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使用等位基因特异性或组特异性扩增进行II类HLA抗原的DNA分型。I. HLA-DR4亚组的分型

DNA typing for class II HLA antigens with allele-specific or group-specific amplification. I. Typing for subsets of HLA-DR4.

作者信息

Gao X J, Fernandez-Vina M, Shumway W, Stastny P

机构信息

Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas 75235.

出版信息

Hum Immunol. 1990 Jan;27(1):40-50. doi: 10.1016/0198-8859(90)90094-6.

Abstract

DNA sequences that distinguish the subsets of HLA-DR4 are also found on several other alleles. This makes typing of heterozygotes with oligonucleotide probes quite impractical. We have therefore developed a procedure in which, in a first step, DNA of the genes to be analyzed is amplified selectively, using group-specific primers. In the case of DR4-DRB1, a primer matching codons 5 to 13 when used in the polymerase chain reaction resulted in products that were entirely suitable for typing for the DR4 subsets. Using appropriate probes, eight distinct subsets were identified. However, only six of them were represented in a normal North American Caucasian panel. In conjunction with a method for rapid DNA extraction, the procedure offers a simple, highly specific and reproducible method for determining subtypes of HLA-DR4 that at present cannot be recognized by serologic methods.

摘要

在其他几个等位基因上也发现了区分HLA - DR4亚群的DNA序列。这使得用寡核苷酸探针鉴定杂合子变得非常不切实际。因此,我们开发了一种方法,第一步,使用组特异性引物选择性扩增待分析基因的DNA。对于DR4 - DRB1,在聚合酶链反应中使用与第5至13密码子匹配的引物,产生的产物完全适合用于DR4亚群的分型。使用适当的探针,鉴定出八个不同的亚群。然而,在正常的北美白种人样本中只出现了其中六个。结合快速DNA提取方法,该程序提供了一种简单、高度特异且可重复的方法来确定目前血清学方法无法识别的HLA - DR4亚型。

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