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目的建立ras基因寡核苷酸芯片对胰腺癌基因突变的检测系统。评估16例胰腺癌组织k-ras基因12、13、61位密码子变突检测。方法采用双重或单独不对称PCR扩增标本中的目的DNA。扩增产物加杂交液后与芯片进行杂交、清洗、扫描。结果16例胰腺癌组织中k-ras为75%,突变都发生在12密码子,k-ras以12密码子第2位核苷酸突变发生率高(8/12)。结论k-ras基因芯片系统具有高度的灵敏性和准确性、快速简便、自动化程度高等优点,可同时检测胰腺癌k-ras多个突变位点基因,有利于临床应用。
Objective To establish a ras gene oligonucleotide microarray for detection of pancreatic cancer gene mutation. To evaluate the mutation of codon 12, 13 and 61 of k-ras gene in 16 cases of pancreatic cancer. Methods Double or single asymmetric PCR was used to amplify the DNA of interest. Amplification products plus hybridization liquid hybridization with the chip, cleaning, scanning. Results The k-ras was 75% in 16 cases of pancreatic cancer, the mutation occurred at 12 codons, and the incidence of k-ras mutation at nucleotide 12 of 12 codons was high (8/12). Conclusion The k-ras gene chip system has the advantages of high sensitivity and accuracy, quick and easy, and high degree of automation. It can simultaneously detect multiple k-ras gene in pancreatic cancer and is helpful for clinical application.