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目的 应用寡核苷酸芯片技术检测胃癌p5 3基因突变的价值。方法 提取胃癌基因组DNA ,以荧光标记引物进行PCR ,扩增p5 3基因特异性片段 ;将修饰合成的探针按微矩阵排列点样于化学处理过的载玻片上 ,制成寡核苷酸芯片 ;将PCR产物与寡核苷酸芯片进行杂交 ,经严格洗片后在激光共聚焦扫描仪上扫描芯片 ,获取图象分析结果。结果 检测 10例胃癌组织标本的p5 3基因突变情况 ,发现其中 7例野生型 ,3例在第 7外显子发生点突变。结论 p5 3基因突变与胃癌发生机制有关 ,同时芯片技术在检测基因突变方面的应用将对胃癌研究的基础理论及临床应用具有重要意义
Objective To evaluate the value of oligonucleotide microarray in detecting the mutation of p5 3 gene in gastric cancer. Methods Genomic DNA of gastric cancer was extracted and PCR was carried out with fluorescently labeled primers to amplify specific fragments of p5 3 gene. The modified synthetic probes were microarrayed on chemically treated glass slides to prepare oligonucleotide chips The PCR product was hybridized with the oligonucleotide chip, and the chip was scanned on the laser confocal scanner after being strictly washed to obtain the image analysis result. Results The mutation of p5 3 gene in 10 specimens of gastric cancer tissues was detected. Among them, 7 cases were wild type and 3 cases were point mutation in exon 7. Conclusions The mutation of p5 3 gene is related to the mechanism of gastric cancer, and the application of chip technology in detecting gene mutation will be of great significance to the basic theory and clinical application of gastric cancer research