进一步区分。最近有报道称这些特异性在抗原呈递中具有重要意义。为避免耗时的杂交程序，我们开发了一种使用聚合酶链反应（PCR）并随后用等位基因特异性限制性内切酶消化的简单分型系统。建立了一个系统，对HLA - DRB3进行基因座特异性扩增，并用识别扩增区域内独特限制性位点的KpnI、ScaI和HinfI酶进行消化。通过确定扩增产物是否被消化，可在琼脂糖凝胶上进行HLA - DRB3分型。通过使用这两种方法分析145个经限制性片段长度多态性（RFLP）分型的个体的DRw52特异性，将该分型系统与传统的寡核苷酸分型进行了比较。琼脂糖分型与寡核苷酸分型相关性良好，并且即使在杂合个体中也显示出更简单和实用。
