HLA-DR generic typing by AFLP.
HLA-DR generic typing by AFLP.
复制标题
AFLP 的 HLA-DR 通用分型。
DOI：
10.1111/j.1399-0039.1991.tb01884.x
复制
发表时间：
1991
期刊：
影响因子：
--
通讯作者：
Yunis,EJ
中科院分区：
文献类型：
--
作者：
Yunis,I;Salazar,M;Yunis,EJ
我们已经描述了一种实用且廉价的方法，由此可以对任何个体进行分型并将其分配给14种通用HLA-DR类型中的任何一种：DR 1、DRwl 5、DRwl 6、DRwl 7、DRwl 8、DR 4、DRwll、DRwl 2、DRwl 3、DRwl 4、DR 7、DRw 8、DR 9和DRwlO。以前对这些特异性进行分型的方法包括血清学、常规RFLP、PCR-寡核苷酸分型和用于对纯合子个体进行分型的PCR-RFLP方法。在本文报道的方法中，DNA用一组组特异性引物扩增，然后用许多内切核酸酶限制。选择了六对组特异性引物，以避免与其他DRB等位基因（包括DRB 3、DRB 4和DRB 5等位基因）交叉扩增，并在均匀温度（61 ℃）下退火。选择核酸内切酶以产生易于识别的条带的独特模式，从而在杂合子以及纯合子个体中明确分配HLA-DR属型。该技术包括两个步骤：1）用六对不同的引物扩增DNA，其中DR 1、DR 4和DRw 10类型可以一次分配，和2）扩增DNA的内切酶酶切以分配DR 7、DRw 8、DR 9、DRw 11、DRw 12、DRw 13、DRw 14、DRw 15、DRw 16、DRw 17和DRw 18类型。携带迄今为止发表的所有等位基因的任何组合的个体都可以通过这种方法进行分型。讨论了该方法的简便性、低成本和可靠性。
We have described a practical and inexpensive method whereby any individual can be typed and assigned to any of the 14 generic HLA‐DR types: DR1, DRwl5, DRwl6, DRwl7, DRwl8, DR4, DRwll, DRwl2, DRwl3, DRwl4, DR7, DRw8, DR9 and DRw10. Previous methods to type these specificities include ‐ among others ‐ serology, conventional RFLP, PCR‐oligonucleotide typing, and a PCR‐RFLP method useful for typing homozygous individuals. In the method reported here, DNA is amplified with a set of group‐specific primers and then restricted with a number of endonucleases. Six group‐specific pairs of primers have been chosen to avoid cross‐amplification with other DRB alleles, including DRB3, DRB4 and DRB5 alleles, and to anneal at uniform temperature: 61deg;C. Endonucleases were chosen to generate unique patterns of easily recognizable bands that led to unequivocal assignments of HLA‐DR generic types in heterozygous as well as homozygous individuals. This technique involves two steps: 1) Amplification of DNA with six different pairs of primers where DR1, DR4 and DRw10 types can be assigned at once, and 2) Endonuclease digests of amplified DNA to assign DR7, DRw8, DR9, DRw11, DRw12, DRw13, DRw14, DRw15, DRw16, DRw17 and DRw18 types. Individuals carrying any combination of all alleles published so far can be typed by this method. The ease, low cost and reliability of this method are discussed.