变性高效液相色谱检测恶性肿瘤患者血浆 DNA p53突变(英文)

DHPLC detection of p53 gene mutation in plasma DNA from 40 malignant patients

目的 探讨扩增恶性肿瘤患者血浆 DNA的可行性 ,建立一种快速检测血浆 DNA突变的新方法 .方法 选取恶性肿瘤患者共 4 0例 ,采用 Qiagen column柱抽提法提取血浆DNA,采用聚合酶链反应 (PCR)扩增 p5 3基因外显子 7,用变性高效液相色谱法 (DHPL C)对产物进行突变分析 ,并与测序结果比较 .结果 4 0例恶性肿瘤患者血浆提取的 DNA均能扩增出目的片断 ,DHPL C检测到有 8例突变 ,随机送检的 8份 DHPL C未检测到突变者测序也未发现突变存在 ,与 DNA直接测序结果相一致 .结论 对恶性肿瘤患者血浆 DNA行PCR扩增是可行的 ,DHPL C可作为一种快速 ,简便和经济的突变筛选方法 ,运用此方法检测血浆中 p5 3等基因的突变有望应用于恶性肿瘤高危人群的预警和早期诊断以及作为预后参考 .

AIM To evaluate the presence of tumor DNA in plasma from malignant patients and establish a new approach to detect gene mutation in plasma DNA. METHODS DNA extraction from plasma of 40 malignant patients by using Qiagen Blood Kit and the target DNA fragments were amplified by PCR, the production was analyzed by denaturing high performance liquid chromatography (DHPLC), and the mutation status obtained by DHPLC was proved by DNA sequencing. RESULTS The target fragments were obtained from all samples of 40 malignant patients and normal tissue DNA, 8 mutations were detected by DHPLC and confirmed by DNA sequencing from malignant patients, no mutation was found in examples that no mutation detected by DHPLC. CONCLUSION Plasma DNA can be amplified in malignant patients, DHPLC is an accurate, rapid and economic way to screen mutation in plasma DNA amplified fragments and plasma p53 gene mutation detection may be useful for cancer alerting, early diagnosis or prognosis prediction.