通过限制性片段长度多态性分析对粒细胞特异性NA抗原进行分型。

Typing of the granulocyte-specific NA antigens by restriction fragment length polymorphism analysis.

作者信息

Stein E L, Bux J, Santoso S, Mueller-Eckhardt C

机构信息

Institute for Clinical Immunology and Transfusion Medicine, University of Giessen, Germany.

出版信息

Br J Haematol. 1994 Jun;87(2):428-30. doi: 10.1111/j.1365-2141.1994.tb04939.x.

DOI:10.1111/j.1365-2141.1994.tb04939.x

PMID:7947295

Abstract

An RNA-based method has been developed to genotype donors for the granulocyte-specific alloantigens NA1 and NA2. mRNA was isolated from granulocytes, reversely transcribed into cDNA and amplified using an Fc-gamma-receptor III-1 sequence-specific primer in the polymerase chain reaction (PCR). PCR products were analysed by restriction fragment length polymorphism (RFLP) using the restriction endonuclease Taq I, which provided a distinct restriction fragment pattern corresponding to the NA alleles. 17 donors were typed by PCR-RFLP and the results were in close accordance with those obtained by serological phenotyping by granulocyte immunofluorescence and the antigen capture assay MAIGA.

摘要

已开发出一种基于RNA的方法，用于对粒细胞特异性同种异体抗原NA1和NA2的供体进行基因分型。从粒细胞中分离出mRNA，逆转录成cDNA，并在聚合酶链反应（PCR）中使用Fc-γ受体III-1序列特异性引物进行扩增。使用限制性内切酶Taq I通过限制性片段长度多态性（RFLP）分析PCR产物，该酶提供了与NA等位基因相对应的独特限制性片段模式。通过PCR-RFLP对17名供体进行基因分型，结果与通过粒细胞免疫荧光血清学表型分析和抗原捕获测定MAIGA获得的结果高度一致。