试剂盒配合琼脂糖EB染色法和PAGE银染法分析线粒体DNA A1555G突变的比较研究

目的 通过比较试剂盒配合琼脂糖EB染色法和PAGE银染法分析线粒体DNA A1555G突变的不同特点,探索应用标准试剂盒和PAGE银染方法检测线粒体DNA A1555G突变的快速筛查和诊断方法.方法 采用线粒体DNA A1555G突变诊断试剂盒配合PAGE胶电泳银染法检测线粒体DNA 1555位点正常者229例,A＞G突变阳性者17例,验证此试剂盒配合PAGE银染法检测的有效性和准确性.结果 线粒体DNA A1555G突变诊断试剂盒配合PAGE胶电泳银染法检测结果与酶切EB染色和测序结果完全吻合.结论 线粒体DNA A1555G突变诊断试剂盒配合PAGE胶电泳银染法和试剂盒配合EB检测法较前者更为灵敏和清晰,在分析线粒体DNA A1555G突变方面具有简单、价廉、结果直观的特点,适合在中国用于进行此突变的大规模筛查或预防性检查.

Mitochondrial DNA A1555G Mutation Analyzed by Testing Kit with PAGE Silver Staining and Ethidium Bromide Staining Methods

Objective To establish a new diagnostic method of mtDNA A1555G mutation using testing kit with PAGE silver staining and to evaluate the feasibility of the method as a fast diagnostic technique. Methods 229 samples including 17 positive cases carrying mtDNA A1555G mutation were tested by mtDNA A1555G mutation testing kit with PAGE silver staining methods and the results of this method were compared with the results of ethidium bromide staining and direct sequencing, Results 17 positive samples were found to carry the mtDNA A1555G mutation by the method of the testing kit and PAGE silver staining. The reslts were all confirmed by ethidium bromide staining and sequence analysis. Conclusion mtDNA A1555G mutation testing kit and PAGE silver staining methods are more sensitive than ethidium bromide staining. They are convenient, inexpensive, and suitable in China for large scale screening and preventive testing.