Matsubara K, Koide Y, Kobayashi A, Kaida R, Takeda S, Matsuda E, Matsuoka Y, Yoshida T O
Clinical Science Center, Diagnostic Technology Labs of Chugai Pharmaceutical Company, Tokyo, Japan.
Hum Immunol. 1992 Oct;35(2):132-9. doi: 10.1016/0198-8859(92)90021-e.
A rapid and accurate detection of HLA class II antigen is essential for transplantation and for the understanding of disease susceptibility. Recent molecular genetic studies have revealed that the number of class II loci and the number of alleles at these loci are greater than had been previously detected. It is, therefore, of great importance to detect these extensive polymorphisms. A great deal of effort has been made on identification of individual HLA class II specificities at the DNA level, called "DNA typing." What seems to be lacking, however, is handling simplicity. Here we accomplished a simple method for HLA-DRB1 typing based on hybridization of acridinium-ester (AE)-labeled DNA probes to amplified DNA. This method is called hybridization protection assay (HPA). By using 13 AE-labeled probes, 20 homozygous B-cell lines and leukocytes from 80 healthy individuals were typed by HPA. The results were completely consistent with those obtained by polymerase chain reaction--restriction fragment length polymorphism. This method is suitable for mass screening because of its procedural simplicity and swiftness.
快速准确地检测人类白细胞抗原（HLA）II类抗原对于移植以及理解疾病易感性至关重要。最近的分子遗传学研究表明，II类基因座的数量以及这些基因座上等位基因的数量比以前检测到的要多。因此，检测这些广泛的多态性非常重要。在DNA水平上鉴定个体HLA II类特异性（即“DNA分型”）已经付出了巨大努力。然而，似乎缺乏操作简便性。在此，我们基于吖啶酯（AE）标记的DNA探针与扩增DNA的杂交，完成了一种用于HLA-DRB1分型的简单方法。这种方法称为杂交保护分析（HPA）。通过使用13种AE标记的探针，采用HPA对20个纯合B细胞系和80名健康个体的白细胞进行了分型。结果与通过聚合酶链反应-限制性片段长度多态性获得的结果完全一致。由于其操作简便快捷，该方法适用于大规模筛查。
