CAJ | 학술논문

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40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,随机送检的8份DHPLC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,DHPLC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p53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.
목적탐토확증악성종류환자혈장DNA적가행성,건립일충쾌속검측혈장DNA돌변적신방법. 방법선취악성종류환자공40례,채용Qiagen column주추제법제취혈장DNA,채용취합매련반응(PCR)확증p53기인외현자7,용변성고효액상색보법(DHPLC)대산물진행돌변분석,병여측서결과비교. 결과 40례악성종류환자혈장제취적DNA균능확증출목적편단,DHPLC검측도유8례돌변,수궤송검적8빈DHPLC미검측도돌변자측서야미발현돌변존재,여DNA직접측서결과상일치. 결론대악성종류환자혈장DNA행PCR확증시가행적,DHPLC가작위일충쾌속,간편화경제적돌변사선방법,운용차방법검측혈장중p53등기인적돌변유망응용우악성종류고위인군적예경화조기진단이급작위예후삼고.
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.