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

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.