用单链构型多态性快速检测血小板膜糖蛋白Ⅱb和Ⅲa基因变异

为探讨快速检测血小板膜的糖蛋白和Ⅲａ基因点突变的方法，采用单链构型多态性分析研究正常人和血小板无力症患者血小板膜ＧＰⅢｂ，Ⅲａ基因结构。外周血ＤＮＡ提取后，用聚合酶链反主尖扩增各外显子，ＰＣＲ产物变性后在微型聚丙烯酰胺凝胶上分离，通过银染色对迁移率异常的ＰＣＲ产物进行核苷酸序列分析。

Rapid detection of platelet glycoprotein llb , IIIa genevariety by single strand confor''iiation Polymorphismanalysis

lanzmann thrombasthenia (GT) is the most com-mon inherited disorder of platelet defective function.Most of the molecular defects of GT identified in recentyears have been caused by point mutation in the plateletglycoprotein (GP) IIb , IIIa genes. Trying to rapidly de-tect point mutation (or subtle variety) in GPllb, IIIagenes, we have used single strand conformation poly-morphism analysis (SSCPA) and studied the gene struc-ture of platelet GPIIb, IIIa from normal controls andpatients with Glanzmann thrombasthenia. After DNApreparation from peripheral bloed . all the exons of GPI-Ib, IIIa were amplified by PCR. Denaturated PCRproducts were separated on mini polyacrylamide gel andstained by silver staining method in the Phast System.Only the PCR products with different migration on thegel were sequenced. 2 GT patients with point mutationsand 1 GT patient with 16 bp deletion were found in ourstudy. At the same time, we determined 3 polymor-phisms in GPllb gene. We conclude that SSCPA is arapid, sensitive and effective methed for detectingplatelet IIb, IIIa gene variety.