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通过DNA亚类特异性扩增和等位基因特异性探针在基因组水平上测定同种异型G1m(f)和G1m(z)

Determination of allotypes G1m(f) and G1m(z) at the genomic level by subclass-specific amplification of DNA and use of allele-specific probes.

作者信息

Balbín M, Grubb A, Abrahamson M, Grubb R

机构信息

Department of Clinical Chemistry, University of Lund, Sweden.

出版信息

Exp Clin Immunogenet. 1991;8(2):88-95.

PMID:1789994
Abstract

Two oligonucleotide primers were used for selective enzymatic amplification of a DNA segment encoding a major portion of the first constant region domain (CH1) of the human IgG1 heavy chain. The selective amplification was confirmed by use of subclass-specific oligonucleotide probes. Two 15-mer oligonucleotides, hybridizing with the alleles for the allotypes G1m(f) and (z), respectively, could then be used for determination at the genomic level of these two truly allelic allotypes. Serum and DNA samples from 12 individuals, one of them with a considerable amount of anti-Gm(f) antibodies, were used for allotype assignment by classical serological methods and by the new method operating at the genomic level. The resulting classifications agreed completely, demonstrating the reliability of the new method.

摘要

使用两种寡核苷酸引物对编码人IgG1重链第一恒定区结构域(CH1)主要部分的DNA片段进行选择性酶促扩增。通过使用亚类特异性寡核苷酸探针证实了选择性扩增。然后,分别与同种异型G1m(f)和(z)的等位基因杂交的两种15聚体寡核苷酸可用于在基因组水平上确定这两种真正的等位基因同种异型。来自12个人的血清和DNA样本(其中一人含有大量抗Gm(f)抗体)用于通过经典血清学方法和在基因组水平上操作的新方法进行同种异型分型。所得分类完全一致,证明了新方法的可靠性。

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