聚合酶链反应顺序特异性寡聚核苷酸探针反向杂交法在HLA-DRB1分型中的应用

目的:探讨聚合酶链反应顺序特异性寡聚核苷酸探针(polymerasechainreaction-sequencespe-cificoligonucleotideprobes,PCR-SSOP)反向杂交法应用于人类白细胞抗原(humanleukocyteantigen,HLA)-DRB1配型的可行性。方法:随机选取25例已行聚合酶链反应顺序特异性引物(polymerasechainreac-tion-sequencespecificprimers,PCR-SSP)法HLA-DRB1配型的肾移植受者的血标本,采用PCR-SSOP反向杂交法再次配型,并与PCR-SSP法的结果比较。结果:两法的一致性达92%。2例不一致的患者经再次PCR-SSOP反向杂交法配型后,其中1例与PCR-SSP结果一致,PCR-SSOP反向杂交法的平均操作时间为4小时30分。结论:PCR-SSOP反向杂交法是一种能以中等分辨度进行等位基因分型的方法,操作较简单,结果解读客观,适用于临床肾移植配型。

The use of polymerase chain reaction-sequence specific oligonucleotide probes reverse hybridization in HLA-DRB1 typing

Objective:To explore the use of polymerase chain reaction sequence specific oligonucleotide probes (PCR SSOP) reverse hybridization in human leukocyte antigen (HLA) DRB1 typing. Methods:Twenty five blood samples from patients who had performed HLA DRB1 typing by polymerase chain reaction sequence specific primers (PCR SSP) method were performed typing again with PCR SSOP reverse hybridization method. Results:The concordance rate between the two methods was 92%,One of the two cases with different results got identical result after repeated examination. The average time cost for PCR SSOP was 4.5 hours. Conclusion:PCR SSOP reverse hybridization can give an intermediate allelic level resolution.The manipulation is simple and easy to learn. The interpretation of the results is objective and the method is suitable for clinical renal transplantation typing.