用变性高效液相色谱法检测遗传性因子XⅢA链基因突变  

DETECTION OF GENE MUTATION IN FXIIIA CHAIN GENE BY DHPLC

作　　者：王巍[1] 陈晓丹[2] 蔡望伟[3] 朱燕[4] 刘国勋[5] 程爽[5] 

机构地区：[1]广东医学院检验学院临床血液检验学教研室,湛江524023 [2]海南医学院附属医院检验科,海口570102 [3]海南医学院生物化学教研室,海口571101 [4]海南医学院附属医院儿科,海口570102 [5]广东医学院附属医院血液研究室,湛江524023

出　　处：《海南医学院学报》2004年第6期378-380,共3页Journal of Hainan Medical University

基　　金：国家自然科学基金(30060037)教育部科学技术研究重点项目(No.03147)

摘　　要：目的:观察变性高效液相色谱(DHPLC)法检测凝血因子XIIIA链基因突变的实用价值。方法:用聚合酶链反应(PCR)扩增正常样品和有XIIIA基因603-606delAAGA突变患者的XIIIA链基因外显子5;用单链构象多态性(SSCP)及DHPLC检测分析扩增的外显子5。结果:SSCP显示患者有异常的区带,患者的DHPLC结果出现异常洗脱峰,与SSCP的结果一致。结果表明DHPLC能检测FXIII因子A链基因突变。结论:DHPLC技术是一种筛查基因突变的快速、简单、可靠的方法。Objective: To observe the value of detecting mutation in coagulation factor XIIIA chain gene by denatured high performance liquid chromatography (DHPLC). Methods: Both Exon 5 of FXIIIA gene of normal sample and the patient with 603 -606delAAGA mutation in FXIIIA gene were amplified by polymerase chain reaction. SSCP and DHPXC were used to analyze the amplified fragment of exon 5. Results: Band-shift was found in the SSCP in the patient. An abnormal peak, corresponding to the SSCP result, was found in exon 5 of the patient by DHPLC. The results indicated that DHPLC can detect the mutation of FXIIIA gene. Conclusions: DHPLC technique is a rapid, simple and reliable method for screening gene mutation.

关 键 词：基因突变 DHPLC 患者 SSCP 变性高效液相色谱法 扩增 遗传性 外显子 洗脱 单链构象多态性 

分 类 号：R735[医药卫生—肿瘤] R739.63[医药卫生—临床医学]