应用PCR-SSP和SSOP对HLA-I类进行基因分型

• 作 者: 傅茜 曾文涛 邓素雄 陈连周 王长希
• 机 构: 中山大学附属第一医院,中山大学附属第一医院,中山大学附属第一医院,中山大学附属第一医院,中山大学附属第一医院 510080广州市,510080广州市,510080广州市,510080广州市,51008
• 出 处: 实用医学杂志
• 发表日期: 2004年11期
• 关键词: 基因 MHCⅠ类 基因型 PCR-SSP PCR-SSOP 器官移植

目的 :探讨序列特异性引物 -聚合酶链反应 (polymerasechainreaction sequence specificprimer ,PCR SSP)方法和多聚酶链反应序列特异性寡聚核苷酸探针 (PCR sequence specificoligonucleotideprobes ,PCR SSOP)技术在HLA I类基因分型的应用价值。方法 :研究样本 3 0份 ,为等待肾移植供、受者外周血。采用PCR SSOP反向杂交技术对HLA I进行基因分型 ,并将两种方法分型结果进行比较研究。结果 :所有样本进行HLA I分型均获得成功 ,分型结果PCR SSOP与PCR SSP相符率为 93 3 % ,SSOP分辨率较SSP高。结论 :PCR SSP方法快捷简便 ,适用于少量标本。PCR SSOP分型方法准确率与SSP接近 ,分辨率高 ,虽检测单份标本耗时较长 ,却可同时检测大批量的样本 ,适应临床器官移植组织配型短时高效的需要

OBJECTIVE: To investigate the effect of HLA class I genotyping by PCR SSP and PCR SSOP (polymerase chain reaction sequence specific polymerase chain reaction) Type of application value. Methods: 30 samples were studied, for the purpose of waiting for kidney transplantation for peripheral blood of recipients. The PCR SSOP reverse hybridization was used to genotype HLA I, and the results of two methods were compared. Results: All the samples were successfully genotyped by HLA I. The typing rate of PCR SSOP was the same as PCR SSP 93 3%, and the resolution of SSOP was higher than that of SSP. Conclusion: PCR SSP method is quick and easy, suitable for a small amount of specimens. The accuracy of PCR SSOP typing method is close to that of SSP, and its resolution is high. Although it takes a long time to detect a single specimen, it can simultaneously detect large quantities of samples to meet the short-term and high-efficiency requirements of clinical organ transplantation