DHPLC detection of p53gene mutation in plasma DNA from40malignant pa┐tients

    WANG Hai-Dan，REN Jun，LU Shao-Min，SI Xiao-Ming，PAN Bo-Rong

    Keywords:p53gene;DNA，plasma;chromatography，liquid CLC number:R735 Document code:A

    Abstract: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 of40malignant patients by using Qia-gen Blood Kit and the target DNA fragments were amplified by PCR，the production was analyzed by denaturing high per-formance liquid chromatography(DHPLC)，and the mutation status obtained by DHPLC was proved by DNA sequencing.RESULTS The target fragments were obtained from all samples of40malignant patients and normal tissue DNA，8mutations were detected by DHPLC and confirmed by DNA sequencing from malignant patients，no mutation was found in examples that no mutation detected by DHPLC.CONCLU┐SION Plasma DNA can be amplified in malignant patients，DHPLC is an accurate，rapid and economic way to screen mu-tation in plasma DNA amplified fragments and plasma p53gene mutation detection may be useful for cancer alerting ......