Angelini G, Bugawan T L, Delfino L, Erlich H A, Ferrara G B
Istituto Nazionale per la Ricerca sul Cancro, Genova, Italy.
Hum Immunol. 1989 Nov;26(3):169-77. doi: 10.1016/0198-8859(89)90036-0.
HLA-DP genotyping was performed using dot-blot analysis with synthetic oligonucleotide probes. Fourteen probes were designed based on the known sequence variations in the polymorphic segments of the DP beta second exon. Each probe was tested against genomic DNA amplified by the polymerase chain reaction, using DP beta-specific primers. A total of 45 HLA homozygous B-cell lines, selected from the Tenth International Histocompatability Workshop and pretyped for the known DP omega specificities, were analyzed. Different hybridization patterns were found for each DP omega specificity. The oligonucleotide hybridization performed on DP omega-negative B-cell lines gave a pattern distinct from those of known DP omega specificities, indicating the presence of novel DP allelic sequences. The use of sequence-specific oligonucleotides combined with DNA amplification allows a simple and reliable genotyping of DP antigens.
采用合成寡核苷酸探针斑点杂交分析进行HLA - DP基因分型。根据DPβ第二外显子多态性片段的已知序列变异设计了14种探针。使用DPβ特异性引物，针对通过聚合酶链反应扩增的基因组DNA对每种探针进行检测。共分析了45个从第十届国际组织相容性研讨会选取并预先分型为已知DPω特异性的HLA纯合B细胞系。发现每种DPω特异性都有不同的杂交模式。在DPω阴性B细胞系上进行的寡核苷酸杂交产生了与已知DPω特异性不同的模式，表明存在新的DP等位基因序列。序列特异性寡核苷酸与DNA扩增相结合的方法能够对DP抗原进行简单可靠的基因分型。
