变性高效液相色谱法对恶性肿瘤患者血浆DNA p53突变的检测

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40例,提取血浆DNA,采用聚合酶链反应(PCR)扩增p53基因外显子7,用变性高效液相色谱法(DHPLC)对产物进行突变分析,并与测序结果比较。结果:40例恶性肿瘤患者血浆提取的DNA均能扩增出目的片断,DHPLC检测到有8例突变,与DNA直接测序结果一致。结论:对恶性肿瘤患者血浆DNA扩增是可行的,DHPLC可作为一种快速简便的突变检测方法。

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

Objective：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： Extraction DNA from plasma of 40 malignant patients by using QIAGen Blood Kit and the target DNAfragments were amplified by PCR,the production were analyzed by denaturing high performance liquid chromatography（DHPLC）,the mutation status obtained by DHPLC was proved by DNA sequenced.Results：The target fragments were obtained from all samples of 40 malignant patients,8 mutation of the fragment were detected by DHPLC and confirmed by DNA sequencing.Conclusion：Plasma DNA can be amplified in malignant patients,DHPLC is a accurate,rapid and economics way to screen mutation in plasma DNA amplified fragment.