毛细管电泳-单链构象多态性分析检测K-ras基因突变

K-ras Gene Mutation Detection by Single-Strand Conformation Polymorphism with Capillary Electrophoresis

K ras癌基因的点突变在结直肠癌的发生起重要作用。以异丙醇为聚合反应链转移剂,水相法合成 特性粘度为0.70×10-3m3/kg,分子量为6.5×104的低粘度短链线性聚丙烯酰胺。以6%线性聚丙烯酰胺为 筛分介质,分离温度27℃,分离电压9kV为电泳条件,建立了检测K ras基因突变的毛细管电泳 单链构象多 态性方法。利用该方法检测36例结直肠癌患者肿瘤组织,发现12例K ras基因突变。结果表明:该方法具有 快速、高灵敏的优点,为大规模进行结直肠肿瘤的早期诊断提供了可靠方法。

Mutation of K-ras gene plays an important role in colorectal cancer tumorigenesis. Short-chain linear polyacrylamide (SLPA) was synthesized using isopropanol as a chain transfer agent. The viscosity coefficient and molecular mass of SLPA were 0.70 x 10(-3) m(3)/kg and 6.5 x 10(4) respectively. A single-strand conformation polymorphism with laser-induced fluorescence capillary electrophoresis method for detection of K-ras gene mutation was developed under the conditions of 6% SLPA, running temperature at 27 degrees C running voltage at 9 kV. By this method, tumor tissues from 36 colorectal cancer patient were screened, and twelve of them were found to be K-ras gene mutation. The results revealed that this method is rapid and high sensitivity and become a promising tool to screen a large number of samples in clinical diagnosis of colorectal cancer.