HLA-C high resolution typing: analysis of exons 2 and 3 by sequence based typing and detection of polymorphisms in exons 1-5 by sequence specific primers

被引:21
作者
Delfino, L [1 ]
Morabito, A [1 ]
Longo, A [1 ]
Ferrara, GB [1 ]
机构
[1] IST, Natl Canc Inst, Adv Biotechnol Ctr, Genoa, Italy
来源
TISSUE ANTIGENS | 1998年 / 52卷 / 03期
关键词
HLA-C; PCR-SBT; PCR-SSP; serological typing;
D O I
10.1111/j.1399-0039.1998.tb03040.x
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
HLA-C high resolution sequence based typing developed in this study involves a unique DNA amplification encompassing exon 1 to intron 3 and four fluorescent sequencing reactions covering exon 2 and 3. Both dye primer and dye terminator sequencing techniques were performed and results compared. This approach allowed the identification of all of the 50 HLA-C allelic variants so far described, except for two allele pairs that are distinguished by non-coding nucleotide changes (Cw*12021=12022, Cw*15051 = 15052) and three allele pairs (Cw*0701 = 0706, Cw*1701 = 1702 and Cw*1801 = 1802) that share the same nucleotide sequence in exon 2 and 3. For complete subtyping of these allelic variants, an amplification based on sequence specific primers (PCR-SSP) was used. No ambiguous heterozygous combinations of alleles were detected in our panel so far. HLA-C typing data obtained by this method were compared with data from serological and low resolution PCR-SSP typing, which had been performed previously on the samples sequenced.
引用
收藏
页码:251 / 259
页数:9
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