高效毛细管电泳与聚丙烯酰胺凝胶电泳检测基因点突变的比较

The comparison of detection of gene point mutation by HPCE-SCP and PAGE-SCP analysis

目的比较高效毛细管电泳(HPCE)技术和传统聚丙烯酰胺凝胶电泳(PAGE)检测结肠癌DNA是否存在p53基因点突变的方法。方法分别采用HPCE与PAGE作单链构象多态性(SCP)分析,检测结肠癌肿瘤标本p53基因第7外显子PCR扩增产物,DNA测序检测其准确性。结果利用HPCE技术和PAGE结合银染技术从22例结肠癌标本中分别检测出5例和4例突变,测序证实点突变的确存在。结论HPCE技术较PAGE更为快速、简便和准确,对临床筛查基因点突变有较高的应用价值。

OBJECTIVE To eomparc the method screening point mutation of p53 gene by capillary electrophoresis （CE） and PAGE - SCP. METHODS A simple capillary electrophoresis system for the single strand conformational polymorphism （SCP） analysis of genomic DNA was developed by choosing commercially available capillary and gel buffer. This HPCE - SCP system was applied to analysize 22 PCR products amplified from the exon 7 of the p53 gene of tissue specimens from colon cancer patients. RESULTS Five of samples were found to have mutation,while the results of the SCP analysis by PAGE - silver staining techniques detected only four mutation of samples. Sequencing of PCR products proved that the results were correct. CONCLUSION The HPCE detection is more simple, accurate and less expensive than PAGE - SCP method and could be used in medical research and clinical DNA diagnosis of human cancers and other diseases.