通过巢式聚合酶链反应-序列特异性引物对HLA-B5交叉反应组进行低分辨率DNA分型。

Low resolution DNA typing of the HLA-B5 cross-reactive group by nested PCR-SSP.

作者信息

Hein J, Böttcher K, Grundmann R, Kirchner H, Bein G

机构信息

Institute of Immunology and Transfusion Medicine, University of Lübeck Medical School, Germany.

出版信息

Tissue Antigens. 1995 Jan;45(1):27-35. doi: 10.1111/j.1399-0039.1995.tb02411.x.

DOI:10.1111/j.1399-0039.1995.tb02411.x

PMID:7725308

Abstract

We have established a DNA typing system for the HLA-B5 serologically cross-reactive group (CREG) by means of a two-step PCR amplification with nested sequence-specific primers (nPCR-SSP). The present study provides a low resolution definition of the HLA-B5 CREG, i.e. identifying polymorphism equivalent to serology. Two different primer combinations allow group-specific amplification of all HLA-B5 CREG alleles and other related HLA class I alleles from genomic DNA. The amplified DNA is subjected to a second amplification step using eleven nested primer pairs. This assay permits the detection of the HLA-B5 CREG specificities B35, B51, B52, B53, and B7801 in all homozygous and heterozygous combinations. Sensitivity and specificity as judged by a blind quality control study investigating a reference panel (n = 50) is 100%. Extension of this approach should allow rapid DNA typing of all serologically defined HLA-B specificities by nPCR-SSP.

摘要

我们通过使用巢式序列特异性引物的两步聚合酶链反应（nPCR - SSP）建立了一种针对HLA - B5血清学交叉反应组（CREG）的DNA分型系统。本研究提供了HLA - B5 CREG的低分辨率定义，即识别与血清学等效的多态性。两种不同的引物组合可从基因组DNA中对所有HLA - B5 CREG等位基因和其他相关的HLA I类等位基因进行组特异性扩增。扩增的DNA使用11对巢式引物对进行第二步扩增。该检测方法能够检测所有纯合和杂合组合中的HLA - B5 CREG特异性B35、B51、B52、B53和B7801。通过对一个参考样本组（n = 50）进行的盲法质量控制研究判断，其灵敏度和特异性均为100%。扩展这种方法应能通过nPCR - SSP对所有血清学定义的HLA - B特异性进行快速DNA分型。