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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 .
Objective To explore the feasibility of amplifying plasma DNA in patients with malignant tumors and establish a new method for rapid detection of plasma DNA mutations. Methods A total of 40 patients with malignant tumors were selected. Plasma DNA was extracted using Qiagen column method and polymerase chain was used. The exon 7 of p53 gene was amplified by PCR (PCR), and the product was analyzed by denaturing high performance liquid chromatography (DHPL C). The results were compared with the sequencing results. Results The DNA extracted from plasma of 40 patients with malignant tumor was able to The target fragment was amplified, and 8 mutations were detected by DHPL C. No mutations were detected in the 8 randomly sampled DHPL C. No sequencing was found. The results were consistent with the direct DNA sequencing results. Conclusion Plasma DNA from patients with malignant tumors PCR amplification is feasible, and DHPL C can be used as a rapid, simple and economical mutation screening method. Detection of p53 gene mutations in plasma may be used for early warning and early diagnosis of high-risk cancers and as a method of detection. Prognosis reference