HLA-DR快速基因分型PCR-SSP方法的建立  

A rapid method for HLA DR typing with PCR SSP

作　　者：曹孟德[1,2] 秦东春[1,2] 郭小兵 苏堤[1,2] 王东升[1,2] 苏天水 燕桂香[1,2] 

机构地区：[1]河南医科大学微生物与免疫学教研室 [2]河南医科大学第一附属医院检验科

出　　处：《河南医科大学学报》1999年第1期91-93,共3页Journal of Henan Medical University

基　　金：河南省科技攻关资助项目

摘　　要：目的：探讨应用于器官移植配型及疾病相关性分析的方法及基本数据，建立一种高分辨率、高特异性、简单、快速、方便的ＨＬＡＤＲ基因分析方法。方法：利用ＤＲ１～ＤＲ１８序列特异性引物及１对内对照引物，采用序列特异性引物聚合酶链式反应（ｐｏｌｙｍｅｒａｓｅｃｈａｉｎｒｅａｃｔｉｏｎｓｅｑｕｅｎｃｅｓｐｅｃｉｆｉｃｐｒｉｍｅｒｓ，ＰＣＰＳＳＰ）技术对２２份国际标准细胞株ＤＮＡ及２０例未知ＤＲ型别的临床血液标本进行ＨＬＡＤＲ基因分析，扩增条件为：变性９４℃，６０ｓ；退火６５℃，６０ｓ；延伸７２℃，６０ｓ。２５个循环后，７２℃保温７ｍｉｎ。结果：对各个标准ＤＮＡ的分型结果显示，准确率及重复率均为１００％，无假阳性及假阴性。结论：该方法快速、简便、灵敏、重复性好，结果易于判断，适合临床器官移植组织配型、疾病相关性分析、法医鉴定及人类学研究等。Aim: To discuss the establishing of a rapid method for HLA DR typing. Method: Using polymerase chain reaction with sequence specific primers (PCR SSP) that was carried out in 22 standard cell lines DNA and 20 individuals, twenty separate PCR reactions were performed on each sample. The amplification was accomplished through 25 cycles consisting of denaturation at 94 ℃ for 60 s, annealing at 65 ℃ for 60 s and extension at 72 ℃ for 60 s, and incubation at 72 ℃ for 7 min after 25 cycles. Results: Reproducible rate of the PCR SSP typing results was 100%. All typing tests were successful. False positive or false negative typing results were not obtained. Conclusion: Genotyping by PCR SSP is a rapid and accurate technique and suitable for clinical practice.

关 键 词：HAL-DR抗原 基因分型 器官移植 PCR-SSP 

分 类 号：R617[医药卫生—外科学] R392.4[医药卫生—临床医学]