地高辛标记PCR—SSOP法进行HLA—A^＊02等位基因检测

Detection of HLA-A~*02 Alleles Using Polymerase Chain Reaction and Digoxigen-11-2-3 Dideoxyurdinctriphosphate Labeled Oligonucleotide Probes

目的 :建立聚合酶链反应 -序列特异性寡核苷酸探针 (PCR- SSOP)分型方法 ,并分析其实用价值。方法 :采用一对引物扩增所有 HL A- A2特异性抗原 ,用 2 3个探针和扩增产物杂交 ,探针用地高辛系统标记和检测 ,可鉴定 A2的 2 5个等位基因 ,并在中国人群 HL A- A*0 2等位基因的检测中进行应用 ,用 B淋巴母细胞株作阳性对照 ,同时与聚合酶链反应 -序列特异性引物 (PCR- SSP)检测方法作比较。结果 :采用本方法检测 HL A - A 2等位基因的结果与用 PCR- SSP方法检测的结果一致。结论 :实验证明本分型技术具有质控好、特异性强、敏感性高、费用低廉。

Objective:The experiment was designed to establish a method of polymerase chain reaction and sequence specific oligonucleotide probes (PCR SSOP) for identification HLA A2 alleles, and to analyze the practicability of the method. Methods: A PCR SSOP typing method, involving a single PCR amplification in conjunction with 23 digoxigenin labeled oligonucleotide probes, has been employed for the identification of 25 known HLA A *02 alleles. Twenty A2 (+) samples in Shenyang were analyzed with this method and compared with PCR SSP typing method. Results: The finding of our method was same with those of PCR SSP. Conclusion:The method is highly specific and sensitive. The finding of this study has implications for the selection of donor in unrelated bone marrow transplantation and organ transplantation and human genetic studies.